Structured Review

Bethyl total human igd
<t>IgD</t> Ligation by Antigen Elicits Basophil Activation (A) <t>ELISA</t> of serum IgD from WT Balb/c (n=29), WT C57BL/6 (n=15), Ighd −/− Balb/c (n=20) or Ighd −/− C57BL/6 (n=10) mice. (B) Left: FCM of CD49b and IgE on basophils (black gate) from circulating CD45 + NTNB cells of a WT Balb/c mouse. NTNB, non-T non-B. Right: FCM of IgD on circulating basophils (blue open profile) from a WT Balb/c mouse. Ctrl, isotype-matched control (gray solid profile). (C) FCM of IgD + basophils from peripheral blood, spleen, bone marrow (BM), lung or mesenteric lymph nodes (MLNs) of WT Balb/c mice (n=5). (D) Imaging FCM of IgD, FcεRI and CD49b from a representative viable Ghost Dye Violet 510 (GV510) − splenic basophil of a WT Balb/c mouse. Scale bar, 5 μm. (E) FCM quantification of FcεRI + CD49b + basophils from BM, blood, spleen or lung CD45 + NTNB cells of WT Balb/c (n=10) or Ighd −/− (n=10) mice. (F) FCM of Il4 -driven GFP expression in splenic or lung FcεRI + CD49b + basophils from WT Balb/c Il4 GFP (n=5) or Il4 GFP Ighd −/− (n=5) mice. MFI, mean fluorescence intensity. (G) ELISA of IL-4 and IL-13 from serum of WT Balb/c or Ighd −/− mice (n=10). (H) Schematics of i.v. reconstitution μMT or Rag2 −/− mice with NP-reactive IgD (NP-IgD) followed by i.p. injection of anti-IgD or i.n. inoculation of NP-OVA. (I) FCM of IgD on splenic FcεRI + CD49b + basophils from a μMT mouse before (ctrl) or after reconstitution as in (H). PBS, control phosphate buffer solution (PBS). (J, K) FCM quantification of total, CD200R3 + or IL-4 + basophils from the spleen of μMT C57BL/6 mice (n=10) (J) or the lungs of IgD-deficient Rag2 −/− (n=10) C57BL/6 mice (K) treated as in (H). (L) FCM quantification of total, IgD + , IgE + or IL-4 + basophils from the lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice 5 d following i.n. exposure to PBS or recombinant TSLP. (M) ELISA of serum IgD from Balb/c mice following i.v. injection of control empty or TSLP-encoding plasmids (n=10). .
Total Human Igd, supplied by Bethyl, used in various techniques. Bioz Stars score: 92/100, based on 568 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/total human igd/product/Bethyl
Average 92 stars, based on 568 article reviews
Price from $9.99 to $1999.99
total human igd - by Bioz Stars, 2020-08
92/100 stars

Images

1) Product Images from "Secreted IgD Amplifies Humoral T Helper-2 Responses by Binding Basophils Via Galectin-9 and CD44"

Article Title: Secreted IgD Amplifies Humoral T Helper-2 Responses by Binding Basophils Via Galectin-9 and CD44

Journal: Immunity

doi: 10.1016/j.immuni.2018.08.013

IgD Ligation by Antigen Elicits Basophil Activation (A) ELISA of serum IgD from WT Balb/c (n=29), WT C57BL/6 (n=15), Ighd −/− Balb/c (n=20) or Ighd −/− C57BL/6 (n=10) mice. (B) Left: FCM of CD49b and IgE on basophils (black gate) from circulating CD45 + NTNB cells of a WT Balb/c mouse. NTNB, non-T non-B. Right: FCM of IgD on circulating basophils (blue open profile) from a WT Balb/c mouse. Ctrl, isotype-matched control (gray solid profile). (C) FCM of IgD + basophils from peripheral blood, spleen, bone marrow (BM), lung or mesenteric lymph nodes (MLNs) of WT Balb/c mice (n=5). (D) Imaging FCM of IgD, FcεRI and CD49b from a representative viable Ghost Dye Violet 510 (GV510) − splenic basophil of a WT Balb/c mouse. Scale bar, 5 μm. (E) FCM quantification of FcεRI + CD49b + basophils from BM, blood, spleen or lung CD45 + NTNB cells of WT Balb/c (n=10) or Ighd −/− (n=10) mice. (F) FCM of Il4 -driven GFP expression in splenic or lung FcεRI + CD49b + basophils from WT Balb/c Il4 GFP (n=5) or Il4 GFP Ighd −/− (n=5) mice. MFI, mean fluorescence intensity. (G) ELISA of IL-4 and IL-13 from serum of WT Balb/c or Ighd −/− mice (n=10). (H) Schematics of i.v. reconstitution μMT or Rag2 −/− mice with NP-reactive IgD (NP-IgD) followed by i.p. injection of anti-IgD or i.n. inoculation of NP-OVA. (I) FCM of IgD on splenic FcεRI + CD49b + basophils from a μMT mouse before (ctrl) or after reconstitution as in (H). PBS, control phosphate buffer solution (PBS). (J, K) FCM quantification of total, CD200R3 + or IL-4 + basophils from the spleen of μMT C57BL/6 mice (n=10) (J) or the lungs of IgD-deficient Rag2 −/− (n=10) C57BL/6 mice (K) treated as in (H). (L) FCM quantification of total, IgD + , IgE + or IL-4 + basophils from the lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice 5 d following i.n. exposure to PBS or recombinant TSLP. (M) ELISA of serum IgD from Balb/c mice following i.v. injection of control empty or TSLP-encoding plasmids (n=10). .
Figure Legend Snippet: IgD Ligation by Antigen Elicits Basophil Activation (A) ELISA of serum IgD from WT Balb/c (n=29), WT C57BL/6 (n=15), Ighd −/− Balb/c (n=20) or Ighd −/− C57BL/6 (n=10) mice. (B) Left: FCM of CD49b and IgE on basophils (black gate) from circulating CD45 + NTNB cells of a WT Balb/c mouse. NTNB, non-T non-B. Right: FCM of IgD on circulating basophils (blue open profile) from a WT Balb/c mouse. Ctrl, isotype-matched control (gray solid profile). (C) FCM of IgD + basophils from peripheral blood, spleen, bone marrow (BM), lung or mesenteric lymph nodes (MLNs) of WT Balb/c mice (n=5). (D) Imaging FCM of IgD, FcεRI and CD49b from a representative viable Ghost Dye Violet 510 (GV510) − splenic basophil of a WT Balb/c mouse. Scale bar, 5 μm. (E) FCM quantification of FcεRI + CD49b + basophils from BM, blood, spleen or lung CD45 + NTNB cells of WT Balb/c (n=10) or Ighd −/− (n=10) mice. (F) FCM of Il4 -driven GFP expression in splenic or lung FcεRI + CD49b + basophils from WT Balb/c Il4 GFP (n=5) or Il4 GFP Ighd −/− (n=5) mice. MFI, mean fluorescence intensity. (G) ELISA of IL-4 and IL-13 from serum of WT Balb/c or Ighd −/− mice (n=10). (H) Schematics of i.v. reconstitution μMT or Rag2 −/− mice with NP-reactive IgD (NP-IgD) followed by i.p. injection of anti-IgD or i.n. inoculation of NP-OVA. (I) FCM of IgD on splenic FcεRI + CD49b + basophils from a μMT mouse before (ctrl) or after reconstitution as in (H). PBS, control phosphate buffer solution (PBS). (J, K) FCM quantification of total, CD200R3 + or IL-4 + basophils from the spleen of μMT C57BL/6 mice (n=10) (J) or the lungs of IgD-deficient Rag2 −/− (n=10) C57BL/6 mice (K) treated as in (H). (L) FCM quantification of total, IgD + , IgE + or IL-4 + basophils from the lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice 5 d following i.n. exposure to PBS or recombinant TSLP. (M) ELISA of serum IgD from Balb/c mice following i.v. injection of control empty or TSLP-encoding plasmids (n=10). .

Techniques Used: Ligation, Activation Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay, Imaging, Expressing, Fluorescence, Injection, Recombinant

Basophil-Bound IgD Interacts with Galectin-9 and Its Ligation Induces Th2 Cell-Associated Cytokine Expression But Inhibits Cytoskeleton Remodeling (A) Schematics of i.p. anti-IgD treated mice i.v. reconstituted or not with secreted IgD. (B-C) ELISA of serum IgG1, IgE (B), IL-4 and IL-13 (C) from WT Balb/c (n=10) or Ighd −/− (n=10) mice in the presence or absence of i.v. reconstitution with secreted IgD (NP-IgD), followed by i.p. injection of anti-IgD. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to onset of anti-IgD treatment (day 0). (D) IFA of human tonsillar tissue stained for IgD (green), IgM (red), cytokeratin (purple) and nuclei (blue). EP, epithelium; FO, follicle; PC, plasma cell. Original magnification, ×20 (top) or ×63 (bottom). Scale bars, 50 μm (top) and 5 μm (bottom). (E) ELISA of IgD specific to α-s-casein, β-lactoglobulin (β-LGB) or α-lactalbumin (α-LAL) from plasma of FPIES children with (n=5) or without (n=5) dietary milk restrictions. (F) FCM of IgD (red open profile) bound to human tonsillar or circulating CD123 + FcεRI + basophils (red gate). Gray open profiles, control isotype-matched antibody with irrelevant binding activity. (G) Microarray analysis of genes expressed by human basophils upon IgD cross-linking. The Volcano plot represents genes differentially expressed by basophils treated with anti-IgD or a F(ab’)2 control antibody (ctrl). Red and blue dots, up-regulated and down-regulated genes, respectively; FC, fold change. (H) Heat map of coordinated gene sets identified by gene set enrichment analysis in human basophils treated as in (G). NES (normalized enrichment score) indicates correlation between individual gene sets. Positive correlation, NES > 0 (yellow gradient); negative correlation, NES
Figure Legend Snippet: Basophil-Bound IgD Interacts with Galectin-9 and Its Ligation Induces Th2 Cell-Associated Cytokine Expression But Inhibits Cytoskeleton Remodeling (A) Schematics of i.p. anti-IgD treated mice i.v. reconstituted or not with secreted IgD. (B-C) ELISA of serum IgG1, IgE (B), IL-4 and IL-13 (C) from WT Balb/c (n=10) or Ighd −/− (n=10) mice in the presence or absence of i.v. reconstitution with secreted IgD (NP-IgD), followed by i.p. injection of anti-IgD. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to onset of anti-IgD treatment (day 0). (D) IFA of human tonsillar tissue stained for IgD (green), IgM (red), cytokeratin (purple) and nuclei (blue). EP, epithelium; FO, follicle; PC, plasma cell. Original magnification, ×20 (top) or ×63 (bottom). Scale bars, 50 μm (top) and 5 μm (bottom). (E) ELISA of IgD specific to α-s-casein, β-lactoglobulin (β-LGB) or α-lactalbumin (α-LAL) from plasma of FPIES children with (n=5) or without (n=5) dietary milk restrictions. (F) FCM of IgD (red open profile) bound to human tonsillar or circulating CD123 + FcεRI + basophils (red gate). Gray open profiles, control isotype-matched antibody with irrelevant binding activity. (G) Microarray analysis of genes expressed by human basophils upon IgD cross-linking. The Volcano plot represents genes differentially expressed by basophils treated with anti-IgD or a F(ab’)2 control antibody (ctrl). Red and blue dots, up-regulated and down-regulated genes, respectively; FC, fold change. (H) Heat map of coordinated gene sets identified by gene set enrichment analysis in human basophils treated as in (G). NES (normalized enrichment score) indicates correlation between individual gene sets. Positive correlation, NES > 0 (yellow gradient); negative correlation, NES

Techniques Used: Ligation, Expressing, Mouse Assay, Enzyme-linked Immunosorbent Assay, Injection, Concentration Assay, Immunofluorescence, Staining, Binding Assay, Activity Assay, Microarray

IgD Binds to Basophils through Galectin-9 and CD44 (A) IB of galectin-9 following IP of human IgD and galectin-9 protein mix with control (ctrl) or anti-IgD antibodies. Prior to IP, the protein mix was supplemented with control PBS, glucose or lactose. (B) FCM of human IgD on human KU812 cells cultured for 30 min with control medium alone (ctrl), IgD or an IgD-galectin-9 complex formed by pre-incubating IgD with galectin-9 for 10 min. (C) FCM of human IgD or galectin-9 on KU812 cells cultured with IgD-galectin-9 in the presence of medium alone (ctrl), glucose or lactose for 30 min. (D) FCM of human IgD or CD44 on KU812 cells treated with scrambled (ctrl) or CD44-targeting small interfering RNA (siRNA) and later incubated with or without (ctrl) IgD-galectin-9. MFI, mean fluorescence intensity. (E) Confocal imaging of human basophils stained for IgD (blue), galectin-9 (green) and CD44 (red). Scale bar, 0.5 μm. (F) FCM of IgD on human basophils incubated with or without (ctrl) IgD-galectin-9 for 30 min. (G) ELISA of IL-4 from human basophils incubated with medium alone (ctrl) and with or without the IgD-galectin-9 complex in the presence or absence of IgD cross-linking by anti-IgD for 18 h. (H, I) FCM of IgD + basophils from the spleen or lung of WT C57BL/6, Lgals9 −/− (H) or Cd44 −/− (I) mice. (J, K) ELISA of serum IgG1 and IgE to NP from WT Balb/c (n=10), Lgals9 −/− (n=8) (J) or Cd44 −/− (n=10) (K) mice following s.c. immunization with NP-OVA and papain. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of anti-IgD treatment (day 0). (L) ELISA of serum total IgG1 and IgE from WT C57BL/6 controls (n=10) and Lgals9 −/− mice (n=8) following i.p. injection of anti-IgD. .
Figure Legend Snippet: IgD Binds to Basophils through Galectin-9 and CD44 (A) IB of galectin-9 following IP of human IgD and galectin-9 protein mix with control (ctrl) or anti-IgD antibodies. Prior to IP, the protein mix was supplemented with control PBS, glucose or lactose. (B) FCM of human IgD on human KU812 cells cultured for 30 min with control medium alone (ctrl), IgD or an IgD-galectin-9 complex formed by pre-incubating IgD with galectin-9 for 10 min. (C) FCM of human IgD or galectin-9 on KU812 cells cultured with IgD-galectin-9 in the presence of medium alone (ctrl), glucose or lactose for 30 min. (D) FCM of human IgD or CD44 on KU812 cells treated with scrambled (ctrl) or CD44-targeting small interfering RNA (siRNA) and later incubated with or without (ctrl) IgD-galectin-9. MFI, mean fluorescence intensity. (E) Confocal imaging of human basophils stained for IgD (blue), galectin-9 (green) and CD44 (red). Scale bar, 0.5 μm. (F) FCM of IgD on human basophils incubated with or without (ctrl) IgD-galectin-9 for 30 min. (G) ELISA of IL-4 from human basophils incubated with medium alone (ctrl) and with or without the IgD-galectin-9 complex in the presence or absence of IgD cross-linking by anti-IgD for 18 h. (H, I) FCM of IgD + basophils from the spleen or lung of WT C57BL/6, Lgals9 −/− (H) or Cd44 −/− (I) mice. (J, K) ELISA of serum IgG1 and IgE to NP from WT Balb/c (n=10), Lgals9 −/− (n=8) (J) or Cd44 −/− (n=10) (K) mice following s.c. immunization with NP-OVA and papain. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of anti-IgD treatment (day 0). (L) ELISA of serum total IgG1 and IgE from WT C57BL/6 controls (n=10) and Lgals9 −/− mice (n=8) following i.p. injection of anti-IgD. .

Techniques Used: Cell Culture, Small Interfering RNA, Incubation, Fluorescence, Imaging, Staining, Enzyme-linked Immunosorbent Assay, Mouse Assay, Concentration Assay, Injection

Ligation of Basophil-Bound IgD by Antigen Enhances IgG1 and IgE Responses (A) Schematics of i.p. immunization with NP-OVA and papain. (B) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of immunization (day 0). (C-D) FCM quantification of total and IL-4-expressing splenic PD-1 high CXCR5 high Tfh cells from WT Balb/c (n=5) or Ighd −/− (n=5) mice immunized as in (A). (E, F) ELISA of serum NP-specific IgG1 from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). BSA haptenated with 4 or 16 NPs was used to measure high-affinity (HA) and both HA and lowaffinity (LA) IgG1, respectively. (G) IFA of splenic tissue from immunized Balb/c mice stained for IgD (green), IgM (red) and nuclei (blue) following i.p. immunization as in (B). Inset: IgD + IgM − plasmablast next to IgD − IgM + plasmablast. FO, follicle. Original magnification, ×10 with ×2 enlargement. Scale bar, 50 μm. (H) ELISPOT of spleen ASCs expressing NP-specific IgD from WT Balb/c (n=5) or Ighd −/− (n=5) mice 3 d following i.p. immunization with PBS or NP-OVA and papain. (I) Schematics of i.v. reconstitution with NP-reactive IgD (NP-IgD) followed by i.p. immunization with NP-OVA and papain. (J) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c controls (n=10), Ighd −/− mice (n=10) or NP-IgD-reconstituted Ighd −/− mice (n=10) following i.p. immunization with NP-OVA and papain as in (I). (K) FCM of circulating FcεRI + IgE + basophils from WT Balb/c mice after i.v. injection of a control (ctrl) IgG2b antibody or a basophil-depleting anti-CD200R3 antibody. (L) ELISA of serum total IgD as well as serum NP-specific IgG1 and IgE in control (n=10) or basophil-depleted (n=10) WT Balb/c mice after i.p. immunization with NP-OVA and papain. .
Figure Legend Snippet: Ligation of Basophil-Bound IgD by Antigen Enhances IgG1 and IgE Responses (A) Schematics of i.p. immunization with NP-OVA and papain. (B) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of immunization (day 0). (C-D) FCM quantification of total and IL-4-expressing splenic PD-1 high CXCR5 high Tfh cells from WT Balb/c (n=5) or Ighd −/− (n=5) mice immunized as in (A). (E, F) ELISA of serum NP-specific IgG1 from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). BSA haptenated with 4 or 16 NPs was used to measure high-affinity (HA) and both HA and lowaffinity (LA) IgG1, respectively. (G) IFA of splenic tissue from immunized Balb/c mice stained for IgD (green), IgM (red) and nuclei (blue) following i.p. immunization as in (B). Inset: IgD + IgM − plasmablast next to IgD − IgM + plasmablast. FO, follicle. Original magnification, ×10 with ×2 enlargement. Scale bar, 50 μm. (H) ELISPOT of spleen ASCs expressing NP-specific IgD from WT Balb/c (n=5) or Ighd −/− (n=5) mice 3 d following i.p. immunization with PBS or NP-OVA and papain. (I) Schematics of i.v. reconstitution with NP-reactive IgD (NP-IgD) followed by i.p. immunization with NP-OVA and papain. (J) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c controls (n=10), Ighd −/− mice (n=10) or NP-IgD-reconstituted Ighd −/− mice (n=10) following i.p. immunization with NP-OVA and papain as in (I). (K) FCM of circulating FcεRI + IgE + basophils from WT Balb/c mice after i.v. injection of a control (ctrl) IgG2b antibody or a basophil-depleting anti-CD200R3 antibody. (L) ELISA of serum total IgD as well as serum NP-specific IgG1 and IgE in control (n=10) or basophil-depleted (n=10) WT Balb/c mice after i.p. immunization with NP-OVA and papain. .

