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Becton Dickinson anti mhc ii
Migrating CFSE + cDC transport viral RNA to the lung-draining LN and present antigens by <t>MHC-I</t> and MHC-II molecules. (A) MLN cDC were harvested 72 h post-RSV infection and sorted into four populations based on <t>CD103</t> expression and CFSE labeling.
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1) Product Images from "Respiratory Syncytial Virus-Induced Activation and Migration of Respiratory Dendritic Cells and Subsequent Antigen Presentation in the Lung-Draining Lymph Node ▿"

Article Title: Respiratory Syncytial Virus-Induced Activation and Migration of Respiratory Dendritic Cells and Subsequent Antigen Presentation in the Lung-Draining Lymph Node ▿

Journal: Journal of Virology

doi: 10.1128/JVI.00452-09

Migrating CFSE + cDC transport viral RNA to the lung-draining LN and present antigens by MHC-I and MHC-II molecules. (A) MLN cDC were harvested 72 h post-RSV infection and sorted into four populations based on CD103 expression and CFSE labeling.
Figure Legend Snippet: Migrating CFSE + cDC transport viral RNA to the lung-draining LN and present antigens by MHC-I and MHC-II molecules. (A) MLN cDC were harvested 72 h post-RSV infection and sorted into four populations based on CD103 expression and CFSE labeling.

Techniques Used: Infection, Expressing, Labeling

2) Product Images from "Chrysin Derivative CM1 and Exhibited Anti-Inflammatory Action by Upregulating Toll-Interacting Protein Expression in Lipopolysaccharide-Stimulated RAW264.7 Macrophage Cells"

Article Title: Chrysin Derivative CM1 and Exhibited Anti-Inflammatory Action by Upregulating Toll-Interacting Protein Expression in Lipopolysaccharide-Stimulated RAW264.7 Macrophage Cells

Journal: Molecules

doi: 10.3390/molecules26061532

CM1-mediated suppression of macrophage surface marker expression through upregulation of Tollip expression. Control and Tollip shRNA-transfected RAW264.7 cells were pretreated with CM1 (2 μg/mL) for 1 h and then exposed to LPS (200 ng/mL) for 24 h. Cells were stained with anti-CD80, anti-CD86, anti-MHC class I, and anti-MHC class II. The percentage of positive cells is shown in each panel. Data are presented as the mean ± SD for triplicate determinations of one representative plot among three independent experiments. Statistical analysis was performed by one-way ANOVA in conjunction with Tukey’s multiple test; n.s : no significance and *** p
Figure Legend Snippet: CM1-mediated suppression of macrophage surface marker expression through upregulation of Tollip expression. Control and Tollip shRNA-transfected RAW264.7 cells were pretreated with CM1 (2 μg/mL) for 1 h and then exposed to LPS (200 ng/mL) for 24 h. Cells were stained with anti-CD80, anti-CD86, anti-MHC class I, and anti-MHC class II. The percentage of positive cells is shown in each panel. Data are presented as the mean ± SD for triplicate determinations of one representative plot among three independent experiments. Statistical analysis was performed by one-way ANOVA in conjunction with Tukey’s multiple test; n.s : no significance and *** p

Techniques Used: Marker, Expressing, shRNA, Transfection, Staining

3) Product Images from "DC-Derived IL-10 Modulates Pro-inflammatory Cytokine Production and Promotes Induction of CD4+IL-10+ Regulatory T Cells during Plasmodium yoelii Infection"

Article Title: DC-Derived IL-10 Modulates Pro-inflammatory Cytokine Production and Promotes Induction of CD4+IL-10+ Regulatory T Cells during Plasmodium yoelii Infection

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2017.00152

Inactivation of IL-10 in IL-10 flox/flox /CD11c-cre mice results in enhanced IFN-γ and TNF-α production of CD11c + dendritic cells (DCs) during Plasmodium yoelii infection . (A) IL-10 secretion of LPS-stimulated CD11c + DCs isolated from spleen of IL-10 flox/flox (DC-IL-10 WT) and IL-10 flox/flox /CD11c-cre (DC-IL-10 KO) was analyzed by Luminex technology. IL-10 flox/flox and IL-10 flox/flox /CD11c-cre mice were infected with P. yoelii . At indicated time points postinfection, (B) the expression level (MFI) of MHC-II was analyzed on gated CD11c + CD11b + CD8 − DCs and CD11c + CD11b − CD8 + DCs, and the (C) frequencies of IFN-γ and (D) TNF-α-expressing CD11c + DCs were determined by flow cytometry. The gating strategy of CD11c + DCs and representative dot plots are shown in the upper panel. Results from at least two independent experiments with n = 6–16 mice per time point were summarized as mean ± SEM. Student’s t -test was used for statistical analysis (* p
Figure Legend Snippet: Inactivation of IL-10 in IL-10 flox/flox /CD11c-cre mice results in enhanced IFN-γ and TNF-α production of CD11c + dendritic cells (DCs) during Plasmodium yoelii infection . (A) IL-10 secretion of LPS-stimulated CD11c + DCs isolated from spleen of IL-10 flox/flox (DC-IL-10 WT) and IL-10 flox/flox /CD11c-cre (DC-IL-10 KO) was analyzed by Luminex technology. IL-10 flox/flox and IL-10 flox/flox /CD11c-cre mice were infected with P. yoelii . At indicated time points postinfection, (B) the expression level (MFI) of MHC-II was analyzed on gated CD11c + CD11b + CD8 − DCs and CD11c + CD11b − CD8 + DCs, and the (C) frequencies of IFN-γ and (D) TNF-α-expressing CD11c + DCs were determined by flow cytometry. The gating strategy of CD11c + DCs and representative dot plots are shown in the upper panel. Results from at least two independent experiments with n = 6–16 mice per time point were summarized as mean ± SEM. Student’s t -test was used for statistical analysis (* p