Techniques Used: Ligation, Enzyme-linked Immunosorbent Assay, Mouse Assay, Concentration Assay, Expressing, Immunofluorescence, Staining, Enzyme-linked Immunospot, Injection

IgD Ligation by Antigen Induces Basophil Expression of IL-4 (A) Schematics of s.c. immunization with NP-OVA combined with control PBS, papain or NP-IgD. (B) Confocal imaging of CD169 (subcapsular sinus macrophage molecule, red), B220 (B cell molecule, blue) and Mcpt8 (YFP, yellow) from draining lymph node (DLN) of a C57BL/6 Mcpt8 YFP mouse immunized for 6 h as in (A). FO, follicle; dashed line, follicular border. Original magnification, ×5; rightbottom panel, ×40. Scale bars, 50 μm (top and bottom-left panels) or 5 μm (bottom-right panel). (C-E) FCM quantification of total YFP + or GFP + basophils and qRT-PCR quantification of Il4 transcripts encoding IL-4 from the DLN of C57BL/6 Mcpt8 YFP (n=5) mice (C, D) or Balb/c Il4 GFP (n=10) mice (E) 6 h following s.c. immunization as in (A). qRT-PCR results (D, bottom graph) are presented as relative expression (RE) compared to mRNA for glyceraldeheyde phosphate dehydrogenase (GAPDH). (F) Schematics of s.c. immunization with NP-OVA combined with papain, alum or CFA. (G) ELISA of serum NP-specific IgD from WT Balb/c mice (n=17) following s.c. immunization with NP-OVA and papain. PI, pre-immune (day −1 relatively to the onset of immunization (day 0). (H) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=12) or Ighd −/− (n=16) mice following s.c. immunization with NP-OVA and papain. (I) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=15) or Ighd −/− (n=15) mice following s.c. immunization with NP-OVA and alum. (J) ELISA of serum NP-specific IgG2a and IgG2b from Balb/c (n=10) or Ighd −/− (n=10) mice following s.c. immunization with NP-OVA and CFA. Data show one experiment of three with similar results (B, C) or summarize results from two experiments with 5–10 (D, E) or 10–17 (G-J) mice per experimental group. Results are presented as mean ± SEM; *p
Figure Legend Snippet: IgD Ligation by Antigen Induces Basophil Expression of IL-4 (A) Schematics of s.c. immunization with NP-OVA combined with control PBS, papain or NP-IgD. (B) Confocal imaging of CD169 (subcapsular sinus macrophage molecule, red), B220 (B cell molecule, blue) and Mcpt8 (YFP, yellow) from draining lymph node (DLN) of a C57BL/6 Mcpt8 YFP mouse immunized for 6 h as in (A). FO, follicle; dashed line, follicular border. Original magnification, ×5; rightbottom panel, ×40. Scale bars, 50 μm (top and bottom-left panels) or 5 μm (bottom-right panel). (C-E) FCM quantification of total YFP + or GFP + basophils and qRT-PCR quantification of Il4 transcripts encoding IL-4 from the DLN of C57BL/6 Mcpt8 YFP (n=5) mice (C, D) or Balb/c Il4 GFP (n=10) mice (E) 6 h following s.c. immunization as in (A). qRT-PCR results (D, bottom graph) are presented as relative expression (RE) compared to mRNA for glyceraldeheyde phosphate dehydrogenase (GAPDH). (F) Schematics of s.c. immunization with NP-OVA combined with papain, alum or CFA. (G) ELISA of serum NP-specific IgD from WT Balb/c mice (n=17) following s.c. immunization with NP-OVA and papain. PI, pre-immune (day −1 relatively to the onset of immunization (day 0). (H) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=12) or Ighd −/− (n=16) mice following s.c. immunization with NP-OVA and papain. (I) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=15) or Ighd −/− (n=15) mice following s.c. immunization with NP-OVA and alum. (J) ELISA of serum NP-specific IgG2a and IgG2b from Balb/c (n=10) or Ighd −/− (n=10) mice following s.c. immunization with NP-OVA and CFA. Data show one experiment of three with similar results (B, C) or summarize results from two experiments with 5–10 (D, E) or 10–17 (G-J) mice per experimental group. Results are presented as mean ± SEM; *p

Techniques Used: Ligation, Expressing, Imaging, Quantitative RT-PCR, Mouse Assay, Enzyme-linked Immunosorbent Assay

IgD Attenuates Acute Lung Inflammation Following Secondary Antigen Exposure (A) Schematics of i.p. OVA sensitization followed by a challenge consisting of four consecutive i.t. inoculations of OVA or control PBS. (B) ELISA of OVA-specific IgD and IgE from serum of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). Dashed line, maximum antibody concentration 4 hs after the last i.t. inoculation of PBS. (C) FCM quantitation of IgE on l basophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (D) Microscopic quantitation of total cells from the bronchoalveolar lavage (BAL) of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (E, F) FCM quantitation of CD49b + IgE + basophils from lungs of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). NTNB, non-T non-B. (G) ELISA of Mcpt8 from serum of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (H, I) FCM quantitation of CD45 + Siglec-F + eosinophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). Data summarize two experiments with at least five mice per experimental group. Results are presented as mean ± SEM; *p
Figure Legend Snippet: IgD Attenuates Acute Lung Inflammation Following Secondary Antigen Exposure (A) Schematics of i.p. OVA sensitization followed by a challenge consisting of four consecutive i.t. inoculations of OVA or control PBS. (B) ELISA of OVA-specific IgD and IgE from serum of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). Dashed line, maximum antibody concentration 4 hs after the last i.t. inoculation of PBS. (C) FCM quantitation of IgE on l basophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (D) Microscopic quantitation of total cells from the bronchoalveolar lavage (BAL) of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (E, F) FCM quantitation of CD49b + IgE + basophils from lungs of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). NTNB, non-T non-B. (G) ELISA of Mcpt8 from serum of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (H, I) FCM quantitation of CD45 + Siglec-F + eosinophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). Data summarize two experiments with at least five mice per experimental group. Results are presented as mean ± SEM; *p

Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay, Concentration Assay, Quantitation Assay

2) Product Images from "Blood biomarker for Parkinson disease: peptoids"

Article Title: Blood biomarker for Parkinson disease: peptoids

Journal: NPJ Parkinson's disease

doi: 10.1038/npjparkd.2016.12

On-bead magnetic screening. A one-bead one-compound (OBOC) library of thousands of unique peptoid compounds bound to TentaGel beads is incubated with control serum, here serum pooled from normal control subjects. The library is then incubated with anti-human IgG-labeled magnetic nanoparticles so that beads having bound IgG from the serum can be sorted out using a strong magnet. The library is initially depleted of beads that bind IgG from the control serum, and then incubated with target serum, here serum pooled from PD subjects. After incubation with the magnetic nanoparticles again, the newly magnetized beads, called ‘hits’, are isolated. Peptoid compounds are cleaved from each of the ‘hit’ beads and their sequences are assessed by MS/MS. These ‘hit’ compounds are then resynthesized and validated on ELISA plates for their ability to detect target IgG. (Reprinted by permission from Macmillan Publishers: from Zaman et al. 29 ). ELISA, enzyme-linked immunosorben assay.
Figure Legend Snippet: On-bead magnetic screening. A one-bead one-compound (OBOC) library of thousands of unique peptoid compounds bound to TentaGel beads is incubated with control serum, here serum pooled from normal control subjects. The library is then incubated with anti-human IgG-labeled magnetic nanoparticles so that beads having bound IgG from the serum can be sorted out using a strong magnet. The library is initially depleted of beads that bind IgG from the control serum, and then incubated with target serum, here serum pooled from PD subjects. After incubation with the magnetic nanoparticles again, the newly magnetized beads, called ‘hits’, are isolated. Peptoid compounds are cleaved from each of the ‘hit’ beads and their sequences are assessed by MS/MS. These ‘hit’ compounds are then resynthesized and validated on ELISA plates for their ability to detect target IgG. (Reprinted by permission from Macmillan Publishers: from Zaman et al. 29 ). ELISA, enzyme-linked immunosorben assay.

Techniques Used: Incubation, Labeling, Isolation, Mass Spectrometry, Enzyme-linked Immunosorbent Assay

3) Product Images from "Chimeric antigen receptor T cells secreting anti-PD-L1 antibodies more effectively regress renal cell carcinoma in a humanized mouse model"

Article Title: Chimeric antigen receptor T cells secreting anti-PD-L1 antibodies more effectively regress renal cell carcinoma in a humanized mouse model

Journal: Oncotarget

doi: 10.18632/oncotarget.9114

Chimeric antigen receptors (CAR) constructs for CD8+ T cells transduction ( A ) T cells were transduced with the lentiviruses to generate anti-CAIX CAR T cells, which are able to recognize CAIX positive RCC, and also secrete anti-PD-L1 IgG1 or IgG4 in the tumor microenvironment to block PD-1/PD-L1-induced T cell exhaustion. ( B ) Schematic representation of pHAGE lentiviral vectors encoding second-generation CARs fused with CD28 co-stimulatory endodomain. The anti-carbonic anhydrase IX (CAIX) or the Anti-B-cell maturation antigen (BCMA) scFv (as a negative control) were inserted after the eIFa promoter in order to express the CAR binding domain. The second cassette, after the Internal Ribosome Entry Site (IRES) sequence, encodes the secretable anti-PD-L1 IgG1 or IgG4 isotypes or the anti-severe acute respiratory syndrome (SARS) coronavirus IgG1 (negative control). LTR: long terminal repeat, eIFα: eukaryotic initiation factor alpha, scFv: single-chain variable fragment, C9 TAG: C9 peptide TETSQVAPA, IRES: Internal Ribosome Entry Site, WPRE: Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element. ( C ) Percentage of CAR T cells 14 days after transduction, representing the stable long-term expression of CAR by the integrated lentiviruses in CD8+ T cells. The CD8+ T cells were selected using Dynabeads CD8 Positive Isolation Kit (Life Technologies) and activated with Dynabeads Human T Cell Activator CD3/CD28 (Life Technologies) in the presence of IL-21 50 U/mL. IL-21 was added to the medium every 2 days. After 14 days, the CAR T cells were incubated with human CAIX-Fc or BCMA-Fc, followed by incubation with an APC conjugated anti-human Fc IgG (Southern Biotech) or goat-anti-mouse IgG Ab (Biolegend) and analyzed by flow cytometry. ( D ) Concentration of IgG secreted into the medium of transduced T cells evaluated by Human IgG ELISA Quantitation Set (Bethyl Laboratories). ( E ) Concentration of anti-PD-L1 antibodies in the supernatant of 293T Cells transduced with lentiviruses containing anti-CAIX or anti-BCMA CAR and anti-PD-L1 IgG1, anti-PD-L1 IgG4 or irrelevant anti-SARS IgG1 sequences. The antibodies in the supernatant were purified, biotinylated and incubated with 5 μg/mL of human PD-L1 pre-immobilized in the 96 wells MaxiSorp plate (Nunc). The biotinylated antibodies were detected by incubation with streptavidin-HRP and developed with TMB. The absorbance was read at λ = 450 nm. * P
Figure Legend Snippet: Chimeric antigen receptors (CAR) constructs for CD8+ T cells transduction ( A ) T cells were transduced with the lentiviruses to generate anti-CAIX CAR T cells, which are able to recognize CAIX positive RCC, and also secrete anti-PD-L1 IgG1 or IgG4 in the tumor microenvironment to block PD-1/PD-L1-induced T cell exhaustion. ( B ) Schematic representation of pHAGE lentiviral vectors encoding second-generation CARs fused with CD28 co-stimulatory endodomain. The anti-carbonic anhydrase IX (CAIX) or the Anti-B-cell maturation antigen (BCMA) scFv (as a negative control) were inserted after the eIFa promoter in order to express the CAR binding domain. The second cassette, after the Internal Ribosome Entry Site (IRES) sequence, encodes the secretable anti-PD-L1 IgG1 or IgG4 isotypes or the anti-severe acute respiratory syndrome (SARS) coronavirus IgG1 (negative control). LTR: long terminal repeat, eIFα: eukaryotic initiation factor alpha, scFv: single-chain variable fragment, C9 TAG: C9 peptide TETSQVAPA, IRES: Internal Ribosome Entry Site, WPRE: Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element. ( C ) Percentage of CAR T cells 14 days after transduction, representing the stable long-term expression of CAR by the integrated lentiviruses in CD8+ T cells. The CD8+ T cells were selected using Dynabeads CD8 Positive Isolation Kit (Life Technologies) and activated with Dynabeads Human T Cell Activator CD3/CD28 (Life Technologies) in the presence of IL-21 50 U/mL. IL-21 was added to the medium every 2 days. After 14 days, the CAR T cells were incubated with human CAIX-Fc or BCMA-Fc, followed by incubation with an APC conjugated anti-human Fc IgG (Southern Biotech) or goat-anti-mouse IgG Ab (Biolegend) and analyzed by flow cytometry. ( D ) Concentration of IgG secreted into the medium of transduced T cells evaluated by Human IgG ELISA Quantitation Set (Bethyl Laboratories). ( E ) Concentration of anti-PD-L1 antibodies in the supernatant of 293T Cells transduced with lentiviruses containing anti-CAIX or anti-BCMA CAR and anti-PD-L1 IgG1, anti-PD-L1 IgG4 or irrelevant anti-SARS IgG1 sequences. The antibodies in the supernatant were purified, biotinylated and incubated with 5 μg/mL of human PD-L1 pre-immobilized in the 96 wells MaxiSorp plate (Nunc). The biotinylated antibodies were detected by incubation with streptavidin-HRP and developed with TMB. The absorbance was read at λ = 450 nm. * P

Techniques Used: Construct, Transduction, Blocking Assay, Negative Control, Binding Assay, Sequencing, Expressing, Isolation, Incubation, Flow Cytometry, Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Purification

4) Product Images from "Interleukin-10-producing LAG3+ regulatory T cells are associated with disease activity and abatacept treatment in rheumatoid arthritis"

Article Title: Interleukin-10-producing LAG3+ regulatory T cells are associated with disease activity and abatacept treatment in rheumatoid arthritis

Journal: Arthritis Research & Therapy

doi: 10.1186/s13075-017-1309-x

Human CD4 + CD25 − LAG3 + T cells suppressed antibody production. a Gating strategy of fluorescence-activated cell sorting analysis for CD4 + CD25 − LAG3 + CD45RA − T cells (LAG3 + Tregs) and CD4 + CD25 + CD127 dim CD45RA − T cells (CD25 + Tregs). b IL-10, IFN-γ, and IL-17A protein levels in the culture supernatants after 3 days of incubation of the indicated T-cell subsets was determined by ELISA ( n = 6). c B cells and T FH cells were cocultured with the indicated subsets for 12 days in the presence of SEB stimulation, and total IgG, IgM, and IgA production was determined by ELISA ( n = 6). d B cells and T FH cells were cocultured with CD4 + CD25 − LAG3 + CD49b − T cells for 12 days in the presence of SEB stimulation, and total IgG was determined by ELISA ( n = 3). All error bars represent SD. * P
Figure Legend Snippet: Human CD4 + CD25 − LAG3 + T cells suppressed antibody production. a Gating strategy of fluorescence-activated cell sorting analysis for CD4 + CD25 − LAG3 + CD45RA − T cells (LAG3 + Tregs) and CD4 + CD25 + CD127 dim CD45RA − T cells (CD25 + Tregs). b IL-10, IFN-γ, and IL-17A protein levels in the culture supernatants after 3 days of incubation of the indicated T-cell subsets was determined by ELISA ( n = 6). c B cells and T FH cells were cocultured with the indicated subsets for 12 days in the presence of SEB stimulation, and total IgG, IgM, and IgA production was determined by ELISA ( n = 6). d B cells and T FH cells were cocultured with CD4 + CD25 − LAG3 + CD49b − T cells for 12 days in the presence of SEB stimulation, and total IgG was determined by ELISA ( n = 3). All error bars represent SD. * P

Techniques Used: Fluorescence, FACS, Incubation, Enzyme-linked Immunosorbent Assay

5) Product Images from "Protein structure shapes immunodominance in the CD4 T cell response to yellow fever vaccination"

Article Title: Protein structure shapes immunodominance in the CD4 T cell response to yellow fever vaccination

Journal: Scientific Reports

doi: 10.1038/s41598-017-09331-w

CXCR5 + and CXCR5 − CD4 T cell response to YF virus C, prM and E peptides. ( a ) CD4 + CD45Ra − memory T cells from PBMC of healthy donors were sorted into CXCR5 + (upper right) and CXCR5 − cells (lower left). ( b ) IgG concentration revealed by ELISA in supernatants of sorted autologous B cells cultured for 10 days with either CXCR5 + or CXCR5 − cells in 4 independent experiments. Statistical signifiance was determined with the two-way ANOVA. ( c ) Percentage of cytokine events in sorted CXCR5 + and CXCR5 − subsets contributed by C, prM and E peptides as determined by intracellular cytokine staining. ( d ) Ratios of results with E and C peptides obtained in individual donors. Statistical signifiance was determined with the Wilcoxon matched-pairs signed rank test.
Figure Legend Snippet: CXCR5 + and CXCR5 − CD4 T cell response to YF virus C, prM and E peptides. ( a ) CD4 + CD45Ra − memory T cells from PBMC of healthy donors were sorted into CXCR5 + (upper right) and CXCR5 − cells (lower left). ( b ) IgG concentration revealed by ELISA in supernatants of sorted autologous B cells cultured for 10 days with either CXCR5 + or CXCR5 − cells in 4 independent experiments. Statistical signifiance was determined with the two-way ANOVA. ( c ) Percentage of cytokine events in sorted CXCR5 + and CXCR5 − subsets contributed by C, prM and E peptides as determined by intracellular cytokine staining. ( d ) Ratios of results with E and C peptides obtained in individual donors. Statistical signifiance was determined with the Wilcoxon matched-pairs signed rank test.

Techniques Used: Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Staining

6) Product Images from "Utility of Humanized BLT Mice for Analysis of Dengue Virus Infection and Antiviral Drug Testing"

Article Title: Utility of Humanized BLT Mice for Analysis of Dengue Virus Infection and Antiviral Drug Testing

Journal: Journal of Virology

doi: 10.1128/JVI.03085-13

Human humoral immune responses in BLT-NOD/ SCID mice infected with DENV-2. (A) Total human IgM antibodies circulating in the bloodstream of DENV-2 Col-infected and control animals were measured by ELISA at days 7, 13, 25, and 32 postinfection. Black diamonds
Figure Legend Snippet: Human humoral immune responses in BLT-NOD/ SCID mice infected with DENV-2. (A) Total human IgM antibodies circulating in the bloodstream of DENV-2 Col-infected and control animals were measured by ELISA at days 7, 13, 25, and 32 postinfection. Black diamonds

Techniques Used: Mouse Assay, Infection, Enzyme-linked Immunosorbent Assay

7) Product Images from "Immunophenotypic characterization of CSF B cells in virus-associated neuroinflammatory diseases"

Article Title: Immunophenotypic characterization of CSF B cells in virus-associated neuroinflammatory diseases

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1007042

Antibody responses against HTLV-1 in CSF of HTLV-1-infected subjects. (A) Comparison of antibody responses against HTLV-1 Gag, Env and Tax in serum and CSF of ACs and HAM/TSP patients using Mann-Whitney Test. (B) Comparison of CSF/serum anti-HTLV-1 Gag, Env and Tax antibody ratio of HAM/TSP patients and ACs using Paired T test or Mann-Whitney Test. All the data were obtained from HAM/TSP patients (n = 44) and ACs (n = 4). The horizontal line represents the mean. (C) Correlation of ASCs in CSF B cells with anti-Gag, anti-Env and anti-Tax antibody index in HAM/TSP patients (n = 11) using Spearman’s rank correlation test.
Figure Legend Snippet: Antibody responses against HTLV-1 in CSF of HTLV-1-infected subjects. (A) Comparison of antibody responses against HTLV-1 Gag, Env and Tax in serum and CSF of ACs and HAM/TSP patients using Mann-Whitney Test. (B) Comparison of CSF/serum anti-HTLV-1 Gag, Env and Tax antibody ratio of HAM/TSP patients and ACs using Paired T test or Mann-Whitney Test. All the data were obtained from HAM/TSP patients (n = 44) and ACs (n = 4). The horizontal line represents the mean. (C) Correlation of ASCs in CSF B cells with anti-Gag, anti-Env and anti-Tax antibody index in HAM/TSP patients (n = 11) using Spearman’s rank correlation test.

Techniques Used: Infection, MANN-WHITNEY

Involvement of CD4 + CD25 + T cells with B cell help in CSF of HAM/TSP patients. (A) Comparison of HTLV-1 PVL in CSF of ACs (n = 7) and HAM/TSP patients (n = 36) using Mann-Whitney Test. (B) Correlation of HTLV-1 PVL with CD4 + CD25 + T cells and memory Tfh cells in HTLV-1-infected subjects using Spearman’s rank correlation test. (C) Comparison of IL-21 in CSF of HAM/TSP patients and ACs using unpaired t test. (D) Representative dot plots of IL-21 and Tax staining in CD4 + CD25 + T cells of a ND and a HAM/TSP patient after culture for 24 hours without any exogenous stimulation. (E) Detection of IL-21 in Tax-expressing CD4 + CD25 + T cells of HAM/TSP patients after culture for 24 hours without any exogenous stimulation. (F) Generation of ASCs subsets in B cells cultured with and without rhIL-21. Th e data were obtained from cultured B cells of NDs and HAM/TSP patients (n = 6). The horizontal line represents the mean. (G) Detection of human IgG in the B cell culture supernatants of NDs and HAM/TSP patients. (H) Detection of antibodies for HTLV-1 Gag and Tax in the B cell culture supernatants of HAM/TSP patients.
Figure Legend Snippet: Involvement of CD4 + CD25 + T cells with B cell help in CSF of HAM/TSP patients. (A) Comparison of HTLV-1 PVL in CSF of ACs (n = 7) and HAM/TSP patients (n = 36) using Mann-Whitney Test. (B) Correlation of HTLV-1 PVL with CD4 + CD25 + T cells and memory Tfh cells in HTLV-1-infected subjects using Spearman’s rank correlation test. (C) Comparison of IL-21 in CSF of HAM/TSP patients and ACs using unpaired t test. (D) Representative dot plots of IL-21 and Tax staining in CD4 + CD25 + T cells of a ND and a HAM/TSP patient after culture for 24 hours without any exogenous stimulation. (E) Detection of IL-21 in Tax-expressing CD4 + CD25 + T cells of HAM/TSP patients after culture for 24 hours without any exogenous stimulation. (F) Generation of ASCs subsets in B cells cultured with and without rhIL-21. Th e data were obtained from cultured B cells of NDs and HAM/TSP patients (n = 6). The horizontal line represents the mean. (G) Detection of human IgG in the B cell culture supernatants of NDs and HAM/TSP patients. (H) Detection of antibodies for HTLV-1 Gag and Tax in the B cell culture supernatants of HAM/TSP patients.

Techniques Used: MANN-WHITNEY, Infection, Staining, Expressing, Cell Culture

8) Product Images from "Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia"

Article Title: Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia

Journal: PLoS ONE

doi: 10.1371/journal.pone.0162973

Contents of total proteins, immunoglobulin G (IgG), IgA, IgM, albumin, and complement 3 (C3) and C4 fractions throughout the purification process.
Figure Legend Snippet: Contents of total proteins, immunoglobulin G (IgG), IgA, IgM, albumin, and complement 3 (C3) and C4 fractions throughout the purification process.

Techniques Used: Purification

9) Product Images from "Human Vγ9Vδ2-T Cells Synergize CD4+ T Follicular Helper Cells to Produce Influenza Virus-Specific Antibody"

Article Title: Human Vγ9Vδ2-T Cells Synergize CD4+ T Follicular Helper Cells to Produce Influenza Virus-Specific Antibody

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.00599

Vγ9Vδ2-T cells increased H9N2 virus-specific IgG and IgM productions in vivo . Specific immune cells-depleted and whole peripheral blood mononuclear cells (PBMC)-humanized mice were vaccinated with UV-inactivated H9N2 virus through i.p. pathway on day 0 and 7. Serum was collected on day 21 post the first vaccination. (A,B) Total IgG and IgM in serum (C,D) H9N2 virus-specific IgG and IgM in serum that treated with receptor destroyed enzyme (RDE) were determined by enzyme-linked immunosorbent assay. (E) H9N2-specific antibodies in serum that treated with RDE were determined by hemagglutination inhibition assay. The data shown were the represent of four independent experiments. (F) Representative histological analysis, immunohistological stainings for human CD4 T, B, and γδ-T cells in formalin-fixed paraffin-embedded spleen in humanized mice constructed with whole PBMCs after UV-H9N2 virus immunization. Scale bars = 100 μm. Bottom panels showed the higher-magnification views of the respective boxed areas in the top panels. Scale bars = 50 μm. The data shown are the mean ± SEM. * p
Figure Legend Snippet: Vγ9Vδ2-T cells increased H9N2 virus-specific IgG and IgM productions in vivo . Specific immune cells-depleted and whole peripheral blood mononuclear cells (PBMC)-humanized mice were vaccinated with UV-inactivated H9N2 virus through i.p. pathway on day 0 and 7. Serum was collected on day 21 post the first vaccination. (A,B) Total IgG and IgM in serum (C,D) H9N2 virus-specific IgG and IgM in serum that treated with receptor destroyed enzyme (RDE) were determined by enzyme-linked immunosorbent assay. (E) H9N2-specific antibodies in serum that treated with RDE were determined by hemagglutination inhibition assay. The data shown were the represent of four independent experiments. (F) Representative histological analysis, immunohistological stainings for human CD4 T, B, and γδ-T cells in formalin-fixed paraffin-embedded spleen in humanized mice constructed with whole PBMCs after UV-H9N2 virus immunization. Scale bars = 100 μm. Bottom panels showed the higher-magnification views of the respective boxed areas in the top panels. Scale bars = 50 μm. The data shown are the mean ± SEM. * p

Techniques Used: In Vivo, Mouse Assay, Enzyme-linked Immunosorbent Assay, HI Assay, Formalin-fixed Paraffin-Embedded, Construct

Vγ9Vδ2-T cells facilitated influenza virus-specific antibody production. Naïve CD4 T cells were cultured with UV-H9N2 virus-pulsed dendritic cells for 4–5 days, then, the activated CD4 T cells were cultured with CD19 + B cells from same donor with or without Vγ9Vδ2-T cells for another 7–10 days. (A,B) Total IgG and IgM in supernatant on day 7 were detected by enzyme-linked immunosorbent assay (ELISA). (C,D) H9N2 virus-specific IgG and IgM were detected by ELISA on day 7. Each dot means one donor. The data shown are the mean ± SEM. * p
Figure Legend Snippet: Vγ9Vδ2-T cells facilitated influenza virus-specific antibody production. Naïve CD4 T cells were cultured with UV-H9N2 virus-pulsed dendritic cells for 4–5 days, then, the activated CD4 T cells were cultured with CD19 + B cells from same donor with or without Vγ9Vδ2-T cells for another 7–10 days. (A,B) Total IgG and IgM in supernatant on day 7 were detected by enzyme-linked immunosorbent assay (ELISA). (C,D) H9N2 virus-specific IgG and IgM were detected by ELISA on day 7. Each dot means one donor. The data shown are the mean ± SEM. * p

Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

10) Product Images from "Mechanistic insights into the impairment of memory B cells and antibody production in the elderly"

Article Title: Mechanistic insights into the impairment of memory B cells and antibody production in the elderly

Journal: Age

doi: 10.1007/s11357-011-9371-9

Kinetics of B cell responses before and at 1 and 9 months after two-dose primary vaccination and at 1 and 9 months after booster. a , b Geometric means with 95% CI of plasma TBEV-specific IgG concentrations (ELISA) and c , d neutralizing
Figure Legend Snippet: Kinetics of B cell responses before and at 1 and 9 months after two-dose primary vaccination and at 1 and 9 months after booster. a , b Geometric means with 95% CI of plasma TBEV-specific IgG concentrations (ELISA) and c , d neutralizing

Techniques Used: Enzyme-linked Immunosorbent Assay

11) Product Images from "Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls"

Article Title: Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls

Journal: Viral Immunology

doi: 10.1089/vim.2018.0032

The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.
Figure Legend Snippet: The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.

Techniques Used: Concentration Assay

Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).
Figure Legend Snippet: Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).

Techniques Used:

12) Product Images from "A monoclonal antibody-based immunoassay to measure the antibody response against the repeat region of the circumsporozoite protein of Plasmodium falciparum"

Article Title: A monoclonal antibody-based immunoassay to measure the antibody response against the repeat region of the circumsporozoite protein of Plasmodium falciparum

Journal: Malaria Journal

doi: 10.1186/s12936-016-1596-8

Comparison of antibody concentrations in sera from different time points and treatment groups. In both studies sera were obtained after the second (PD2) and third (PD3) vaccine dose and antibody concentrations were measured with R32LR and MAL1C-competition ELISA. In study 1 results obtained in the subjects receiving three doses of RTS,S/AS01 (3xRTS,S/AS01) were compared with those generated in subjects vaccinated with one dose of Ad35.CS.01 followed by two doses of RTS,S/AS01. In study 2, subjects vaccinated with three standard doses of RTS,S/AS01 given with monthly intervals at month 0, 1 and 2 (012) were compared with subjects given a standard dose at months 0 and 1 and 1/5 th of the standard dose at month 5 (017). No significant differences were observed between the study groups or between 2 and 3 vaccine doses
Figure Legend Snippet: Comparison of antibody concentrations in sera from different time points and treatment groups. In both studies sera were obtained after the second (PD2) and third (PD3) vaccine dose and antibody concentrations were measured with R32LR and MAL1C-competition ELISA. In study 1 results obtained in the subjects receiving three doses of RTS,S/AS01 (3xRTS,S/AS01) were compared with those generated in subjects vaccinated with one dose of Ad35.CS.01 followed by two doses of RTS,S/AS01. In study 2, subjects vaccinated with three standard doses of RTS,S/AS01 given with monthly intervals at month 0, 1 and 2 (012) were compared with subjects given a standard dose at months 0 and 1 and 1/5 th of the standard dose at month 5 (017). No significant differences were observed between the study groups or between 2 and 3 vaccine doses

Techniques Used: Enzyme-linked Immunosorbent Assay, Generated

Correlation between R32LR ELISA and MAL1C-competition ELISA. Sera from participants in study 1 ( panel A ) and study 2 ( panel B ) taken after the second and third vaccine doses (immediately prior to mosquito bite challenge) were analysed with the standard R32LR ELISA and the MAL1C-competition ELISA. Results obtained with both assays are plotted and correlation is calculated with Pearson’s correlation
Figure Legend Snippet: Correlation between R32LR ELISA and MAL1C-competition ELISA. Sera from participants in study 1 ( panel A ) and study 2 ( panel B ) taken after the second and third vaccine doses (immediately prior to mosquito bite challenge) were analysed with the standard R32LR ELISA and the MAL1C-competition ELISA. Results obtained with both assays are plotted and correlation is calculated with Pearson’s correlation

Techniques Used: Enzyme-linked Immunosorbent Assay

Development of the MAL1C-competition ELISA. Optimal dilutions of B-MAL1C and streptavidin-HRP were defined using a checkerboard titration experiment ( a ). At a 1/16,000 dilution of streptavidin-HRP a sigmoidal curve was observed; this dilution was used for further experiments. B-MAL1C dilutions in the range between the maximal absorbance (plateau observed at 1/50,000) and half max (max/2 observed at 1/400,000) were explored in an inhibition set up using defined sera with high and low antibody content in R32LR ELISA assay ( b ) and the three monoclonal antibodies, MAL1C, MAL2A and MAL3B ( c ). The data shown in panels b and c were obtained with the highest serum concentrations (starting dilution 1/5) and with mAb concentrations of 5 µg/ml
Figure Legend Snippet: Development of the MAL1C-competition ELISA. Optimal dilutions of B-MAL1C and streptavidin-HRP were defined using a checkerboard titration experiment ( a ). At a 1/16,000 dilution of streptavidin-HRP a sigmoidal curve was observed; this dilution was used for further experiments. B-MAL1C dilutions in the range between the maximal absorbance (plateau observed at 1/50,000) and half max (max/2 observed at 1/400,000) were explored in an inhibition set up using defined sera with high and low antibody content in R32LR ELISA assay ( b ) and the three monoclonal antibodies, MAL1C, MAL2A and MAL3B ( c ). The data shown in panels b and c were obtained with the highest serum concentrations (starting dilution 1/5) and with mAb concentrations of 5 µg/ml

Techniques Used: Enzyme-linked Immunosorbent Assay, Titration, Inhibition

Correlation between protection from infection and antibody content, measured with R32LR and MAL1C-competition ELISA. Antibody concentrations in the sera obtained immediately before challenge with infected mosquito’s from participants at the study 1 (n = 46) and study 2 (n = 46) studies were measured with the R32LR ELISA and MAL1C-competition ELISA. Antibody levels in protected and non-protected vaccine recipients were compared and no significant differences were observed irrespective of the assay used
Figure Legend Snippet: Correlation between protection from infection and antibody content, measured with R32LR and MAL1C-competition ELISA. Antibody concentrations in the sera obtained immediately before challenge with infected mosquito’s from participants at the study 1 (n = 46) and study 2 (n = 46) studies were measured with the R32LR ELISA and MAL1C-competition ELISA. Antibody levels in protected and non-protected vaccine recipients were compared and no significant differences were observed irrespective of the assay used

Techniques Used: Infection, Enzyme-linked Immunosorbent Assay

13) Product Images from "Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls"

Article Title: Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls

Journal: Viral Immunology

doi: 10.1089/vim.2018.0032

The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.
Figure Legend Snippet: The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.

Techniques Used: Concentration Assay

Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).
Figure Legend Snippet: Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).

Techniques Used:

14) Product Images from "Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls"

Article Title: Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls

Journal: Viral Immunology

doi: 10.1089/vim.2018.0032

The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.
Figure Legend Snippet: The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.

Techniques Used: Concentration Assay

Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).
Figure Legend Snippet: Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).

Techniques Used:

15) Product Images from "Secreted IgD Amplifies Humoral T Helper-2 Responses by Binding Basophils Via Galectin-9 and CD44"

Article Title: Secreted IgD Amplifies Humoral T Helper-2 Responses by Binding Basophils Via Galectin-9 and CD44

Journal: Immunity

doi: 10.1016/j.immuni.2018.08.013

IgD Ligation by Antigen Elicits Basophil Activation (A) ELISA of serum IgD from WT Balb/c (n=29), WT C57BL/6 (n=15), Ighd −/− Balb/c (n=20) or Ighd −/− C57BL/6 (n=10) mice. (B) Left: FCM of CD49b and IgE on basophils (black gate) from circulating CD45 + NTNB cells of a WT Balb/c mouse. NTNB, non-T non-B. Right: FCM of IgD on circulating basophils (blue open profile) from a WT Balb/c mouse. Ctrl, isotype-matched control (gray solid profile). (C) FCM of IgD + basophils from peripheral blood, spleen, bone marrow (BM), lung or mesenteric lymph nodes (MLNs) of WT Balb/c mice (n=5). (D) Imaging FCM of IgD, FcεRI and CD49b from a representative viable Ghost Dye Violet 510 (GV510) − splenic basophil of a WT Balb/c mouse. Scale bar, 5 μm. (E) FCM quantification of FcεRI + CD49b + basophils from BM, blood, spleen or lung CD45 + NTNB cells of WT Balb/c (n=10) or Ighd −/− (n=10) mice. (F) FCM of Il4 -driven GFP expression in splenic or lung FcεRI + CD49b + basophils from WT Balb/c Il4 GFP (n=5) or Il4 GFP Ighd −/− (n=5) mice. MFI, mean fluorescence intensity. (G) ELISA of IL-4 and IL-13 from serum of WT Balb/c or Ighd −/− mice (n=10). (H) Schematics of i.v. reconstitution μMT or Rag2 −/− mice with NP-reactive IgD (NP-IgD) followed by i.p. injection of anti-IgD or i.n. inoculation of NP-OVA. (I) FCM of IgD on splenic FcεRI + CD49b + basophils from a μMT mouse before (ctrl) or after reconstitution as in (H). PBS, control phosphate buffer solution (PBS). (J, K) FCM quantification of total, CD200R3 + or IL-4 + basophils from the spleen of μMT C57BL/6 mice (n=10) (J) or the lungs of IgD-deficient Rag2 −/− (n=10) C57BL/6 mice (K) treated as in (H). (L) FCM quantification of total, IgD + , IgE + or IL-4 + basophils from the lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice 5 d following i.n. exposure to PBS or recombinant TSLP. (M) ELISA of serum IgD from Balb/c mice following i.v. injection of control empty or TSLP-encoding plasmids (n=10). .
Figure Legend Snippet: IgD Ligation by Antigen Elicits Basophil Activation (A) ELISA of serum IgD from WT Balb/c (n=29), WT C57BL/6 (n=15), Ighd −/− Balb/c (n=20) or Ighd −/− C57BL/6 (n=10) mice. (B) Left: FCM of CD49b and IgE on basophils (black gate) from circulating CD45 + NTNB cells of a WT Balb/c mouse. NTNB, non-T non-B. Right: FCM of IgD on circulating basophils (blue open profile) from a WT Balb/c mouse. Ctrl, isotype-matched control (gray solid profile). (C) FCM of IgD + basophils from peripheral blood, spleen, bone marrow (BM), lung or mesenteric lymph nodes (MLNs) of WT Balb/c mice (n=5). (D) Imaging FCM of IgD, FcεRI and CD49b from a representative viable Ghost Dye Violet 510 (GV510) − splenic basophil of a WT Balb/c mouse. Scale bar, 5 μm. (E) FCM quantification of FcεRI + CD49b + basophils from BM, blood, spleen or lung CD45 + NTNB cells of WT Balb/c (n=10) or Ighd −/− (n=10) mice. (F) FCM of Il4 -driven GFP expression in splenic or lung FcεRI + CD49b + basophils from WT Balb/c Il4 GFP (n=5) or Il4 GFP Ighd −/− (n=5) mice. MFI, mean fluorescence intensity. (G) ELISA of IL-4 and IL-13 from serum of WT Balb/c or Ighd −/− mice (n=10). (H) Schematics of i.v. reconstitution μMT or Rag2 −/− mice with NP-reactive IgD (NP-IgD) followed by i.p. injection of anti-IgD or i.n. inoculation of NP-OVA. (I) FCM of IgD on splenic FcεRI + CD49b + basophils from a μMT mouse before (ctrl) or after reconstitution as in (H). PBS, control phosphate buffer solution (PBS). (J, K) FCM quantification of total, CD200R3 + or IL-4 + basophils from the spleen of μMT C57BL/6 mice (n=10) (J) or the lungs of IgD-deficient Rag2 −/− (n=10) C57BL/6 mice (K) treated as in (H). (L) FCM quantification of total, IgD + , IgE + or IL-4 + basophils from the lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice 5 d following i.n. exposure to PBS or recombinant TSLP. (M) ELISA of serum IgD from Balb/c mice following i.v. injection of control empty or TSLP-encoding plasmids (n=10). .

Techniques Used: Ligation, Activation Assay, Enzyme-linked Immunosorbent Assay, Mouse Assay, Imaging, Expressing, Fluorescence, Injection, Recombinant

Basophil-Bound IgD Interacts with Galectin-9 and Its Ligation Induces Th2 Cell-Associated Cytokine Expression But Inhibits Cytoskeleton Remodeling (A) Schematics of i.p. anti-IgD treated mice i.v. reconstituted or not with secreted IgD. (B-C) ELISA of serum IgG1, IgE (B), IL-4 and IL-13 (C) from WT Balb/c (n=10) or Ighd −/− (n=10) mice in the presence or absence of i.v. reconstitution with secreted IgD (NP-IgD), followed by i.p. injection of anti-IgD. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to onset of anti-IgD treatment (day 0). (D) IFA of human tonsillar tissue stained for IgD (green), IgM (red), cytokeratin (purple) and nuclei (blue). EP, epithelium; FO, follicle; PC, plasma cell. Original magnification, ×20 (top) or ×63 (bottom). Scale bars, 50 μm (top) and 5 μm (bottom). (E) ELISA of IgD specific to α-s-casein, β-lactoglobulin (β-LGB) or α-lactalbumin (α-LAL) from plasma of FPIES children with (n=5) or without (n=5) dietary milk restrictions. (F) FCM of IgD (red open profile) bound to human tonsillar or circulating CD123 + FcεRI + basophils (red gate). Gray open profiles, control isotype-matched antibody with irrelevant binding activity. (G) Microarray analysis of genes expressed by human basophils upon IgD cross-linking. The Volcano plot represents genes differentially expressed by basophils treated with anti-IgD or a F(ab’)2 control antibody (ctrl). Red and blue dots, up-regulated and down-regulated genes, respectively; FC, fold change. (H) Heat map of coordinated gene sets identified by gene set enrichment analysis in human basophils treated as in (G). NES (normalized enrichment score) indicates correlation between individual gene sets. Positive correlation, NES > 0 (yellow gradient); negative correlation, NES
Figure Legend Snippet: Basophil-Bound IgD Interacts with Galectin-9 and Its Ligation Induces Th2 Cell-Associated Cytokine Expression But Inhibits Cytoskeleton Remodeling (A) Schematics of i.p. anti-IgD treated mice i.v. reconstituted or not with secreted IgD. (B-C) ELISA of serum IgG1, IgE (B), IL-4 and IL-13 (C) from WT Balb/c (n=10) or Ighd −/− (n=10) mice in the presence or absence of i.v. reconstitution with secreted IgD (NP-IgD), followed by i.p. injection of anti-IgD. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to onset of anti-IgD treatment (day 0). (D) IFA of human tonsillar tissue stained for IgD (green), IgM (red), cytokeratin (purple) and nuclei (blue). EP, epithelium; FO, follicle; PC, plasma cell. Original magnification, ×20 (top) or ×63 (bottom). Scale bars, 50 μm (top) and 5 μm (bottom). (E) ELISA of IgD specific to α-s-casein, β-lactoglobulin (β-LGB) or α-lactalbumin (α-LAL) from plasma of FPIES children with (n=5) or without (n=5) dietary milk restrictions. (F) FCM of IgD (red open profile) bound to human tonsillar or circulating CD123 + FcεRI + basophils (red gate). Gray open profiles, control isotype-matched antibody with irrelevant binding activity. (G) Microarray analysis of genes expressed by human basophils upon IgD cross-linking. The Volcano plot represents genes differentially expressed by basophils treated with anti-IgD or a F(ab’)2 control antibody (ctrl). Red and blue dots, up-regulated and down-regulated genes, respectively; FC, fold change. (H) Heat map of coordinated gene sets identified by gene set enrichment analysis in human basophils treated as in (G). NES (normalized enrichment score) indicates correlation between individual gene sets. Positive correlation, NES > 0 (yellow gradient); negative correlation, NES

Techniques Used: Ligation, Expressing, Mouse Assay, Enzyme-linked Immunosorbent Assay, Injection, Concentration Assay, Immunofluorescence, Staining, Binding Assay, Activity Assay, Microarray

IgD Binds to Basophils through Galectin-9 and CD44 (A) IB of galectin-9 following IP of human IgD and galectin-9 protein mix with control (ctrl) or anti-IgD antibodies. Prior to IP, the protein mix was supplemented with control PBS, glucose or lactose. (B) FCM of human IgD on human KU812 cells cultured for 30 min with control medium alone (ctrl), IgD or an IgD-galectin-9 complex formed by pre-incubating IgD with galectin-9 for 10 min. (C) FCM of human IgD or galectin-9 on KU812 cells cultured with IgD-galectin-9 in the presence of medium alone (ctrl), glucose or lactose for 30 min. (D) FCM of human IgD or CD44 on KU812 cells treated with scrambled (ctrl) or CD44-targeting small interfering RNA (siRNA) and later incubated with or without (ctrl) IgD-galectin-9. MFI, mean fluorescence intensity. (E) Confocal imaging of human basophils stained for IgD (blue), galectin-9 (green) and CD44 (red). Scale bar, 0.5 μm. (F) FCM of IgD on human basophils incubated with or without (ctrl) IgD-galectin-9 for 30 min. (G) ELISA of IL-4 from human basophils incubated with medium alone (ctrl) and with or without the IgD-galectin-9 complex in the presence or absence of IgD cross-linking by anti-IgD for 18 h. (H, I) FCM of IgD + basophils from the spleen or lung of WT C57BL/6, Lgals9 −/− (H) or Cd44 −/− (I) mice. (J, K) ELISA of serum IgG1 and IgE to NP from WT Balb/c (n=10), Lgals9 −/− (n=8) (J) or Cd44 −/− (n=10) (K) mice following s.c. immunization with NP-OVA and papain. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of anti-IgD treatment (day 0). (L) ELISA of serum total IgG1 and IgE from WT C57BL/6 controls (n=10) and Lgals9 −/− mice (n=8) following i.p. injection of anti-IgD. .
Figure Legend Snippet: IgD Binds to Basophils through Galectin-9 and CD44 (A) IB of galectin-9 following IP of human IgD and galectin-9 protein mix with control (ctrl) or anti-IgD antibodies. Prior to IP, the protein mix was supplemented with control PBS, glucose or lactose. (B) FCM of human IgD on human KU812 cells cultured for 30 min with control medium alone (ctrl), IgD or an IgD-galectin-9 complex formed by pre-incubating IgD with galectin-9 for 10 min. (C) FCM of human IgD or galectin-9 on KU812 cells cultured with IgD-galectin-9 in the presence of medium alone (ctrl), glucose or lactose for 30 min. (D) FCM of human IgD or CD44 on KU812 cells treated with scrambled (ctrl) or CD44-targeting small interfering RNA (siRNA) and later incubated with or without (ctrl) IgD-galectin-9. MFI, mean fluorescence intensity. (E) Confocal imaging of human basophils stained for IgD (blue), galectin-9 (green) and CD44 (red). Scale bar, 0.5 μm. (F) FCM of IgD on human basophils incubated with or without (ctrl) IgD-galectin-9 for 30 min. (G) ELISA of IL-4 from human basophils incubated with medium alone (ctrl) and with or without the IgD-galectin-9 complex in the presence or absence of IgD cross-linking by anti-IgD for 18 h. (H, I) FCM of IgD + basophils from the spleen or lung of WT C57BL/6, Lgals9 −/− (H) or Cd44 −/− (I) mice. (J, K) ELISA of serum IgG1 and IgE to NP from WT Balb/c (n=10), Lgals9 −/− (n=8) (J) or Cd44 −/− (n=10) (K) mice following s.c. immunization with NP-OVA and papain. Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of anti-IgD treatment (day 0). (L) ELISA of serum total IgG1 and IgE from WT C57BL/6 controls (n=10) and Lgals9 −/− mice (n=8) following i.p. injection of anti-IgD. .

Techniques Used: Cell Culture, Small Interfering RNA, Incubation, Fluorescence, Imaging, Staining, Enzyme-linked Immunosorbent Assay, Mouse Assay, Concentration Assay, Injection

Ligation of Basophil-Bound IgD by Antigen Enhances IgG1 and IgE Responses (A) Schematics of i.p. immunization with NP-OVA and papain. (B) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of immunization (day 0). (C-D) FCM quantification of total and IL-4-expressing splenic PD-1 high CXCR5 high Tfh cells from WT Balb/c (n=5) or Ighd −/− (n=5) mice immunized as in (A). (E, F) ELISA of serum NP-specific IgG1 from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). BSA haptenated with 4 or 16 NPs was used to measure high-affinity (HA) and both HA and lowaffinity (LA) IgG1, respectively. (G) IFA of splenic tissue from immunized Balb/c mice stained for IgD (green), IgM (red) and nuclei (blue) following i.p. immunization as in (B). Inset: IgD + IgM − plasmablast next to IgD − IgM + plasmablast. FO, follicle. Original magnification, ×10 with ×2 enlargement. Scale bar, 50 μm. (H) ELISPOT of spleen ASCs expressing NP-specific IgD from WT Balb/c (n=5) or Ighd −/− (n=5) mice 3 d following i.p. immunization with PBS or NP-OVA and papain. (I) Schematics of i.v. reconstitution with NP-reactive IgD (NP-IgD) followed by i.p. immunization with NP-OVA and papain. (J) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c controls (n=10), Ighd −/− mice (n=10) or NP-IgD-reconstituted Ighd −/− mice (n=10) following i.p. immunization with NP-OVA and papain as in (I). (K) FCM of circulating FcεRI + IgE + basophils from WT Balb/c mice after i.v. injection of a control (ctrl) IgG2b antibody or a basophil-depleting anti-CD200R3 antibody. (L) ELISA of serum total IgD as well as serum NP-specific IgG1 and IgE in control (n=10) or basophil-depleted (n=10) WT Balb/c mice after i.p. immunization with NP-OVA and papain. .
Figure Legend Snippet: Ligation of Basophil-Bound IgD by Antigen Enhances IgG1 and IgE Responses (A) Schematics of i.p. immunization with NP-OVA and papain. (B) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). Dashed line, maximum antibody concentration in pre-immune (PI) mice. PI, day −1 relatively to the onset of immunization (day 0). (C-D) FCM quantification of total and IL-4-expressing splenic PD-1 high CXCR5 high Tfh cells from WT Balb/c (n=5) or Ighd −/− (n=5) mice immunized as in (A). (E, F) ELISA of serum NP-specific IgG1 from WT Balb/c (n=10) or Ighd −/− (n=10) mice immunized as in (A). BSA haptenated with 4 or 16 NPs was used to measure high-affinity (HA) and both HA and lowaffinity (LA) IgG1, respectively. (G) IFA of splenic tissue from immunized Balb/c mice stained for IgD (green), IgM (red) and nuclei (blue) following i.p. immunization as in (B). Inset: IgD + IgM − plasmablast next to IgD − IgM + plasmablast. FO, follicle. Original magnification, ×10 with ×2 enlargement. Scale bar, 50 μm. (H) ELISPOT of spleen ASCs expressing NP-specific IgD from WT Balb/c (n=5) or Ighd −/− (n=5) mice 3 d following i.p. immunization with PBS or NP-OVA and papain. (I) Schematics of i.v. reconstitution with NP-reactive IgD (NP-IgD) followed by i.p. immunization with NP-OVA and papain. (J) ELISA of serum OVA-specific IgG1 and IgE from WT Balb/c controls (n=10), Ighd −/− mice (n=10) or NP-IgD-reconstituted Ighd −/− mice (n=10) following i.p. immunization with NP-OVA and papain as in (I). (K) FCM of circulating FcεRI + IgE + basophils from WT Balb/c mice after i.v. injection of a control (ctrl) IgG2b antibody or a basophil-depleting anti-CD200R3 antibody. (L) ELISA of serum total IgD as well as serum NP-specific IgG1 and IgE in control (n=10) or basophil-depleted (n=10) WT Balb/c mice after i.p. immunization with NP-OVA and papain. .

Techniques Used: Ligation, Enzyme-linked Immunosorbent Assay, Mouse Assay, Concentration Assay, Expressing, Immunofluorescence, Staining, Enzyme-linked Immunospot, Injection

IgD Ligation by Antigen Induces Basophil Expression of IL-4 (A) Schematics of s.c. immunization with NP-OVA combined with control PBS, papain or NP-IgD. (B) Confocal imaging of CD169 (subcapsular sinus macrophage molecule, red), B220 (B cell molecule, blue) and Mcpt8 (YFP, yellow) from draining lymph node (DLN) of a C57BL/6 Mcpt8 YFP mouse immunized for 6 h as in (A). FO, follicle; dashed line, follicular border. Original magnification, ×5; rightbottom panel, ×40. Scale bars, 50 μm (top and bottom-left panels) or 5 μm (bottom-right panel). (C-E) FCM quantification of total YFP + or GFP + basophils and qRT-PCR quantification of Il4 transcripts encoding IL-4 from the DLN of C57BL/6 Mcpt8 YFP (n=5) mice (C, D) or Balb/c Il4 GFP (n=10) mice (E) 6 h following s.c. immunization as in (A). qRT-PCR results (D, bottom graph) are presented as relative expression (RE) compared to mRNA for glyceraldeheyde phosphate dehydrogenase (GAPDH). (F) Schematics of s.c. immunization with NP-OVA combined with papain, alum or CFA. (G) ELISA of serum NP-specific IgD from WT Balb/c mice (n=17) following s.c. immunization with NP-OVA and papain. PI, pre-immune (day −1 relatively to the onset of immunization (day 0). (H) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=12) or Ighd −/− (n=16) mice following s.c. immunization with NP-OVA and papain. (I) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=15) or Ighd −/− (n=15) mice following s.c. immunization with NP-OVA and alum. (J) ELISA of serum NP-specific IgG2a and IgG2b from Balb/c (n=10) or Ighd −/− (n=10) mice following s.c. immunization with NP-OVA and CFA. Data show one experiment of three with similar results (B, C) or summarize results from two experiments with 5–10 (D, E) or 10–17 (G-J) mice per experimental group. Results are presented as mean ± SEM; *p
Figure Legend Snippet: IgD Ligation by Antigen Induces Basophil Expression of IL-4 (A) Schematics of s.c. immunization with NP-OVA combined with control PBS, papain or NP-IgD. (B) Confocal imaging of CD169 (subcapsular sinus macrophage molecule, red), B220 (B cell molecule, blue) and Mcpt8 (YFP, yellow) from draining lymph node (DLN) of a C57BL/6 Mcpt8 YFP mouse immunized for 6 h as in (A). FO, follicle; dashed line, follicular border. Original magnification, ×5; rightbottom panel, ×40. Scale bars, 50 μm (top and bottom-left panels) or 5 μm (bottom-right panel). (C-E) FCM quantification of total YFP + or GFP + basophils and qRT-PCR quantification of Il4 transcripts encoding IL-4 from the DLN of C57BL/6 Mcpt8 YFP (n=5) mice (C, D) or Balb/c Il4 GFP (n=10) mice (E) 6 h following s.c. immunization as in (A). qRT-PCR results (D, bottom graph) are presented as relative expression (RE) compared to mRNA for glyceraldeheyde phosphate dehydrogenase (GAPDH). (F) Schematics of s.c. immunization with NP-OVA combined with papain, alum or CFA. (G) ELISA of serum NP-specific IgD from WT Balb/c mice (n=17) following s.c. immunization with NP-OVA and papain. PI, pre-immune (day −1 relatively to the onset of immunization (day 0). (H) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=12) or Ighd −/− (n=16) mice following s.c. immunization with NP-OVA and papain. (I) ELISA of serum NP-specific IgG1 and IgE from WT Balb/c (n=15) or Ighd −/− (n=15) mice following s.c. immunization with NP-OVA and alum. (J) ELISA of serum NP-specific IgG2a and IgG2b from Balb/c (n=10) or Ighd −/− (n=10) mice following s.c. immunization with NP-OVA and CFA. Data show one experiment of three with similar results (B, C) or summarize results from two experiments with 5–10 (D, E) or 10–17 (G-J) mice per experimental group. Results are presented as mean ± SEM; *p

Techniques Used: Ligation, Expressing, Imaging, Quantitative RT-PCR, Mouse Assay, Enzyme-linked Immunosorbent Assay

IgD Attenuates Acute Lung Inflammation Following Secondary Antigen Exposure (A) Schematics of i.p. OVA sensitization followed by a challenge consisting of four consecutive i.t. inoculations of OVA or control PBS. (B) ELISA of OVA-specific IgD and IgE from serum of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). Dashed line, maximum antibody concentration 4 hs after the last i.t. inoculation of PBS. (C) FCM quantitation of IgE on l basophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (D) Microscopic quantitation of total cells from the bronchoalveolar lavage (BAL) of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (E, F) FCM quantitation of CD49b + IgE + basophils from lungs of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). NTNB, non-T non-B. (G) ELISA of Mcpt8 from serum of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (H, I) FCM quantitation of CD45 + Siglec-F + eosinophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). Data summarize two experiments with at least five mice per experimental group. Results are presented as mean ± SEM; *p
Figure Legend Snippet: IgD Attenuates Acute Lung Inflammation Following Secondary Antigen Exposure (A) Schematics of i.p. OVA sensitization followed by a challenge consisting of four consecutive i.t. inoculations of OVA or control PBS. (B) ELISA of OVA-specific IgD and IgE from serum of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). Dashed line, maximum antibody concentration 4 hs after the last i.t. inoculation of PBS. (C) FCM quantitation of IgE on l basophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (D) Microscopic quantitation of total cells from the bronchoalveolar lavage (BAL) of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (E, F) FCM quantitation of CD49b + IgE + basophils from lungs of WT Balb/c (n=10) or Ighd −/− (n=10) mice treated as in (A). NTNB, non-T non-B. (G) ELISA of Mcpt8 from serum of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). (H, I) FCM quantitation of CD45 + Siglec-F + eosinophils from lungs of WT Balb/c (n=5) or Ighd −/− (n=5) mice treated as in (A). Data summarize two experiments with at least five mice per experimental group. Results are presented as mean ± SEM; *p

Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay, Concentration Assay, Quantitation Assay

16) Product Images from "Thyrotropin Receptor Antibody (TRAb)-IgM Levels Are Markedly Higher Than TRAb-IgG Levels in Graves' Disease Patients and Controls, and TRAb-IgM Production Is Related to Epstein–Barr Virus Reactivation"

Article Title: Thyrotropin Receptor Antibody (TRAb)-IgM Levels Are Markedly Higher Than TRAb-IgG Levels in Graves' Disease Patients and Controls, and TRAb-IgM Production Is Related to Epstein–Barr Virus Reactivation

Journal: Viral Immunology

doi: 10.1089/vim.2016.0043

Difference in serum TRAb levels between Graves' disease patients and controls. (a) Serum TRAb-IgG levels are significantly higher in Graves' disease patients than in controls (healthy subjects). (b) Serum TRAb-IgM levels are also significantly higher in Graves' disease patients than in controls. (a) p
Figure Legend Snippet: Difference in serum TRAb levels between Graves' disease patients and controls. (a) Serum TRAb-IgG levels are significantly higher in Graves' disease patients than in controls (healthy subjects). (b) Serum TRAb-IgM levels are also significantly higher in Graves' disease patients than in controls. (a) p

Techniques Used:

Immunoglobulin isotype difference in serum total Ig and TRAb levels in Graves' disease patients. The ratio of IgG to IgM between total Ig and TRAb was contrastive. * p
Figure Legend Snippet: Immunoglobulin isotype difference in serum total Ig and TRAb levels in Graves' disease patients. The ratio of IgG to IgM between total Ig and TRAb was contrastive. * p

Techniques Used:

Difference in serum TRAb-IgM levels between the high EBV Ab group and others. Serum TRAb-IgM levels are significantly higher in the high EBV Ab group (EBV reactivation) than in others ( p = 0.030*, the two-sample t -test). High EBV Ab represents samples with a high antibody index in EA-IgG and VCA-IgG, suggesting EBV reactivation. Others are constituted samples, except for cases with high EBV Ab levels. Ab, antibody; EA, early antigen; EBV, Epstein–Barr virus; VCA, virus capsid antigen.
Figure Legend Snippet: Difference in serum TRAb-IgM levels between the high EBV Ab group and others. Serum TRAb-IgM levels are significantly higher in the high EBV Ab group (EBV reactivation) than in others ( p = 0.030*, the two-sample t -test). High EBV Ab represents samples with a high antibody index in EA-IgG and VCA-IgG, suggesting EBV reactivation. Others are constituted samples, except for cases with high EBV Ab levels. Ab, antibody; EA, early antigen; EBV, Epstein–Barr virus; VCA, virus capsid antigen.

Techniques Used:

17) Product Images from "Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls"

Article Title: Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls

Journal: Viral Immunology

doi: 10.1089/vim.2018.0032

The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.
Figure Legend Snippet: The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.

Techniques Used: Concentration Assay

Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).
Figure Legend Snippet: Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).

Techniques Used:

18) Product Images from "A Highly Conserved Region Between Amino Acids 221 and 266 of Dengue Virus Non-Structural Protein 1 Is a Major Epitope Region in Infected Patients"

Article Title: A Highly Conserved Region Between Amino Acids 221 and 266 of Dengue Virus Non-Structural Protein 1 Is a Major Epitope Region in Infected Patients

Journal: The American Journal of Tropical Medicine and Hygiene

doi: 10.4269/ajtmh.13-0624

Isotyping and IgG subclass examination. ( A ) Hybridoma culture supernatants were tested by ELISA to determine the isotypes of the HuMAbs. Each ELISA included three positive controls (Ctrl; Ctrl IgA, Ctrl IgG, and Ctrl IgM). ( B ) IgG subclasses were determined
Figure Legend Snippet: Isotyping and IgG subclass examination. ( A ) Hybridoma culture supernatants were tested by ELISA to determine the isotypes of the HuMAbs. Each ELISA included three positive controls (Ctrl; Ctrl IgA, Ctrl IgG, and Ctrl IgM). ( B ) IgG subclasses were determined

Techniques Used: Enzyme-linked Immunosorbent Assay

19) Product Images from "Immunophenotypic characterization of CSF B cells in virus-associated neuroinflammatory diseases"

Article Title: Immunophenotypic characterization of CSF B cells in virus-associated neuroinflammatory diseases

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1007042

Antibody responses against HTLV-1 in CSF of HTLV-1-infected subjects. (A) Comparison of antibody responses against HTLV-1 Gag, Env and Tax in serum and CSF of ACs and HAM/TSP patients using Mann-Whitney Test. (B) Comparison of CSF/serum anti-HTLV-1 Gag, Env and Tax antibody ratio of HAM/TSP patients and ACs using Paired T test or Mann-Whitney Test. All the data were obtained from HAM/TSP patients (n = 44) and ACs (n = 4). The horizontal line represents the mean. (C) Correlation of ASCs in CSF B cells with anti-Gag, anti-Env and anti-Tax antibody index in HAM/TSP patients (n = 11) using Spearman’s rank correlation test.
Figure Legend Snippet: Antibody responses against HTLV-1 in CSF of HTLV-1-infected subjects. (A) Comparison of antibody responses against HTLV-1 Gag, Env and Tax in serum and CSF of ACs and HAM/TSP patients using Mann-Whitney Test. (B) Comparison of CSF/serum anti-HTLV-1 Gag, Env and Tax antibody ratio of HAM/TSP patients and ACs using Paired T test or Mann-Whitney Test. All the data were obtained from HAM/TSP patients (n = 44) and ACs (n = 4). The horizontal line represents the mean. (C) Correlation of ASCs in CSF B cells with anti-Gag, anti-Env and anti-Tax antibody index in HAM/TSP patients (n = 11) using Spearman’s rank correlation test.

Techniques Used: Infection, MANN-WHITNEY

Involvement of CD4 + CD25 + T cells with B cell help in CSF of HAM/TSP patients. (A) Comparison of HTLV-1 PVL in CSF of ACs (n = 7) and HAM/TSP patients (n = 36) using Mann-Whitney Test. (B) Correlation of HTLV-1 PVL with CD4 + CD25 + T cells and memory Tfh cells in HTLV-1-infected subjects using Spearman’s rank correlation test. (C) Comparison of IL-21 in CSF of HAM/TSP patients and ACs using unpaired t test. (D) Representative dot plots of IL-21 and Tax staining in CD4 + CD25 + T cells of a ND and a HAM/TSP patient after culture for 24 hours without any exogenous stimulation. (E) Detection of IL-21 in Tax-expressing CD4 + CD25 + T cells of HAM/TSP patients after culture for 24 hours without any exogenous stimulation. (F) Generation of ASCs subsets in B cells cultured with and without rhIL-21. The data were obtained from cultured B cells of NDs and HAM/TSP patients (n = 6). The horizontal line represents the mean. (G) Detection of human IgG in the B cell culture supernatants of NDs and HAM/TSP patients. (H) Detection of antibodies for HTLV-1 Gag and Tax in the B cell culture supernatants of HAM/TSP patients.
Figure Legend Snippet: Involvement of CD4 + CD25 + T cells with B cell help in CSF of HAM/TSP patients. (A) Comparison of HTLV-1 PVL in CSF of ACs (n = 7) and HAM/TSP patients (n = 36) using Mann-Whitney Test. (B) Correlation of HTLV-1 PVL with CD4 + CD25 + T cells and memory Tfh cells in HTLV-1-infected subjects using Spearman’s rank correlation test. (C) Comparison of IL-21 in CSF of HAM/TSP patients and ACs using unpaired t test. (D) Representative dot plots of IL-21 and Tax staining in CD4 + CD25 + T cells of a ND and a HAM/TSP patient after culture for 24 hours without any exogenous stimulation. (E) Detection of IL-21 in Tax-expressing CD4 + CD25 + T cells of HAM/TSP patients after culture for 24 hours without any exogenous stimulation. (F) Generation of ASCs subsets in B cells cultured with and without rhIL-21. The data were obtained from cultured B cells of NDs and HAM/TSP patients (n = 6). The horizontal line represents the mean. (G) Detection of human IgG in the B cell culture supernatants of NDs and HAM/TSP patients. (H) Detection of antibodies for HTLV-1 Gag and Tax in the B cell culture supernatants of HAM/TSP patients.

Techniques Used: MANN-WHITNEY, Infection, Staining, Expressing, Cell Culture

20) Product Images from "Human Vγ9Vδ2-T Cells Synergize CD4+ T Follicular Helper Cells to Produce Influenza Virus-Specific Antibody"

Article Title: Human Vγ9Vδ2-T Cells Synergize CD4+ T Follicular Helper Cells to Produce Influenza Virus-Specific Antibody

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.00599

Vγ9Vδ2-T cells increased H9N2 virus-specific IgG and IgM productions in vivo . Specific immune cells-depleted and whole peripheral blood mononuclear cells (PBMC)-humanized mice were vaccinated with UV-inactivated H9N2 virus through i.p. pathway on day 0 and 7. Serum was collected on day 21 post the first vaccination. (A,B) Total IgG and IgM in serum (C,D) H9N2 virus-specific IgG and IgM in serum that treated with receptor destroyed enzyme (RDE) were determined by enzyme-linked immunosorbent assay. (E) H9N2-specific antibodies in serum that treated with RDE were determined by hemagglutination inhibition assay. The data shown were the represent of four independent experiments. (F) Representative histological analysis, immunohistological stainings for human CD4 T, B, and γδ-T cells in formalin-fixed paraffin-embedded spleen in humanized mice constructed with whole PBMCs after UV-H9N2 virus immunization. Scale bars = 100 μm. Bottom panels showed the higher-magnification views of the respective boxed areas in the top panels. Scale bars = 50 μm. The data shown are the mean ± SEM. * p
Figure Legend Snippet: Vγ9Vδ2-T cells increased H9N2 virus-specific IgG and IgM productions in vivo . Specific immune cells-depleted and whole peripheral blood mononuclear cells (PBMC)-humanized mice were vaccinated with UV-inactivated H9N2 virus through i.p. pathway on day 0 and 7. Serum was collected on day 21 post the first vaccination. (A,B) Total IgG and IgM in serum (C,D) H9N2 virus-specific IgG and IgM in serum that treated with receptor destroyed enzyme (RDE) were determined by enzyme-linked immunosorbent assay. (E) H9N2-specific antibodies in serum that treated with RDE were determined by hemagglutination inhibition assay. The data shown were the represent of four independent experiments. (F) Representative histological analysis, immunohistological stainings for human CD4 T, B, and γδ-T cells in formalin-fixed paraffin-embedded spleen in humanized mice constructed with whole PBMCs after UV-H9N2 virus immunization. Scale bars = 100 μm. Bottom panels showed the higher-magnification views of the respective boxed areas in the top panels. Scale bars = 50 μm. The data shown are the mean ± SEM. * p

Techniques Used: In Vivo, Mouse Assay, Enzyme-linked Immunosorbent Assay, HI Assay, Formalin-fixed Paraffin-Embedded, Construct

Vγ9Vδ2-T cells facilitated influenza virus-specific antibody production. Naïve CD4 T cells were cultured with UV-H9N2 virus-pulsed dendritic cells for 4–5 days, then, the activated CD4 T cells were cultured with CD19 + B cells from same donor with or without Vγ9Vδ2-T cells for another 7–10 days. (A,B) Total IgG and IgM in supernatant on day 7 were detected by enzyme-linked immunosorbent assay (ELISA). (C,D) H9N2 virus-specific IgG and IgM were detected by ELISA on day 7. Each dot means one donor. The data shown are the mean ± SEM. * p
Figure Legend Snippet: Vγ9Vδ2-T cells facilitated influenza virus-specific antibody production. Naïve CD4 T cells were cultured with UV-H9N2 virus-pulsed dendritic cells for 4–5 days, then, the activated CD4 T cells were cultured with CD19 + B cells from same donor with or without Vγ9Vδ2-T cells for another 7–10 days. (A,B) Total IgG and IgM in supernatant on day 7 were detected by enzyme-linked immunosorbent assay (ELISA). (C,D) H9N2 virus-specific IgG and IgM were detected by ELISA on day 7. Each dot means one donor. The data shown are the mean ± SEM. * p

Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

21) Product Images from "Immunophenotypic characterization of CSF B cells in virus-associated neuroinflammatory diseases"

Article Title: Immunophenotypic characterization of CSF B cells in virus-associated neuroinflammatory diseases

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1007042

Involvement of CD4 + CD25 + T cells with B cell help in CSF of HAM/TSP patients. (A) Comparison of HTLV-1 PVL in CSF of ACs (n = 7) and HAM/TSP patients (n = 36) using Mann-Whitney Test. (B) Correlation of HTLV-1 PVL with CD4 + CD25 + T cells and memory Tfh cells in HTLV-1-infected subjects using Spearman’s rank correlation test. (C) Comparison of IL-21 in CSF of HAM/TSP patients and ACs using unpaired t test. (D) Representative dot plots of IL-21 and Tax staining in CD4 + CD25 + T cells of a ND and a HAM/TSP patient after culture for 24 hours without any exogenous stimulation. (E) Detection of IL-21 in Tax-expressing CD4 + CD25 + T cells of HAM/TSP patients after culture for 24 hours without any exogenous stimulation. (F) Generation of ASCs subsets in B cells cultured with and without rhIL-21. Th e data were obtained from cultured B cells of NDs and HAM/TSP patients (n = 6). The horizontal line represents the mean. (G) Detection of human IgG in the B cell culture supernatants of NDs and HAM/TSP patients. (H) Detection of antibodies for HTLV-1 Gag and Tax in the B cell culture supernatants of HAM/TSP patients.
Figure Legend Snippet: Involvement of CD4 + CD25 + T cells with B cell help in CSF of HAM/TSP patients. (A) Comparison of HTLV-1 PVL in CSF of ACs (n = 7) and HAM/TSP patients (n = 36) using Mann-Whitney Test. (B) Correlation of HTLV-1 PVL with CD4 + CD25 + T cells and memory Tfh cells in HTLV-1-infected subjects using Spearman’s rank correlation test. (C) Comparison of IL-21 in CSF of HAM/TSP patients and ACs using unpaired t test. (D) Representative dot plots of IL-21 and Tax staining in CD4 + CD25 + T cells of a ND and a HAM/TSP patient after culture for 24 hours without any exogenous stimulation. (E) Detection of IL-21 in Tax-expressing CD4 + CD25 + T cells of HAM/TSP patients after culture for 24 hours without any exogenous stimulation. (F) Generation of ASCs subsets in B cells cultured with and without rhIL-21. Th e data were obtained from cultured B cells of NDs and HAM/TSP patients (n = 6). The horizontal line represents the mean. (G) Detection of human IgG in the B cell culture supernatants of NDs and HAM/TSP patients. (H) Detection of antibodies for HTLV-1 Gag and Tax in the B cell culture supernatants of HAM/TSP patients.

Techniques Used: MANN-WHITNEY, Infection, Staining, Expressing, Cell Culture

22) Product Images from "Antibody-Dependent Cellular Cytotoxicity-Competent Antibodies against HIV-1-Infected Cells in Plasma from HIV-Infected Subjects"

Article Title: Antibody-Dependent Cellular Cytotoxicity-Competent Antibodies against HIV-1-Infected Cells in Plasma from HIV-Infected Subjects

Journal: mBio

doi: 10.1128/mBio.02690-19

ADCC activity of effector PBMCs depends on the frequency of NK cells and CD16 levels on NK cells. Frequencies of NK cells (defined as live CD56 + CD3 − ) and levels of CD16 mean fluorescence intensity (MFI) on NK cells were evaluated in PBMCs from 9 HIV − donors tested in parallel for their capacity to mediate ADCC of coated CEM cells (cCEM) and uncoated CEM (unCEM) cells opsonized with HIV + IgG (as described for Fig. 1 ). (A nd B) Between-subject variation in the frequency of AnV + (% ADCC) cCEM (black bars) and unCEM (gray bars) cells generated in the ADCC-AnV assay using PBMCs and 1.5 μg/ml (A) or 15 μg/ml (B) of HIV + IgG to opsonize target cells. Data represent results from one experiment representative of three. Error bars indicate the standard deviations (SD) of results from replicates for each donor tested. (C and D) Spearman correlation between the frequency of NK cells present in live PBMC (% CD56 + CD3 − cells in live PBMC) and the frequency of AnV + (% ADCC) cCEM cells opsonized with 1.5 μg/ml (C) or 15 μg/ml (D) of HIV + IgG. (E and F) Spearman correlation between the MFI of CD16 expression on NK cells (MFI CD16 on CD56 + CD3 − cells) and the frequency of AnV + (% ADCC) cCEM cells opsonized with 1.5 μg/ml (E) or 15 μg/ml (F) of HIV + IgG.
Figure Legend Snippet: ADCC activity of effector PBMCs depends on the frequency of NK cells and CD16 levels on NK cells. Frequencies of NK cells (defined as live CD56 + CD3 − ) and levels of CD16 mean fluorescence intensity (MFI) on NK cells were evaluated in PBMCs from 9 HIV − donors tested in parallel for their capacity to mediate ADCC of coated CEM cells (cCEM) and uncoated CEM (unCEM) cells opsonized with HIV + IgG (as described for Fig. 1 ). (A nd B) Between-subject variation in the frequency of AnV + (% ADCC) cCEM (black bars) and unCEM (gray bars) cells generated in the ADCC-AnV assay using PBMCs and 1.5 μg/ml (A) or 15 μg/ml (B) of HIV + IgG to opsonize target cells. Data represent results from one experiment representative of three. Error bars indicate the standard deviations (SD) of results from replicates for each donor tested. (C and D) Spearman correlation between the frequency of NK cells present in live PBMC (% CD56 + CD3 − cells in live PBMC) and the frequency of AnV + (% ADCC) cCEM cells opsonized with 1.5 μg/ml (C) or 15 μg/ml (D) of HIV + IgG. (E and F) Spearman correlation between the MFI of CD16 expression on NK cells (MFI CD16 on CD56 + CD3 − cells) and the frequency of AnV + (% ADCC) cCEM cells opsonized with 1.5 μg/ml (E) or 15 μg/ml (F) of HIV + IgG.

Techniques Used: Activity Assay, Fluorescence, Generated, Expressing

Binding to cCEM and siCEM and ADCC activity mediated by anti-Env monoclonal and polyclonal Abs. siCEM cells and cCEM target cells were separately labeled with CFSE, opsonized with 15 μg/ml of each anti-Env Abs used as described for Fig. 3 . (A) Opsonized siCEM cells and cCEM cells were combined 1:1 and stained with anti-HSA ( y axis) to differentiate siCEM cells (CFSE + HSA + ) from cCEM cells (CFSE + HSA − ), and anti-human IgG Fc-specific secondary Ab was used to detect anti-Env binding ( x axis). The primary anti-Env Ab used for staining is identified above each density plot. Frequencies of HSA + and/or Env + cells are indicated in each quadrant. (B and C) Opsonized siCEM cells and cCEM cells were incubated side by side with isolated NK effector cells for 1 h. The y axes show ADCC activity (% ADCC) mediated by each of the anti-Env-specific MAbs (identified below each bar) measured as the frequencies of AnV + siCEM cells (B) and cCEM cells (C). Data represent averages ± SD of results from three independent experiments. Each dot represents a single NK cell donor. Significance was determined by comparing the percentages of ADCC between the anti-Env Abs used with HIV − IgG (*, P
Figure Legend Snippet: Binding to cCEM and siCEM and ADCC activity mediated by anti-Env monoclonal and polyclonal Abs. siCEM cells and cCEM target cells were separately labeled with CFSE, opsonized with 15 μg/ml of each anti-Env Abs used as described for Fig. 3 . (A) Opsonized siCEM cells and cCEM cells were combined 1:1 and stained with anti-HSA ( y axis) to differentiate siCEM cells (CFSE + HSA + ) from cCEM cells (CFSE + HSA − ), and anti-human IgG Fc-specific secondary Ab was used to detect anti-Env binding ( x axis). The primary anti-Env Ab used for staining is identified above each density plot. Frequencies of HSA + and/or Env + cells are indicated in each quadrant. (B and C) Opsonized siCEM cells and cCEM cells were incubated side by side with isolated NK effector cells for 1 h. The y axes show ADCC activity (% ADCC) mediated by each of the anti-Env-specific MAbs (identified below each bar) measured as the frequencies of AnV + siCEM cells (B) and cCEM cells (C). Data represent averages ± SD of results from three independent experiments. Each dot represents a single NK cell donor. Significance was determined by comparing the percentages of ADCC between the anti-Env Abs used with HIV − IgG (*, P

Techniques Used: Binding Assay, Activity Assay, Labeling, Staining, Incubation, Isolation

Inhibition of the ADCC-AnV activity of HIV + IgG or HIV + plasma samples using Fab fragments prepared from CD4i-specific MAb A32. cCEM cells and siCEM target cells were separately labeled with CFSE and preincubated with 10 μg/ml of A32 Fab or left untreated. cCEM cells and siCEM cells were then opsonized with A32 MAb or HIV + IgG or HIV + plasma samples and used as target cells in the ADCC-AnV assay. (A) Frequencies of AnV + cells (% ADCC) among CFSE + cCEM cells (left panel) and CFSE + siCEM cells (right panel) induced by ADCC following opsonization with A32 Fab alone or 10 μg/ml of A32 MAb to target cells preincubated or not with A32 Fab fragments. Data represent averages ± SD of results from two NK cell donors. This experiment was repeated three times. (B) Frequencies of AnV + results (% ADCC) in CFSE + cCEM cells (left panel) and CFSE + siCEM cells (right panel) induced by ADCC following opsonization of target cells by treatment with 0.37, 1.11, 3.33, and 10 μg/ml of HIV + IgG preincubated with 10 μg/ml of A32 Fab or left untreated. Error bars indicate SD of results from replicates, and significance was determined by comparing the percentages of ADCC with and without Fab for each opsonizing HIV + IgG concentration (***, P
Figure Legend Snippet: Inhibition of the ADCC-AnV activity of HIV + IgG or HIV + plasma samples using Fab fragments prepared from CD4i-specific MAb A32. cCEM cells and siCEM target cells were separately labeled with CFSE and preincubated with 10 μg/ml of A32 Fab or left untreated. cCEM cells and siCEM cells were then opsonized with A32 MAb or HIV + IgG or HIV + plasma samples and used as target cells in the ADCC-AnV assay. (A) Frequencies of AnV + cells (% ADCC) among CFSE + cCEM cells (left panel) and CFSE + siCEM cells (right panel) induced by ADCC following opsonization with A32 Fab alone or 10 μg/ml of A32 MAb to target cells preincubated or not with A32 Fab fragments. Data represent averages ± SD of results from two NK cell donors. This experiment was repeated three times. (B) Frequencies of AnV + results (% ADCC) in CFSE + cCEM cells (left panel) and CFSE + siCEM cells (right panel) induced by ADCC following opsonization of target cells by treatment with 0.37, 1.11, 3.33, and 10 μg/ml of HIV + IgG preincubated with 10 μg/ml of A32 Fab or left untreated. Error bars indicate SD of results from replicates, and significance was determined by comparing the percentages of ADCC with and without Fab for each opsonizing HIV + IgG concentration (***, P

Techniques Used: Inhibition, Activity Assay, Labeling, Concentration Assay

Characterization of the ADCC-AnV assay using HIV + IgG Ab-opsonized siCEM cells as target cells. (A) siCEM target cells were labeled with CFSE, opsonized with increasing doses of HIV + IgG (filled symbols) or HIV − IgG (empty symbols), and used as target cells in an ADCC-AnV assay with NK cells as effector cells. Data represent averages ± SD of results from two donors of NK cells, and significance was determined by comparing the frequencies of AnV + siCEM cells (%ADCC) between HIV + IgG and HIV − IgG for all IgG concentrations after background (No Ab) subtraction (***, P
Figure Legend Snippet: Characterization of the ADCC-AnV assay using HIV + IgG Ab-opsonized siCEM cells as target cells. (A) siCEM target cells were labeled with CFSE, opsonized with increasing doses of HIV + IgG (filled symbols) or HIV − IgG (empty symbols), and used as target cells in an ADCC-AnV assay with NK cells as effector cells. Data represent averages ± SD of results from two donors of NK cells, and significance was determined by comparing the frequencies of AnV + siCEM cells (%ADCC) between HIV + IgG and HIV − IgG for all IgG concentrations after background (No Ab) subtraction (***, P

Techniques Used: Labeling

Anti-Env Abs in HIV + plasma samples preferentially support ADCC of cCEM cells over siCEM cells. siCEM cells labeled with CFSE and PKH26 were combined 1:1 with cCEM cells labeled with CFSE only before opsonization with 10 individual HIV + plasma samples and were cocultured with NK effector cells. The y axes show percent ADCC as measured by the superimposed frequencies of AnV + siCEM cells (CFSE + PKH26 + ; black histograms) and cCEM cells (CFSE + PKH26 − ; gray histograms) with 15 μg/ml (A) and 1.5 μg/ml (B) of total IgG from each plasma sample used to opsonize target cells. Error bars indicate SD of results from replicates, and significance was determined by comparing the percentages of ADCC between siCEM cells and cCEM cells for each individual plasma sample (***, P
Figure Legend Snippet: Anti-Env Abs in HIV + plasma samples preferentially support ADCC of cCEM cells over siCEM cells. siCEM cells labeled with CFSE and PKH26 were combined 1:1 with cCEM cells labeled with CFSE only before opsonization with 10 individual HIV + plasma samples and were cocultured with NK effector cells. The y axes show percent ADCC as measured by the superimposed frequencies of AnV + siCEM cells (CFSE + PKH26 + ; black histograms) and cCEM cells (CFSE + PKH26 − ; gray histograms) with 15 μg/ml (A) and 1.5 μg/ml (B) of total IgG from each plasma sample used to opsonize target cells. Error bars indicate SD of results from replicates, and significance was determined by comparing the percentages of ADCC between siCEM cells and cCEM cells for each individual plasma sample (***, P

Techniques Used: Labeling

Flow-based measurement of ADCC activity using annexin V (AnV) staining of target cells. Uncoated CEM cells (unCEM) labeled with CFSE were mixed 1:1 with rgp120-coated CEM cells (cCEM) labeled with CFSE and PKH26 prior to the addition of opsonizing antibodies to be used as target (T) cells. Peripheral blood mononuclear cells (PBMCs) were used as effector (E) cells and mixed at an E:T ratio of 30:1. After 1 h of incubation, cells were stained with annexin V (AnV) and Live/Dead (LD) reagents to quantify the frequency of early/late apoptotic (AnV + ) and dead (LD + ) target CEM cells, respectively, by flow cytometry. (A) Gating strategy. Combined PBMC, cCEM cells, and unCEM cells were gated on by f orward sc atter A (FSC-A) and s ide sc atter A (SSC-A). The frequencies (%) of AnV + and LD + CEM cells were evaluated among cCEM cells (CFSE + PKH26 + , upper right panel) and unCEM cells (CFSE + PKH26 − ; lower right panel) cells opsonized with either 1.5 μg/ml (middle) or 15 μg/ml (right) of HIV + IgG compared to no antibody (No Ig, left). Percentages of AnV + and/or LD + cells are indicated in each quadrant. (B) Dose-response curves showing ADCC activity (% ADCC) as measured by the percentage of AnV + (black symbols) or LD + (gray symbols) cCEM target cells opsonized with either HIV + IgG (filled symbols) or HIV − IgG (empty symbols) after background (No Ab) subtraction. Error bars indicate the standard deviation (s.d.) of replicates and significance was determined by comparing the percentages of ADCC between HIV + IgG and HIV − IgG for all IgG concentrations using AnV or LD (*, P
Figure Legend Snippet: Flow-based measurement of ADCC activity using annexin V (AnV) staining of target cells. Uncoated CEM cells (unCEM) labeled with CFSE were mixed 1:1 with rgp120-coated CEM cells (cCEM) labeled with CFSE and PKH26 prior to the addition of opsonizing antibodies to be used as target (T) cells. Peripheral blood mononuclear cells (PBMCs) were used as effector (E) cells and mixed at an E:T ratio of 30:1. After 1 h of incubation, cells were stained with annexin V (AnV) and Live/Dead (LD) reagents to quantify the frequency of early/late apoptotic (AnV + ) and dead (LD + ) target CEM cells, respectively, by flow cytometry. (A) Gating strategy. Combined PBMC, cCEM cells, and unCEM cells were gated on by f orward sc atter A (FSC-A) and s ide sc atter A (SSC-A). The frequencies (%) of AnV + and LD + CEM cells were evaluated among cCEM cells (CFSE + PKH26 + , upper right panel) and unCEM cells (CFSE + PKH26 − ; lower right panel) cells opsonized with either 1.5 μg/ml (middle) or 15 μg/ml (right) of HIV + IgG compared to no antibody (No Ig, left). Percentages of AnV + and/or LD + cells are indicated in each quadrant. (B) Dose-response curves showing ADCC activity (% ADCC) as measured by the percentage of AnV + (black symbols) or LD + (gray symbols) cCEM target cells opsonized with either HIV + IgG (filled symbols) or HIV − IgG (empty symbols) after background (No Ab) subtraction. Error bars indicate the standard deviation (s.d.) of replicates and significance was determined by comparing the percentages of ADCC between HIV + IgG and HIV − IgG for all IgG concentrations using AnV or LD (*, P

Techniques Used: Flow Cytometry, Activity Assay, Staining, Labeling, Incubation, Cytometry, Standard Deviation

ADCC activity mediated by anti-Env monoclonal and polyclonal Abs to niCEM cells, byCEM cells, and unCEM cells (A to C) Four days p.i., infected CEM cells were labeled with CFSE, LD, and PKH26 and combined with uninfected CEM (unCEM) cells labeled with CFSE only as described for Fig. 3 . The combined CEM cells were opsonized with 15 μg/ml of each anti-Env Abs used as described for Fig. 3 and incubated with effector NK cells for 1 h followed by staining with anti-HSA and AnV. ADCC activity (% ADCC) was measured as the frequency of AnV + cells among niCEM (LD − CFSE + PKH26 + HSA + , red bars; A), byCEM (LD − CFSE + PKH26 + HSA − , blue bars; B) and unCEM (CFSE + PKH26 − , orange bars; C) cells. Data represent averages ± SD of results from three independent experiments. Each dot represents a single NK cell donor. Significance was determined by comparing the percentages of ADCC between each anti-Env Ab used and HIV − IgG (*, P
Figure Legend Snippet: ADCC activity mediated by anti-Env monoclonal and polyclonal Abs to niCEM cells, byCEM cells, and unCEM cells (A to C) Four days p.i., infected CEM cells were labeled with CFSE, LD, and PKH26 and combined with uninfected CEM (unCEM) cells labeled with CFSE only as described for Fig. 3 . The combined CEM cells were opsonized with 15 μg/ml of each anti-Env Abs used as described for Fig. 3 and incubated with effector NK cells for 1 h followed by staining with anti-HSA and AnV. ADCC activity (% ADCC) was measured as the frequency of AnV + cells among niCEM (LD − CFSE + PKH26 + HSA + , red bars; A), byCEM (LD − CFSE + PKH26 + HSA − , blue bars; B) and unCEM (CFSE + PKH26 − , orange bars; C) cells. Data represent averages ± SD of results from three independent experiments. Each dot represents a single NK cell donor. Significance was determined by comparing the percentages of ADCC between each anti-Env Ab used and HIV − IgG (*, P

Techniques Used: Activity Assay, Infection, Labeling, Incubation, Staining

23) Product Images from "Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls"

Article Title: Estradiol Affects Epstein–Barr Virus Reactivation-Induced Thyrotropin Receptor Antibody and Immunoglobulin Production in Graves’ Disease Patients and Healthy Controls

Journal: Viral Immunology

doi: 10.1089/vim.2018.0032

The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.
Figure Legend Snippet: The effect of estradiol on the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls and patients. There were no significant differences between healthy controls and patients for each concentration of estradiol.

Techniques Used: Concentration Assay

Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).
Figure Legend Snippet: Estradiol increased the ratio of IgG to IgG and IgM (IgG/IgG + IgM) in patients. Pileup production of Ig in healthy controls (a) and patients (b) . The ratio of IgG to IgG and IgM (IgG/IgG + IgM) in healthy controls (c) and patients (d) . In patients, the IgG/IgG + IgM ratio increased between 0 and 1 nM estradiol ( p = 0.063) and between 0 and 100 nM estradiol ( p = 0.128).

Techniques Used:

24) Product Images from "Circulating TFH Subset Distribution Is Strongly Affected in Lupus Patients with an Active Disease"

Article Title: Circulating TFH Subset Distribution Is Strongly Affected in Lupus Patients with an Active Disease

Journal: PLoS ONE

doi: 10.1371/journal.pone.0075319

IgE levels are increased in the serum of SLE patients. The quantification of IgG and IgE levels in the serum of inactive SLE patients (n = 17), active SLE patients (n = 6) and sex and age-matched healthy controls (n = 21) were determined by a sandwich ELISA assay. Results are expressed as the mean IgG concentration (g/L; A) or IgE concentration (ng/ml; B) from duplicate wells and correlation between IgG levels and T FH 2 cell frequency is represented (A). Each data point represents an individual subject and horizontal lines show the mean ± sem. * p
Figure Legend Snippet: IgE levels are increased in the serum of SLE patients. The quantification of IgG and IgE levels in the serum of inactive SLE patients (n = 17), active SLE patients (n = 6) and sex and age-matched healthy controls (n = 21) were determined by a sandwich ELISA assay. Results are expressed as the mean IgG concentration (g/L; A) or IgE concentration (ng/ml; B) from duplicate wells and correlation between IgG levels and T FH 2 cell frequency is represented (A). Each data point represents an individual subject and horizontal lines show the mean ± sem. * p

Techniques Used: Sandwich ELISA, Concentration Assay

25) Product Images from "Protein structure shapes immunodominance in the CD4 T cell response to yellow fever vaccination"

Article Title: Protein structure shapes immunodominance in the CD4 T cell response to yellow fever vaccination

Journal: Scientific Reports

doi: 10.1038/s41598-017-09331-w

CXCR5 + and CXCR5 − CD4 T cell response to YF virus C, prM and E peptides. ( a ) CD4 + CD45Ra − memory T cells from PBMC of healthy donors were sorted into CXCR5 + (upper right) and CXCR5 − cells (lower left). ( b ) IgG concentration revealed by ELISA in supernatants of sorted autologous B cells cultured for 10 days with either CXCR5 + or CXCR5 − cells in 4 independent experiments. Statistical signifiance was determined with the two-way ANOVA. ( c ) Percentage of cytokine events in sorted CXCR5 + and CXCR5 − subsets contributed by C, prM and E peptides as determined by intracellular cytokine staining. ( d ) Ratios of results with E and C peptides obtained in individual donors. Statistical signifiance was determined with the Wilcoxon matched-pairs signed rank test.
Figure Legend Snippet: CXCR5 + and CXCR5 − CD4 T cell response to YF virus C, prM and E peptides. ( a ) CD4 + CD45Ra − memory T cells from PBMC of healthy donors were sorted into CXCR5 + (upper right) and CXCR5 − cells (lower left). ( b ) IgG concentration revealed by ELISA in supernatants of sorted autologous B cells cultured for 10 days with either CXCR5 + or CXCR5 − cells in 4 independent experiments. Statistical signifiance was determined with the two-way ANOVA. ( c ) Percentage of cytokine events in sorted CXCR5 + and CXCR5 − subsets contributed by C, prM and E peptides as determined by intracellular cytokine staining. ( d ) Ratios of results with E and C peptides obtained in individual donors. Statistical signifiance was determined with the Wilcoxon matched-pairs signed rank test.

Techniques Used: Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Staining

26) Product Images from "Janus Kinase Inhibitor Baricitinib Modulates Human Innate and Adaptive Immune System"

Article Title: Janus Kinase Inhibitor Baricitinib Modulates Human Innate and Adaptive Immune System

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.01510

Baricitinib inhibits B cell differentiation. Human B cells were purified and cultured for 5 days in the presence of anti-human IgM Ab F(ab′)2 fragments and under interferon (IFN)-α stimulation. Plasmablast (CD3 − CD19 + CD20 − CD27 hi CD38 hi ) differentiation was assessed by flow cytometry, and IgG Ab titers and interleukin (IL)-6 concentrations in culture supernatants were measured by ELISA Cytometric Bead Array. (A) Percentage of plasmablasts. (B) Representative flow cytometry plots showing plasmablasts. (C) Percentage of plasmablast, IL-6, and IgG concentrations in the presence of baricitinib and other inhibitors. (D) Representative histogram data of annexin V/propidium iodide staining. (E) Rate of viable cells (annexin V neg /propidium iodide neg ). Data are mean ± SD of three different donors per group. * p
Figure Legend Snippet: Baricitinib inhibits B cell differentiation. Human B cells were purified and cultured for 5 days in the presence of anti-human IgM Ab F(ab′)2 fragments and under interferon (IFN)-α stimulation. Plasmablast (CD3 − CD19 + CD20 − CD27 hi CD38 hi ) differentiation was assessed by flow cytometry, and IgG Ab titers and interleukin (IL)-6 concentrations in culture supernatants were measured by ELISA Cytometric Bead Array. (A) Percentage of plasmablasts. (B) Representative flow cytometry plots showing plasmablasts. (C) Percentage of plasmablast, IL-6, and IgG concentrations in the presence of baricitinib and other inhibitors. (D) Representative histogram data of annexin V/propidium iodide staining. (E) Rate of viable cells (annexin V neg /propidium iodide neg ). Data are mean ± SD of three different donors per group. * p

Techniques Used: Cell Differentiation, Purification, Cell Culture, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Staining

27) Product Images from "Human Vγ9Vδ2-T Cells Synergize CD4+ T Follicular Helper Cells to Produce Influenza Virus-Specific Antibody"

Article Title: Human Vγ9Vδ2-T Cells Synergize CD4+ T Follicular Helper Cells to Produce Influenza Virus-Specific Antibody

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.00599

Vγ9Vδ2-T cells increased H9N2 virus-specific IgG and IgM productions in vivo . Specific immune cells-depleted and whole peripheral blood mononuclear cells (PBMC)-humanized mice were vaccinated with UV-inactivated H9N2 virus through i.p. pathway on day 0 and 7. Serum was collected on day 21 post the first vaccination. (A,B) Total IgG and IgM in serum (C,D) H9N2 virus-specific IgG and IgM in serum that treated with receptor destroyed enzyme (RDE) were determined by enzyme-linked immunosorbent assay. (E) H9N2-specific antibodies in serum that treated with RDE were determined by hemagglutination inhibition assay. The data shown were the represent of four independent experiments. (F) Representative histological analysis, immunohistological stainings for human CD4 T, B, and γδ-T cells in formalin-fixed paraffin-embedded spleen in humanized mice constructed with whole PBMCs after UV-H9N2 virus immunization. Scale bars = 100 μm. Bottom panels showed the higher-magnification views of the respective boxed areas in the top panels. Scale bars = 50 μm. The data shown are the mean ± SEM. * p
Figure Legend Snippet: Vγ9Vδ2-T cells increased H9N2 virus-specific IgG and IgM productions in vivo . Specific immune cells-depleted and whole peripheral blood mononuclear cells (PBMC)-humanized mice were vaccinated with UV-inactivated H9N2 virus through i.p. pathway on day 0 and 7. Serum was collected on day 21 post the first vaccination. (A,B) Total IgG and IgM in serum (C,D) H9N2 virus-specific IgG and IgM in serum that treated with receptor destroyed enzyme (RDE) were determined by enzyme-linked immunosorbent assay. (E) H9N2-specific antibodies in serum that treated with RDE were determined by hemagglutination inhibition assay. The data shown were the represent of four independent experiments. (F) Representative histological analysis, immunohistological stainings for human CD4 T, B, and γδ-T cells in formalin-fixed paraffin-embedded spleen in humanized mice constructed with whole PBMCs after UV-H9N2 virus immunization. Scale bars = 100 μm. Bottom panels showed the higher-magnification views of the respective boxed areas in the top panels. Scale bars = 50 μm. The data shown are the mean ± SEM. * p

Techniques Used: In Vivo, Mouse Assay, Enzyme-linked Immunosorbent Assay, HI Assay, Formalin-fixed Paraffin-Embedded, Construct

Vγ9Vδ2-T cells facilitated influenza virus-specific antibody production. Naïve CD4 T cells were cultured with UV-H9N2 virus-pulsed dendritic cells for 4–5 days, then, the activated CD4 T cells were cultured with CD19 + B cells from same donor with or without Vγ9Vδ2-T cells for another 7–10 days. (A,B) Total IgG and IgM in supernatant on day 7 were detected by enzyme-linked immunosorbent assay (ELISA). (C,D) H9N2 virus-specific IgG and IgM were detected by ELISA on day 7. Each dot means one donor. The data shown are the mean ± SEM. * p
Figure Legend Snippet: Vγ9Vδ2-T cells facilitated influenza virus-specific antibody production. Naïve CD4 T cells were cultured with UV-H9N2 virus-pulsed dendritic cells for 4–5 days, then, the activated CD4 T cells were cultured with CD19 + B cells from same donor with or without Vγ9Vδ2-T cells for another 7–10 days. (A,B) Total IgG and IgM in supernatant on day 7 were detected by enzyme-linked immunosorbent assay (ELISA). (C,D) H9N2 virus-specific IgG and IgM were detected by ELISA on day 7. Each dot means one donor. The data shown are the mean ± SEM. * p

Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay

28) Product Images from "Circulating TFH Subset Distribution Is Strongly Affected in Lupus Patients with an Active Disease"

Article Title: Circulating TFH Subset Distribution Is Strongly Affected in Lupus Patients with an Active Disease

Journal: PLoS ONE

doi: 10.1371/journal.pone.0075319

The frequency of memory B cells expressing IL-21R is enhanced in SLE patients and correlates with T FH 2 cell increase. IL-21 concentration was measured in the serum of SLE patients (n = 88) and healthy controls (n = 44) by a sandwich ELISA assay (A). Results are expressed as the mean IL-21 concentration (pg/ml) from duplicate wells. Each data point represents an individual subject and horizontal lines show the mean ± sem. Circulating B cell subsets were defined according to the expression of IgD and CD27 on CD19 + IL-21R + cells allowing the definition of 5 populations: Ab-secreting cells (ASC), non switched memory cells (NSmem), switched memory cells (Smem) and double negative memory cells (DNmem). Representative dot plots obtained with samples from one healthy control and one SLE patient are shown as an example (B). The distribution of each B cell subset among IL-21R + cells in healthy controls (n = 14) and SLE patients (n = 14) is represented (C). The frequency of CD27 + memory (left) and DN memory (right) B cells expressing IL-21R is compared between healthy controls (n = 14) and SLE patients (n = 14). Correlation between the IL-21R + DN memory cells and T FH 2 cell subset frequencies in SLE patients (n= 14) is represented (E). Each data point represents an individual subject; horizontal lines show the mean ± sem. * p
Figure Legend Snippet: The frequency of memory B cells expressing IL-21R is enhanced in SLE patients and correlates with T FH 2 cell increase. IL-21 concentration was measured in the serum of SLE patients (n = 88) and healthy controls (n = 44) by a sandwich ELISA assay (A). Results are expressed as the mean IL-21 concentration (pg/ml) from duplicate wells. Each data point represents an individual subject and horizontal lines show the mean ± sem. Circulating B cell subsets were defined according to the expression of IgD and CD27 on CD19 + IL-21R + cells allowing the definition of 5 populations: Ab-secreting cells (ASC), non switched memory cells (NSmem), switched memory cells (Smem) and double negative memory cells (DNmem). Representative dot plots obtained with samples from one healthy control and one SLE patient are shown as an example (B). The distribution of each B cell subset among IL-21R + cells in healthy controls (n = 14) and SLE patients (n = 14) is represented (C). The frequency of CD27 + memory (left) and DN memory (right) B cells expressing IL-21R is compared between healthy controls (n = 14) and SLE patients (n = 14). Correlation between the IL-21R + DN memory cells and T FH 2 cell subset frequencies in SLE patients (n= 14) is represented (E). Each data point represents an individual subject; horizontal lines show the mean ± sem. * p

Techniques Used: Expressing, Concentration Assay, Sandwich ELISA

IgE levels are increased in the serum of SLE patients. The quantification of IgG and IgE levels in the serum of inactive SLE patients (n = 17), active SLE patients (n = 6) and sex and age-matched healthy controls (n = 21) were determined by a sandwich ELISA assay. Results are expressed as the mean IgG concentration (g/L; A) or IgE concentration (ng/ml; B) from duplicate wells and correlation between IgG levels and T FH 2 cell frequency is represented (A). Each data point represents an individual subject and horizontal lines show the mean ± sem. * p
Figure Legend Snippet: IgE levels are increased in the serum of SLE patients. The quantification of IgG and IgE levels in the serum of inactive SLE patients (n = 17), active SLE patients (n = 6) and sex and age-matched healthy controls (n = 21) were determined by a sandwich ELISA assay. Results are expressed as the mean IgG concentration (g/L; A) or IgE concentration (ng/ml; B) from duplicate wells and correlation between IgG levels and T FH 2 cell frequency is represented (A). Each data point represents an individual subject and horizontal lines show the mean ± sem. * p

Techniques Used: Sandwich ELISA, Concentration Assay

29) Product Images from "Thyrotropin Receptor Antibody (TRAb)-IgM Levels Are Markedly Higher Than TRAb-IgG Levels in Graves' Disease Patients and Controls, and TRAb-IgM Production Is Related to Epstein–Barr Virus Reactivation"

Article Title: Thyrotropin Receptor Antibody (TRAb)-IgM Levels Are Markedly Higher Than TRAb-IgG Levels in Graves' Disease Patients and Controls, and TRAb-IgM Production Is Related to Epstein–Barr Virus Reactivation

Journal: Viral Immunology

doi: 10.1089/vim.2016.0043

Difference in serum TRAb levels between Graves' disease patients and controls. (a) Serum TRAb-IgG levels are significantly higher in Graves' disease patients than in controls (healthy subjects). (b) Serum TRAb-IgM levels are also significantly higher in Graves' disease patients than in controls. (a) p
Figure Legend Snippet: Difference in serum TRAb levels between Graves' disease patients and controls. (a) Serum TRAb-IgG levels are significantly higher in Graves' disease patients than in controls (healthy subjects). (b) Serum TRAb-IgM levels are also significantly higher in Graves' disease patients than in controls. (a) p

Techniques Used:

Immunoglobulin isotype difference in serum total Ig and TRAb levels in Graves' disease patients. The ratio of IgG to IgM between total Ig and TRAb was contrastive. * p
Figure Legend Snippet: Immunoglobulin isotype difference in serum total Ig and TRAb levels in Graves' disease patients. The ratio of IgG to IgM between total Ig and TRAb was contrastive. * p

Techniques Used:

Difference in serum TRAb-IgM levels between the high EBV Ab group and others. Serum TRAb-IgM levels are significantly higher in the high EBV Ab group (EBV reactivation) than in others ( p = 0.030*, the two-sample t -test). High EBV Ab represents samples with a high antibody index in EA-IgG and VCA-IgG, suggesting EBV reactivation. Others are constituted samples, except for cases with high EBV Ab levels. Ab, antibody; EA, early antigen; EBV, Epstein–Barr virus; VCA, virus capsid antigen.
Figure Legend Snippet: Difference in serum TRAb-IgM levels between the high EBV Ab group and others. Serum TRAb-IgM levels are significantly higher in the high EBV Ab group (EBV reactivation) than in others ( p = 0.030*, the two-sample t -test). High EBV Ab represents samples with a high antibody index in EA-IgG and VCA-IgG, suggesting EBV reactivation. Others are constituted samples, except for cases with high EBV Ab levels. Ab, antibody; EA, early antigen; EBV, Epstein–Barr virus; VCA, virus capsid antigen.

Techniques Used:

30) Product Images from "Anti-α-enolase is a prognostic marker in postoperative lung cancer patients"

Article Title: Anti-α-enolase is a prognostic marker in postoperative lung cancer patients

Journal: Oncotarget

doi:

Levels of circulating anti-ENO1 Ab in NSCLC patients before and after tumor removal A. Plasma from healthy donors (white bars, n = 36) and NSCLC patients (black bars, n = 85) were collected and the levels of anti-ENO1 Ab and total IgG were quantified by ELISA. Bars represent mean ± SEM. Statistics were performed using Student's t test. B. Correlation between the expression of ENO1 in tumors and the level of anti-ENO1 Ab in plasma of patients before surgery. Statistics were performed using Pearson's Correlation Coefficient analysis. C. Plasma from the same patients were collected before and 1 month after surgery to remove the tumor. The level of anti-ENO1 Ab in the plasma was determined by ELISA. Each line indicates the change in the plasma level of anti-ENO1 Ab in the same patient ( n = 85) before and after surgery. Statistics were performed using one tailed paired t -test. * P
Figure Legend Snippet: Levels of circulating anti-ENO1 Ab in NSCLC patients before and after tumor removal A. Plasma from healthy donors (white bars, n = 36) and NSCLC patients (black bars, n = 85) were collected and the levels of anti-ENO1 Ab and total IgG were quantified by ELISA. Bars represent mean ± SEM. Statistics were performed using Student's t test. B. Correlation between the expression of ENO1 in tumors and the level of anti-ENO1 Ab in plasma of patients before surgery. Statistics were performed using Pearson's Correlation Coefficient analysis. C. Plasma from the same patients were collected before and 1 month after surgery to remove the tumor. The level of anti-ENO1 Ab in the plasma was determined by ELISA. Each line indicates the change in the plasma level of anti-ENO1 Ab in the same patient ( n = 85) before and after surgery. Statistics were performed using one tailed paired t -test. * P

Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, One-tailed Test

31) Product Images from "A novel neutralizing human monoclonal antibody broadly abrogates hepatitis C virus infection in vitro and in vivo"

Article Title: A novel neutralizing human monoclonal antibody broadly abrogates hepatitis C virus infection in vitro and in vivo

Journal: Antiviral research

doi: 10.1016/j.antiviral.2017.10.015

Main binding motif recognized by mAb 2A5 (A) Alignment of the E2-region spanning AA420-452 of the different HCV strains used in the binding and neutralization studies. Residues found critical for mAb 2A5-binding (AA433, AA438, AA441, AA442, AA443 and AA446) are highlighted and, for each strain, the simplified AA-motif is shown on the right. (B) A hierarchy in neutralization potential was deduced for the binding motifs. The AA-motif corresponding to the H77c strain is marked with a black box. AA-motifs shown on the left-side of ‘ > ‘ have better neutralization properties compared to AA-motifs shown on the right-side. Motifs from ‘difficult to neutralize’ strains HCV-S52, JC1 and J6 are highlighted in red boxes. Motifs from HCV-strains that can be efficiently neutralized by 2A5 (P12_1091, HK6a and UKN5.15.7) are indicated in green boxes. (*) represents i dentical motifs with different neutralization efficiencies.
Figure Legend Snippet: Main binding motif recognized by mAb 2A5 (A) Alignment of the E2-region spanning AA420-452 of the different HCV strains used in the binding and neutralization studies. Residues found critical for mAb 2A5-binding (AA433, AA438, AA441, AA442, AA443 and AA446) are highlighted and, for each strain, the simplified AA-motif is shown on the right. (B) A hierarchy in neutralization potential was deduced for the binding motifs. The AA-motif corresponding to the H77c strain is marked with a black box. AA-motifs shown on the left-side of ‘ > ‘ have better neutralization properties compared to AA-motifs shown on the right-side. Motifs from ‘difficult to neutralize’ strains HCV-S52, JC1 and J6 are highlighted in red boxes. Motifs from HCV-strains that can be efficiently neutralized by 2A5 (P12_1091, HK6a and UKN5.15.7) are indicated in green boxes. (*) represents i dentical motifs with different neutralization efficiencies.

Techniques Used: Binding Assay, Neutralization

Binding characteristics of mAbs 2A5 and AP33 (A) mAbs 2A5 and AP33 were serially diluted and incubated on plates pre-coated with cell lysates containing HCV E1E2 derived from prototype isolates (H77c, JFH1, S52) and gt1b patient-derived viral isolates (P09_VA, P12_VA, P12_1091). Dose-dependent binding is expressed as percentage of maximal binding (mean ± standard deviation (error bars)). (B) EC 50 values were calculated from dose-response curves shown in (A) . All conditions were performed in duplicate.
Figure Legend Snippet: Binding characteristics of mAbs 2A5 and AP33 (A) mAbs 2A5 and AP33 were serially diluted and incubated on plates pre-coated with cell lysates containing HCV E1E2 derived from prototype isolates (H77c, JFH1, S52) and gt1b patient-derived viral isolates (P09_VA, P12_VA, P12_1091). Dose-dependent binding is expressed as percentage of maximal binding (mean ± standard deviation (error bars)). (B) EC 50 values were calculated from dose-response curves shown in (A) . All conditions were performed in duplicate.

Techniques Used: Binding Assay, Incubation, Derivative Assay, Standard Deviation

Neutralization of HCVpp by mAbs 2A5 and AP33 (A) HCVpp expressing E1E2 derived from prototype isolates (H77c, JFH1, S52) and gt1b patient-derived viral isolates (P09_VA, P09_VB, P09_779, P12_VA and P12_1091) were incubated for 1 h at 37 °C with serial dilutions of mAbs 2A5 or AP33 and added to Hep3B cells. HCVpp entry was analyzed by luciferase reporter gene expression and normalized to isotype controls. Neutralization is expressed as % neutralization (mean ± standard deviation (error bars)). (B) IC 50 values were calculated from dose-response curves shown in (A) . All conditions were performed in quadruplicate.
Figure Legend Snippet: Neutralization of HCVpp by mAbs 2A5 and AP33 (A) HCVpp expressing E1E2 derived from prototype isolates (H77c, JFH1, S52) and gt1b patient-derived viral isolates (P09_VA, P09_VB, P09_779, P12_VA and P12_1091) were incubated for 1 h at 37 °C with serial dilutions of mAbs 2A5 or AP33 and added to Hep3B cells. HCVpp entry was analyzed by luciferase reporter gene expression and normalized to isotype controls. Neutralization is expressed as % neutralization (mean ± standard deviation (error bars)). (B) IC 50 values were calculated from dose-response curves shown in (A) . All conditions were performed in quadruplicate.

Techniques Used: Neutralization, Expressing, Derivative Assay, Incubation, Luciferase, Standard Deviation

Neutralization of HCVcc by mAbs 2A5 and AP33 (A) HCVcc expressing the structural proteins of genotype 1a, 1b, 2a, 3a, 4a, 5a, 6a and 7a isolates were pre-incubated with three-fold serial dilutions of mAbs 2A5, AP33 or a control Ab. The mixture was transferred to Huh7.5.RFP cells and incubated for 4 h before washing. Two days later, HCV-infected foci were visualized using an NS5A-specific antibody and counted. Results are expressed as percentage of infectivity (mean ± standard deviation (error bars)). (B) IC 50 values were calculated from dose-response curves shown in (A) . All conditions were performed in triplicate.
Figure Legend Snippet: Neutralization of HCVcc by mAbs 2A5 and AP33 (A) HCVcc expressing the structural proteins of genotype 1a, 1b, 2a, 3a, 4a, 5a, 6a and 7a isolates were pre-incubated with three-fold serial dilutions of mAbs 2A5, AP33 or a control Ab. The mixture was transferred to Huh7.5.RFP cells and incubated for 4 h before washing. Two days later, HCV-infected foci were visualized using an NS5A-specific antibody and counted. Results are expressed as percentage of infectivity (mean ± standard deviation (error bars)). (B) IC 50 values were calculated from dose-response curves shown in (A) . All conditions were performed in triplicate.

Techniques Used: Neutralization, Expressing, Incubation, Infection, Standard Deviation

mAb 2A5 protects human liver chimeric mice from HCV challenge Three days before viral infection, humanized mice were passively immunized with 1 mg of mAb 2A5. Mice were challenged with mH77 (gt1a, 10 4 IU HCV RNA), mP05 (gt1b, 10 4 IU), mED43 (gt4a, 10 4 IU) or mHK6a (gt6a, 10 5 IU). Viremia was quantified until week 8 post infection. 2A5-treated and control mice are represented by closed and open circles, respectively. Mice that scored HCV RNA negative are displayed below the dashed line representing the limit of quantification (LOQ; HCV RNA
Figure Legend Snippet: mAb 2A5 protects human liver chimeric mice from HCV challenge Three days before viral infection, humanized mice were passively immunized with 1 mg of mAb 2A5. Mice were challenged with mH77 (gt1a, 10 4 IU HCV RNA), mP05 (gt1b, 10 4 IU), mED43 (gt4a, 10 4 IU) or mHK6a (gt6a, 10 5 IU). Viremia was quantified until week 8 post infection. 2A5-treated and control mice are represented by closed and open circles, respectively. Mice that scored HCV RNA negative are displayed below the dashed line representing the limit of quantification (LOQ; HCV RNA

Techniques Used: Mouse Assay, Infection

Recognition by mAb 2A5 is conformation dependent (A) To reveal whether mAb 2A5 recognizes a linear or a conformational epitope, mAbs 2A5, AP33, HC84.26 or HC33.1 were incubated at 5 μg/mL with native as well as denatured full-length H77c E1E2 cell lysates. Bound antibodies were detected with HRP-conjugated secondary antibodies and the signal was normalized to that obtained with the native protein. (B) To analyze whether 2A5 competes with other mAb, a competition EIA was performed wherein anti-E2 (AP33, HC33.1, HC84.26, HC-1AM, CBH-7, 1:7 and 3/11) and anti-E1 (A4) mAbs were used as competing antibodies for binding to the E1E2 protein of the H77c isolate. Three-fold serial dilutions of competing mAbs were used starting at 30 μg/mL. The binding of biotinylated versions of mAb 2A5 (left panel) and AP33 (right panel) was measured using HRP-conjugated streptavidin. Mal1C is an irrelevant antibody that was included as negative control. All conditions were performed in duplicate.
Figure Legend Snippet: Recognition by mAb 2A5 is conformation dependent (A) To reveal whether mAb 2A5 recognizes a linear or a conformational epitope, mAbs 2A5, AP33, HC84.26 or HC33.1 were incubated at 5 μg/mL with native as well as denatured full-length H77c E1E2 cell lysates. Bound antibodies were detected with HRP-conjugated secondary antibodies and the signal was normalized to that obtained with the native protein. (B) To analyze whether 2A5 competes with other mAb, a competition EIA was performed wherein anti-E2 (AP33, HC33.1, HC84.26, HC-1AM, CBH-7, 1:7 and 3/11) and anti-E1 (A4) mAbs were used as competing antibodies for binding to the E1E2 protein of the H77c isolate. Three-fold serial dilutions of competing mAbs were used starting at 30 μg/mL. The binding of biotinylated versions of mAb 2A5 (left panel) and AP33 (right panel) was measured using HRP-conjugated streptavidin. Mal1C is an irrelevant antibody that was included as negative control. All conditions were performed in duplicate.

Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay, Binding Assay, Negative Control

Epitope mapping of mAb 2A5 The binding pattern of mAb 2A5 was analyzed using gt1a- and gt1b-derived sequences, of which the E2 region spanning amino acids (AA)433–443 of the different clones (cl) is shown (A) . After incubation with surface-attached peptides, bound antibodies were detected with an HRP-conjugated anti-human secondary antibody. The 2A5-binding pattern was analyzed using mAb (50 μg/mL) and peptides spanning AA-region 433–443, derived from naturally occurring gt1a (clone a-b) and gt1b (clone c-d) isolates (B) , and A-substituted peptides (L433A, Y443A) from gt1a_clone a and gt1b_clone d (C) . Binding was normalized to the best binding peptide (100% binding). (D) To elaborate on the length of the epitope, the efficiency of 2A5-binding was compared between peptides covering region AA433-443 and peptides including an additional upstream sequence (AA433-452), all derived from gt1a_clone a. (E) To further elaborate on 2A5-anchor residues within the region of interest (AA433-452), the binding efficiency was compared between peptides derived from the prototype isolate H77c and their A-substituted counterparts. The corresponding color-codes, representing the different A-substituted sequences, and the % binding relative to native H77c at 10 μg mAb/mL are shown in (F). (G) To confirm these results and to reveal additional contact residues for mAb binding, epitope mapping was done using E1E2 proteins of the H77c isolate with A-substitutions in 3 regions (AA419-447, AA522-536 and AA612-617). 2A5-binding to full-length native and mutated E1E2 cell lysates was analyzed. Results represent the % binding relative to E1E2_H77c at 10 μg mAb/mL (F, G) .
Figure Legend Snippet: Epitope mapping of mAb 2A5 The binding pattern of mAb 2A5 was analyzed using gt1a- and gt1b-derived sequences, of which the E2 region spanning amino acids (AA)433–443 of the different clones (cl) is shown (A) . After incubation with surface-attached peptides, bound antibodies were detected with an HRP-conjugated anti-human secondary antibody. The 2A5-binding pattern was analyzed using mAb (50 μg/mL) and peptides spanning AA-region 433–443, derived from naturally occurring gt1a (clone a-b) and gt1b (clone c-d) isolates (B) , and A-substituted peptides (L433A, Y443A) from gt1a_clone a and gt1b_clone d (C) . Binding was normalized to the best binding peptide (100% binding). (D) To elaborate on the length of the epitope, the efficiency of 2A5-binding was compared between peptides covering region AA433-443 and peptides including an additional upstream sequence (AA433-452), all derived from gt1a_clone a. (E) To further elaborate on 2A5-anchor residues within the region of interest (AA433-452), the binding efficiency was compared between peptides derived from the prototype isolate H77c and their A-substituted counterparts. The corresponding color-codes, representing the different A-substituted sequences, and the % binding relative to native H77c at 10 μg mAb/mL are shown in (F). (G) To confirm these results and to reveal additional contact residues for mAb binding, epitope mapping was done using E1E2 proteins of the H77c isolate with A-substitutions in 3 regions (AA419-447, AA522-536 and AA612-617). 2A5-binding to full-length native and mutated E1E2 cell lysates was analyzed. Results represent the % binding relative to E1E2_H77c at 10 μg mAb/mL (F, G) .

Techniques Used: Binding Assay, Derivative Assay, Incubation, Sequencing

32) Product Images from "A long non-coding SINEUP RNA boosts semi-stable production of fully human monoclonal antibodies in HEK293E cells"

Article Title: A long non-coding SINEUP RNA boosts semi-stable production of fully human monoclonal antibodies in HEK293E cells

Journal: mAbs

doi: 10.1080/19420862.2018.1463945

Increased production of anti-CLDN1 B9x IgG4 by transient and stable expression of the SINEUP -40+32 construct > . A. The ELISA assay shows increased production of full human B9x IgG4 in cells co-transfected with the construct expressing the SINEUP -40+32 lncRNA, targeting both heavy- and light-chain mRNAs of the anti-CLDN1 B9x antibody. B. The ELISA assay, revealing full human IgG s , shows the antibodies secreted by HEK293E (wt) and HEK293E_SINEUP -40+32 cells stably expressing the lncRNA (pool or isolated clones 1, 3, 12, 15) after 8 days following transfection with pEUVH8.2_B9x and pEUVL4.2_SA_B9x vectors. The asterisks indicate p values
Figure Legend Snippet: Increased production of anti-CLDN1 B9x IgG4 by transient and stable expression of the SINEUP -40+32 construct > . A. The ELISA assay shows increased production of full human B9x IgG4 in cells co-transfected with the construct expressing the SINEUP -40+32 lncRNA, targeting both heavy- and light-chain mRNAs of the anti-CLDN1 B9x antibody. B. The ELISA assay, revealing full human IgG s , shows the antibodies secreted by HEK293E (wt) and HEK293E_SINEUP -40+32 cells stably expressing the lncRNA (pool or isolated clones 1, 3, 12, 15) after 8 days following transfection with pEUVH8.2_B9x and pEUVL4.2_SA_B9x vectors. The asterisks indicate p values

Techniques Used: Expressing, Construct, Enzyme-linked Immunosorbent Assay, Transfection, Stable Transfection, Isolation, Clone Assay

33) Product Images from "Mechanistic insights into the impairment of memory B cells and antibody production in the elderly"

Article Title: Mechanistic insights into the impairment of memory B cells and antibody production in the elderly

Journal: Age

doi: 10.1007/s11357-011-9371-9

Kinetics of B cell responses before and at 1 and 9 months after two-dose primary vaccination and at 1 and 9 months after booster. a , b Geometric means with 95% CI of plasma TBEV-specific IgG concentrations (ELISA) and c , d neutralizing
Figure Legend Snippet: Kinetics of B cell responses before and at 1 and 9 months after two-dose primary vaccination and at 1 and 9 months after booster. a , b Geometric means with 95% CI of plasma TBEV-specific IgG concentrations (ELISA) and c , d neutralizing

Techniques Used: Enzyme-linked Immunosorbent Assay

34) Product Images from "Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia"

Article Title: Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia

Journal: PLoS ONE

doi: 10.1371/journal.pone.0162973

Contents of total proteins, immunoglobulin G (IgG), IgA, IgM, albumin, and complement 3 (C3) and C4 fractions throughout the purification process.
Figure Legend Snippet: Contents of total proteins, immunoglobulin G (IgG), IgA, IgM, albumin, and complement 3 (C3) and C4 fractions throughout the purification process.

Techniques Used: Purification

35) Product Images from "Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia"

Article Title: Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent Fetal and Neonatal Alloimmune Thrombocytopenia

Journal: PLoS ONE

doi: 10.1371/journal.pone.0162973

Contents of total proteins, immunoglobulin G (IgG), IgA, IgM, albumin, and complement 3 (C3) and C4 fractions throughout the purification process.
Figure Legend Snippet: Contents of total proteins, immunoglobulin G (IgG), IgA, IgM, albumin, and complement 3 (C3) and C4 fractions throughout the purification process.

Techniques Used: Purification

SDS-PAGE profile of anti-human platelet antigen (HPA)-1a fractions generated throughout the purification process. A: Non-reducing conditions; B: reducing conditions. 1: cryoprecipitate-poor plasma; 2: caprylic acid supernatant; 3: S HyperCel breakthrough; 4: S HyperCel eluate; 5: HyperCel STAR-AX breakthrough; 6: HyperCel STAR-AX eluate; 7: after nanofiltration; 8: after concentration; 9: final fraction after dialysis. The left lane shows molecular weight markers. Staining was with Coomassie blue. FL-IgG, intact whole IgG; HC-IgG, heavy-chain IgG; LC-IgG, light-chain IgG.
Figure Legend Snippet: SDS-PAGE profile of anti-human platelet antigen (HPA)-1a fractions generated throughout the purification process. A: Non-reducing conditions; B: reducing conditions. 1: cryoprecipitate-poor plasma; 2: caprylic acid supernatant; 3: S HyperCel breakthrough; 4: S HyperCel eluate; 5: HyperCel STAR-AX breakthrough; 6: HyperCel STAR-AX eluate; 7: after nanofiltration; 8: after concentration; 9: final fraction after dialysis. The left lane shows molecular weight markers. Staining was with Coomassie blue. FL-IgG, intact whole IgG; HC-IgG, heavy-chain IgG; LC-IgG, light-chain IgG.

Techniques Used: SDS Page, Generated, Purification, Concentration Assay, Molecular Weight, Staining

36) Product Images from "A long non-coding SINEUP RNA boosts semi-stable production of fully human monoclonal antibodies in HEK293E cells"

Article Title: A long non-coding SINEUP RNA boosts semi-stable production of fully human monoclonal antibodies in HEK293E cells

Journal: mAbs

doi: 10.1080/19420862.2018.1463945

Increased production of anti-CLDN1 B9x IgG4 by transient and stable expression of the SINEUP -40+32 construct > . A. The ELISA assay shows increased production of full human B9x IgG4 in cells co-transfected with the construct expressing the SINEUP -40+32 lncRNA, targeting both heavy- and light-chain mRNAs of the anti-CLDN1 B9x antibody. B. The ELISA assay, revealing full human IgG s , shows the antibodies secreted by HEK293E (wt) and HEK293E_SINEUP -40+32 cells stably expressing the lncRNA (pool or isolated clones 1, 3, 12, 15) after 8 days following transfection with pEUVH8.2_B9x and pEUVL4.2_SA_B9x vectors. The asterisks indicate p values
Figure Legend Snippet: Increased production of anti-CLDN1 B9x IgG4 by transient and stable expression of the SINEUP -40+32 construct > . A. The ELISA assay shows increased production of full human B9x IgG4 in cells co-transfected with the construct expressing the SINEUP -40+32 lncRNA, targeting both heavy- and light-chain mRNAs of the anti-CLDN1 B9x antibody. B. The ELISA assay, revealing full human IgG s , shows the antibodies secreted by HEK293E (wt) and HEK293E_SINEUP -40+32 cells stably expressing the lncRNA (pool or isolated clones 1, 3, 12, 15) after 8 days following transfection with pEUVH8.2_B9x and pEUVL4.2_SA_B9x vectors. The asterisks indicate p values

Techniques Used: Expressing, Construct, Enzyme-linked Immunosorbent Assay, Transfection, Stable Transfection, Isolation, Clone Assay

37) Product Images from "Anti-α-enolase is a prognostic marker in postoperative lung cancer patients"

Article Title: Anti-α-enolase is a prognostic marker in postoperative lung cancer patients

Journal: Oncotarget

doi:

Levels of circulating anti-ENO1 Ab in NSCLC patients before and after tumor removal A. Plasma from healthy donors (white bars, n = 36) and NSCLC patients (black bars, n = 85) were collected and the levels of anti-ENO1 Ab and total IgG were quantified by ELISA. Bars represent mean ± SEM. Statistics were performed using Student's t test. B. Correlation between the expression of ENO1 in tumors and the level of anti-ENO1 Ab in plasma of patients before surgery. Statistics were performed using Pearson's Correlation Coefficient analysis. C. Plasma from the same patients were collected before and 1 month after surgery to remove the tumor. The level of anti-ENO1 Ab in the plasma was determined by ELISA. Each line indicates the change in the plasma level of anti-ENO1 Ab in the same patient ( n = 85) before and after surgery. Statistics were performed using one tailed paired t -test. * P
Figure Legend Snippet: Levels of circulating anti-ENO1 Ab in NSCLC patients before and after tumor removal A. Plasma from healthy donors (white bars, n = 36) and NSCLC patients (black bars, n = 85) were collected and the levels of anti-ENO1 Ab and total IgG were quantified by ELISA. Bars represent mean ± SEM. Statistics were performed using Student's t test. B. Correlation between the expression of ENO1 in tumors and the level of anti-ENO1 Ab in plasma of patients before surgery. Statistics were performed using Pearson's Correlation Coefficient analysis. C. Plasma from the same patients were collected before and 1 month after surgery to remove the tumor. The level of anti-ENO1 Ab in the plasma was determined by ELISA. Each line indicates the change in the plasma level of anti-ENO1 Ab in the same patient ( n = 85) before and after surgery. Statistics were performed using one tailed paired t -test. * P

Techniques Used: Enzyme-linked Immunosorbent Assay, Expressing, One-tailed Test

38) Product Images from "A monoclonal antibody-based immunoassay to measure the antibody response against the repeat region of the circumsporozoite protein of Plasmodium falciparum"

Article Title: A monoclonal antibody-based immunoassay to measure the antibody response against the repeat region of the circumsporozoite protein of Plasmodium falciparum

Journal: Malaria Journal

doi: 10.1186/s12936-016-1596-8

Comparison of antibody concentrations in sera from different time points and treatment groups. In both studies sera were obtained after the second (PD2) and third (PD3) vaccine dose and antibody concentrations were measured with R32LR and MAL1C-competition ELISA. In study 1 results obtained in the subjects receiving three doses of RTS,S/AS01 (3xRTS,S/AS01) were compared with those generated in subjects vaccinated with one dose of Ad35.CS.01 followed by two doses of RTS,S/AS01. In study 2, subjects vaccinated with three standard doses of RTS,S/AS01 given with monthly intervals at month 0, 1 and 2 (012) were compared with subjects given a standard dose at months 0 and 1 and 1/5 th of the standard dose at month 5 (017). No significant differences were observed between the study groups or between 2 and 3 vaccine doses
Figure Legend Snippet: Comparison of antibody concentrations in sera from different time points and treatment groups. In both studies sera were obtained after the second (PD2) and third (PD3) vaccine dose and antibody concentrations were measured with R32LR and MAL1C-competition ELISA. In study 1 results obtained in the subjects receiving three doses of RTS,S/AS01 (3xRTS,S/AS01) were compared with those generated in subjects vaccinated with one dose of Ad35.CS.01 followed by two doses of RTS,S/AS01. In study 2, subjects vaccinated with three standard doses of RTS,S/AS01 given with monthly intervals at month 0, 1 and 2 (012) were compared with subjects given a standard dose at months 0 and 1 and 1/5 th of the standard dose at month 5 (017). No significant differences were observed between the study groups or between 2 and 3 vaccine doses

Techniques Used: Enzyme-linked Immunosorbent Assay, Generated

Correlation between R32LR ELISA and MAL1C-competition ELISA. Sera from participants in study 1 ( panel A ) and study 2 ( panel B ) taken after the second and third vaccine doses (immediately prior to mosquito bite challenge) were analysed with the standard R32LR ELISA and the MAL1C-competition ELISA. Results obtained with both assays are plotted and correlation is calculated with Pearson’s correlation
Figure Legend Snippet: Correlation between R32LR ELISA and MAL1C-competition ELISA. Sera from participants in study 1 ( panel A ) and study 2 ( panel B ) taken after the second and third vaccine doses (immediately prior to mosquito bite challenge) were analysed with the standard R32LR ELISA and the MAL1C-competition ELISA. Results obtained with both assays are plotted and correlation is calculated with Pearson’s correlation

Techniques Used: Enzyme-linked Immunosorbent Assay

Development of the MAL1C-competition ELISA. Optimal dilutions of B-MAL1C and streptavidin-HRP were defined using a checkerboard titration experiment ( a ). At a 1/16,000 dilution of streptavidin-HRP a sigmoidal curve was observed; this dilution was used for further experiments. B-MAL1C dilutions in the range between the maximal absorbance (plateau observed at 1/50,000) and half max (max/2 observed at 1/400,000) were explored in an inhibition set up using defined sera with high and low antibody content in R32LR ELISA assay ( b ) and the three monoclonal antibodies, MAL1C, MAL2A and MAL3B ( c ). The data shown in panels b and c were obtained with the highest serum concentrations (starting dilution 1/5) and with mAb concentrations of 5 µg/ml
Figure Legend Snippet: Development of the MAL1C-competition ELISA. Optimal dilutions of B-MAL1C and streptavidin-HRP were defined using a checkerboard titration experiment ( a ). At a 1/16,000 dilution of streptavidin-HRP a sigmoidal curve was observed; this dilution was used for further experiments. B-MAL1C dilutions in the range between the maximal absorbance (plateau observed at 1/50,000) and half max (max/2 observed at 1/400,000) were explored in an inhibition set up using defined sera with high and low antibody content in R32LR ELISA assay ( b ) and the three monoclonal antibodies, MAL1C, MAL2A and MAL3B ( c ). The data shown in panels b and c were obtained with the highest serum concentrations (starting dilution 1/5) and with mAb concentrations of 5 µg/ml

Techniques Used: Enzyme-linked Immunosorbent Assay, Titration, Inhibition

Correlation between protection from infection and antibody content, measured with R32LR and MAL1C-competition ELISA. Antibody concentrations in the sera obtained immediately before challenge with infected mosquito’s from participants at the study 1 (n = 46) and study 2 (n = 46) studies were measured with the R32LR ELISA and MAL1C-competition ELISA. Antibody levels in protected and non-protected vaccine recipients were compared and no significant differences were observed irrespective of the assay used
Figure Legend Snippet: Correlation between protection from infection and antibody content, measured with R32LR and MAL1C-competition ELISA. Antibody concentrations in the sera obtained immediately before challenge with infected mosquito’s from participants at the study 1 (n = 46) and study 2 (n = 46) studies were measured with the R32LR ELISA and MAL1C-competition ELISA. Antibody levels in protected and non-protected vaccine recipients were compared and no significant differences were observed irrespective of the assay used

Techniques Used: Infection, Enzyme-linked Immunosorbent Assay

39) Product Images from "Janus Kinase Inhibitor Baricitinib Modulates Human Innate and Adaptive Immune System"

Article Title: Janus Kinase Inhibitor Baricitinib Modulates Human Innate and Adaptive Immune System

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.01510

Baricitinib inhibits B cell differentiation. Human B cells were purified and cultured for 5 days in the presence of anti-human IgM Ab F(ab′)2 fragments and under interferon (IFN)-α stimulation. Plasmablast (CD3 − CD19 + CD20 − CD27 hi CD38 hi ) differentiation was assessed by flow cytometry, and IgG Ab titers and interleukin (IL)-6 concentrations in culture supernatants were measured by ELISA Cytometric Bead Array. (A) Percentage of plasmablasts. (B) Representative flow cytometry plots showing plasmablasts. (C) Percentage of plasmablast, IL-6, and IgG concentrations in the presence of baricitinib and other inhibitors. (D) Representative histogram data of annexin V/propidium iodide staining. (E) Rate of viable cells (annexin V neg /propidium iodide neg ). Data are mean ± SD of three different donors per group. * p
Figure Legend Snippet: Baricitinib inhibits B cell differentiation. Human B cells were purified and cultured for 5 days in the presence of anti-human IgM Ab F(ab′)2 fragments and under interferon (IFN)-α stimulation. Plasmablast (CD3 − CD19 + CD20 − CD27 hi CD38 hi ) differentiation was assessed by flow cytometry, and IgG Ab titers and interleukin (IL)-6 concentrations in culture supernatants were measured by ELISA Cytometric Bead Array. (A) Percentage of plasmablasts. (B) Representative flow cytometry plots showing plasmablasts. (C) Percentage of plasmablast, IL-6, and IgG concentrations in the presence of baricitinib and other inhibitors. (D) Representative histogram data of annexin V/propidium iodide staining. (E) Rate of viable cells (annexin V neg /propidium iodide neg ). Data are mean ± SD of three different donors per group. * p

Techniques Used: Cell Differentiation, Purification, Cell Culture, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Staining

40) Product Images from "TGF-β3-expressing CD4+CD25−LAG3+ regulatory T cells control humoral immune responses"

Article Title: TGF-β3-expressing CD4+CD25−LAG3+ regulatory T cells control humoral immune responses

Journal: Nature Communications

doi: 10.1038/ncomms7329

LAG3 + Treg suppress B-cell activation through TGF-β3. ( a ) Microarray comparisons of the gene expression profiles between B6 CD25 + Treg and B6 LAG3 + Treg. Normalized expression values from B6 CD4 + CD25 − CD45RB high naive T cells are depicted according to the colour scale shown. ( b ) Tgfb3 mRNA expression in sorted T-cell subsets taken from the spleens of B6 mice ( n =3 per group). ( c – e ) TGF-β1, 2 and 3 protein levels in the culture supernatants of the indicated T-cell subsets from B6 mice determined using ELISA. Cells were seeded at 1 × 10 5 cells per well ( n =4 per group). ( f ) CFSE-labelled B cells were stimulated with or without anti-IgM mAb in the presence or absence of rTGF-β3 ( n =3 per group). ( g ) Viability of anti-IgM-stimulated B cells in the presence or absence of rTGF-β3 (1 ng ml −1 ) was assessed by 7-AAD ( n =3 per group). ( h ) The effects of TGF-β3 on total IgG production in the culture supernatants of anti-CD40/IL-4-stimulated B cells, determined as in Fig. 1d ( n =3 per group). * P
Figure Legend Snippet: LAG3 + Treg suppress B-cell activation through TGF-β3. ( a ) Microarray comparisons of the gene expression profiles between B6 CD25 + Treg and B6 LAG3 + Treg. Normalized expression values from B6 CD4 + CD25 − CD45RB high naive T cells are depicted according to the colour scale shown. ( b ) Tgfb3 mRNA expression in sorted T-cell subsets taken from the spleens of B6 mice ( n =3 per group). ( c – e ) TGF-β1, 2 and 3 protein levels in the culture supernatants of the indicated T-cell subsets from B6 mice determined using ELISA. Cells were seeded at 1 × 10 5 cells per well ( n =4 per group). ( f ) CFSE-labelled B cells were stimulated with or without anti-IgM mAb in the presence or absence of rTGF-β3 ( n =3 per group). ( g ) Viability of anti-IgM-stimulated B cells in the presence or absence of rTGF-β3 (1 ng ml −1 ) was assessed by 7-AAD ( n =3 per group). ( h ) The effects of TGF-β3 on total IgG production in the culture supernatants of anti-CD40/IL-4-stimulated B cells, determined as in Fig. 1d ( n =3 per group). * P

Techniques Used: Activation Assay, Microarray, Expressing, Mouse Assay, Enzyme-linked Immunosorbent Assay

Related Articles

Enzyme-linked Immunosorbent Assay:

Article Title: Plasmacytoid dendritic cell deficiency in neonates enhances allergic airway inflammation via reduced production of IFN-α
Article Snippet: .. IL-4, IL-5, IL-10, IL-17A, IFN-γ, GM-CSF, and IgE ELISA kits were from BioLegend (San Diego, CA); IL-13 ELISA kit was from eBioscience (San Diego, CA); IL-33 and CCL20 ELISA kits were from R & D System (Minneapolis, MN); and IgG1 and IgG2a ELISA kits were from Bethyl (Montgomery, TX). .. Mouse antibodies used for flow cytometry were anti-siglec-F (clone E50-2440; BD, San Jose, CA), anti-Ly6G (clone 1A8; BD), anti-CD11c (clone HL3; BD), anti-CD3e (clone 145-2C11; BD), anti-CD19 (clone 1D3; BD), anti-CD11b (clone M1/70; BD), anti-CD64 (clone X54-5/7.1; BD), anti-CD45 (clone 30-F11; BD), anti-CD90.2 (clone 53-2.1; BD), anti-CD25 (clone PC61; BD), anti-ST2 (clone U29-93; BD), anti-B220 (clone RA3-6B2; BD), anti-siglec-H (clone eBio440c; eBioscience), anti-IA/IE (clone 2G9; BD), anti-PDCA1 (clone eBio927; eBioscience), anti-CD103 (clone M290; BD), anti-CD4 (clone H129.19; BD), anti-FoxP3 (clone MF23; BD), anti-GARP (clone F011-5; BioLegend), anti-IL-10 (clone JES5-16E3; BD), anti-TGF-β (clone TW7-20B9; BioLegend), anti-CD326 (clone G8.8; BD), and anti-CD31 (clone MEC 13.3; BD).

other:

Article Title: Further evidence associating IgG1, but not IgG2, with susceptibility to canine visceral leishmaniasis caused by Leishmania (L.) infantum chagasi-infection
Article Snippet: A number of studies in France, Spain, and Portugal, for example, reported the action of IgG2 with the development of symptomatic infections – therefore assuming that IgG2 plays a stronger immunopathogenic role than IgG1 [ , , , ].

Article Title: Further evidence associating IgG1, but not IgG2, with susceptibility to canine visceral leishmaniasis caused by Leishmania (L.) infantum chagasi-infection
Article Snippet: In addition, although the amplitude of the confidence interval of r coefficient ( ) indicates that this statistical correlation is not clinically significant, the determination coefficient (R2 = 0.3107) indicates that IgG and IgG1 within the DCG group are well correlated, e.g., vary together in a similar fashion, differently from IgG and IgG1 (R2 = 0.004475) and IgG and IgG2 (R2 = 0.04619) in the SCG group, and also differently from IgG and IgG2 values (R2 = 0.1204) in the DCG group.

Article Title: Further evidence associating IgG1, but not IgG2, with susceptibility to canine visceral leishmaniasis caused by Leishmania (L.) infantum chagasi-infection
Article Snippet: Deplazes et al. [ ], however, suggested the existence of a dichotomy in IgG1 and IgG2 responses in symptomatic and asymptomatic dogs – in the sense that IgG2 was associated with asymptomatic and IgG1 with symptomatic infections.

Article Title: The Impact of Aspergillus fumigatus Viability and Sensitization to Its Allergens on the Murine Allergic Asthma Phenotype
Article Snippet: For protocol (b), the serum was diluted as follows: IgA 1 : 1000, IgG1 1 : 5000, IgG2a 1 : 5000, and IgE 1 : 100, and the BALF was diluted as follows: IgA 1 : 10, IgG1 1 : 20, IgG2a 1 : 20, and IgE 1 : 2.

Infection:

Article Title: Further evidence associating IgG1, but not IgG2, with susceptibility to canine visceral leishmaniasis caused by Leishmania (L.) infantum chagasi-infection
Article Snippet: .. This also contrasted with reports from São Paulo and Minas Gerais States (southeastern Brazil) [ , ] that suggested that IgG2 is responsible for infection progression. .. A number of studies in France, Spain, and Portugal, for example, reported the action of IgG2 with the development of symptomatic infections – therefore assuming that IgG2 plays a stronger immunopathogenic role than IgG1 [ , , , ].

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 88
    Bethyl human igg elisa quantitation set
    On-bead magnetic screening. A one-bead one-compound (OBOC) library of thousands of unique peptoid compounds bound to TentaGel beads is incubated with control serum, here serum pooled from normal control subjects. The library is then incubated with anti-human <t>IgG-labeled</t> magnetic nanoparticles so that beads having bound IgG from the serum can be sorted out using a strong magnet. The library is initially depleted of beads that bind IgG from the control serum, and then incubated with target serum, here serum pooled from PD subjects. After incubation with the magnetic nanoparticles again, the newly magnetized beads, called ‘hits’, are isolated. Peptoid compounds are cleaved from each of the ‘hit’ beads and their sequences are assessed by MS/MS. These ‘hit’ compounds are then resynthesized and validated on <t>ELISA</t> plates for their ability to detect target IgG. (Reprinted by permission from Macmillan Publishers: from Zaman et al. 29 ). ELISA, enzyme-linked immunosorben assay.
    Human Igg Elisa Quantitation Set, supplied by Bethyl, used in various techniques. Bioz Stars score: 88/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human igg elisa quantitation set/product/Bethyl
    Average 88 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    human igg elisa quantitation set - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    Image Search Results


    On-bead magnetic screening. A one-bead one-compound (OBOC) library of thousands of unique peptoid compounds bound to TentaGel beads is incubated with control serum, here serum pooled from normal control subjects. The library is then incubated with anti-human IgG-labeled magnetic nanoparticles so that beads having bound IgG from the serum can be sorted out using a strong magnet. The library is initially depleted of beads that bind IgG from the control serum, and then incubated with target serum, here serum pooled from PD subjects. After incubation with the magnetic nanoparticles again, the newly magnetized beads, called ‘hits’, are isolated. Peptoid compounds are cleaved from each of the ‘hit’ beads and their sequences are assessed by MS/MS. These ‘hit’ compounds are then resynthesized and validated on ELISA plates for their ability to detect target IgG. (Reprinted by permission from Macmillan Publishers: from Zaman et al. 29 ). ELISA, enzyme-linked immunosorben assay.

    Journal: NPJ Parkinson's disease

    Article Title: Blood biomarker for Parkinson disease: peptoids

    doi: 10.1038/npjparkd.2016.12

    Figure Lengend Snippet: On-bead magnetic screening. A one-bead one-compound (OBOC) library of thousands of unique peptoid compounds bound to TentaGel beads is incubated with control serum, here serum pooled from normal control subjects. The library is then incubated with anti-human IgG-labeled magnetic nanoparticles so that beads having bound IgG from the serum can be sorted out using a strong magnet. The library is initially depleted of beads that bind IgG from the control serum, and then incubated with target serum, here serum pooled from PD subjects. After incubation with the magnetic nanoparticles again, the newly magnetized beads, called ‘hits’, are isolated. Peptoid compounds are cleaved from each of the ‘hit’ beads and their sequences are assessed by MS/MS. These ‘hit’ compounds are then resynthesized and validated on ELISA plates for their ability to detect target IgG. (Reprinted by permission from Macmillan Publishers: from Zaman et al. 29 ). ELISA, enzyme-linked immunosorben assay.

    Article Snippet: IgG levels of serum pools were measured using Human IgG ELISA Quantitation Set (Bethyl Laboratories, Montgomery, TX, USA).

    Techniques: Incubation, Labeling, Isolation, Mass Spectrometry, Enzyme-linked Immunosorbent Assay

    Chimeric antigen receptors (CAR) constructs for CD8+ T cells transduction ( A ) T cells were transduced with the lentiviruses to generate anti-CAIX CAR T cells, which are able to recognize CAIX positive RCC, and also secrete anti-PD-L1 IgG1 or IgG4 in the tumor microenvironment to block PD-1/PD-L1-induced T cell exhaustion. ( B ) Schematic representation of pHAGE lentiviral vectors encoding second-generation CARs fused with CD28 co-stimulatory endodomain. The anti-carbonic anhydrase IX (CAIX) or the Anti-B-cell maturation antigen (BCMA) scFv (as a negative control) were inserted after the eIFa promoter in order to express the CAR binding domain. The second cassette, after the Internal Ribosome Entry Site (IRES) sequence, encodes the secretable anti-PD-L1 IgG1 or IgG4 isotypes or the anti-severe acute respiratory syndrome (SARS) coronavirus IgG1 (negative control). LTR: long terminal repeat, eIFα: eukaryotic initiation factor alpha, scFv: single-chain variable fragment, C9 TAG: C9 peptide TETSQVAPA, IRES: Internal Ribosome Entry Site, WPRE: Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element. ( C ) Percentage of CAR T cells 14 days after transduction, representing the stable long-term expression of CAR by the integrated lentiviruses in CD8+ T cells. The CD8+ T cells were selected using Dynabeads CD8 Positive Isolation Kit (Life Technologies) and activated with Dynabeads Human T Cell Activator CD3/CD28 (Life Technologies) in the presence of IL-21 50 U/mL. IL-21 was added to the medium every 2 days. After 14 days, the CAR T cells were incubated with human CAIX-Fc or BCMA-Fc, followed by incubation with an APC conjugated anti-human Fc IgG (Southern Biotech) or goat-anti-mouse IgG Ab (Biolegend) and analyzed by flow cytometry. ( D ) Concentration of IgG secreted into the medium of transduced T cells evaluated by Human IgG ELISA Quantitation Set (Bethyl Laboratories). ( E ) Concentration of anti-PD-L1 antibodies in the supernatant of 293T Cells transduced with lentiviruses containing anti-CAIX or anti-BCMA CAR and anti-PD-L1 IgG1, anti-PD-L1 IgG4 or irrelevant anti-SARS IgG1 sequences. The antibodies in the supernatant were purified, biotinylated and incubated with 5 μg/mL of human PD-L1 pre-immobilized in the 96 wells MaxiSorp plate (Nunc). The biotinylated antibodies were detected by incubation with streptavidin-HRP and developed with TMB. The absorbance was read at λ = 450 nm. * P

    Journal: Oncotarget

    Article Title: Chimeric antigen receptor T cells secreting anti-PD-L1 antibodies more effectively regress renal cell carcinoma in a humanized mouse model

    doi: 10.18632/oncotarget.9114

    Figure Lengend Snippet: Chimeric antigen receptors (CAR) constructs for CD8+ T cells transduction ( A ) T cells were transduced with the lentiviruses to generate anti-CAIX CAR T cells, which are able to recognize CAIX positive RCC, and also secrete anti-PD-L1 IgG1 or IgG4 in the tumor microenvironment to block PD-1/PD-L1-induced T cell exhaustion. ( B ) Schematic representation of pHAGE lentiviral vectors encoding second-generation CARs fused with CD28 co-stimulatory endodomain. The anti-carbonic anhydrase IX (CAIX) or the Anti-B-cell maturation antigen (BCMA) scFv (as a negative control) were inserted after the eIFa promoter in order to express the CAR binding domain. The second cassette, after the Internal Ribosome Entry Site (IRES) sequence, encodes the secretable anti-PD-L1 IgG1 or IgG4 isotypes or the anti-severe acute respiratory syndrome (SARS) coronavirus IgG1 (negative control). LTR: long terminal repeat, eIFα: eukaryotic initiation factor alpha, scFv: single-chain variable fragment, C9 TAG: C9 peptide TETSQVAPA, IRES: Internal Ribosome Entry Site, WPRE: Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element. ( C ) Percentage of CAR T cells 14 days after transduction, representing the stable long-term expression of CAR by the integrated lentiviruses in CD8+ T cells. The CD8+ T cells were selected using Dynabeads CD8 Positive Isolation Kit (Life Technologies) and activated with Dynabeads Human T Cell Activator CD3/CD28 (Life Technologies) in the presence of IL-21 50 U/mL. IL-21 was added to the medium every 2 days. After 14 days, the CAR T cells were incubated with human CAIX-Fc or BCMA-Fc, followed by incubation with an APC conjugated anti-human Fc IgG (Southern Biotech) or goat-anti-mouse IgG Ab (Biolegend) and analyzed by flow cytometry. ( D ) Concentration of IgG secreted into the medium of transduced T cells evaluated by Human IgG ELISA Quantitation Set (Bethyl Laboratories). ( E ) Concentration of anti-PD-L1 antibodies in the supernatant of 293T Cells transduced with lentiviruses containing anti-CAIX or anti-BCMA CAR and anti-PD-L1 IgG1, anti-PD-L1 IgG4 or irrelevant anti-SARS IgG1 sequences. The antibodies in the supernatant were purified, biotinylated and incubated with 5 μg/mL of human PD-L1 pre-immobilized in the 96 wells MaxiSorp plate (Nunc). The biotinylated antibodies were detected by incubation with streptavidin-HRP and developed with TMB. The absorbance was read at λ = 450 nm. * P

    Article Snippet: Total level of IgG secreted into the medium of transduced cells was detected using Human IgG ELISA Quantitation Set (Bethyl Laboratories).

    Techniques: Construct, Transduction, Blocking Assay, Negative Control, Binding Assay, Sequencing, Expressing, Isolation, Incubation, Flow Cytometry, Cytometry, Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Purification