Techniques Used: Mouse Assay, Infection, Isolation, Luminex, Expressing, Flow Cytometry, Cytometry

4) Product Images from "Cutaneous immunosurveillance by self-renewing dermal ?? T cells"

Article Title: Cutaneous immunosurveillance by self-renewing dermal ?? T cells

Journal: The Journal of Experimental Medicine

doi: 10.1084/jem.20101824

Neutrophil recruitment to the site of BCG infection is compromised in the absence of IL-17-producing γδ T cells. (A, top left) Single-plane image from a dermal whole-mount stain depicting Gr1 + neutrophils at the site of BCG infection in ear skin of WT mice 24 h after infection. Bar, 500 µm. (top right) higher magnification of the boxed area in the top left panel depicting Gr1 + neutrophils around a deposit of BCG. Arrows point to BCG-containing Gr1 + neutrophils. Boxed inset shows a single Gr1 + neutrophil containing intracellular BCG bacilli. Bar, 50 µm. (bottom left) Extended focus image from a dermal whole-mount stain 36 h after BCG infection depicting BCG-incorporated Gr1 + neutrophils (arrows) in the lumen of a lymphatic vessel stained with anti–LYVE-1. Bars, 25 µm. (bottom right) Flow cytometry profiles of CD45 + leukocytes isolated from WT ear skin, 24h after i.d. infection with BCG-mCherry. CD45 + BCG-mCherry + cells were further evaluated for Ly6G and MHC II expression. Data are representative of two to four independent experiments. (B) Flow cytometry profiles of Gr1 + cells infiltrating the skin of WT mice (n = 4), 24h after i.d. injection with BCG or PBS. Cells were gated on CD45 + CD3 – . Data are representative of 3 independent experiments. (C) Absolute cell number of Gr1 + neutrophils isolated from ear skin of WT and TCRδ −/− mice (n = 14–17 per group), 24 h after i.d. infection with BCG. Data are representative of three independent experiments. (D) Copy number of 16S rRNA from BCG isolated from auricular draining LN of WT and TCRδ −/− mice (n = 5 per group), 3 d after i.d. infection. Data are representative of two independent experiments.
Figure Legend Snippet: Neutrophil recruitment to the site of BCG infection is compromised in the absence of IL-17-producing γδ T cells. (A, top left) Single-plane image from a dermal whole-mount stain depicting Gr1 + neutrophils at the site of BCG infection in ear skin of WT mice 24 h after infection. Bar, 500 µm. (top right) higher magnification of the boxed area in the top left panel depicting Gr1 + neutrophils around a deposit of BCG. Arrows point to BCG-containing Gr1 + neutrophils. Boxed inset shows a single Gr1 + neutrophil containing intracellular BCG bacilli. Bar, 50 µm. (bottom left) Extended focus image from a dermal whole-mount stain 36 h after BCG infection depicting BCG-incorporated Gr1 + neutrophils (arrows) in the lumen of a lymphatic vessel stained with anti–LYVE-1. Bars, 25 µm. (bottom right) Flow cytometry profiles of CD45 + leukocytes isolated from WT ear skin, 24h after i.d. infection with BCG-mCherry. CD45 + BCG-mCherry + cells were further evaluated for Ly6G and MHC II expression. Data are representative of two to four independent experiments. (B) Flow cytometry profiles of Gr1 + cells infiltrating the skin of WT mice (n = 4), 24h after i.d. injection with BCG or PBS. Cells were gated on CD45 + CD3 – . Data are representative of 3 independent experiments. (C) Absolute cell number of Gr1 + neutrophils isolated from ear skin of WT and TCRδ −/− mice (n = 14–17 per group), 24 h after i.d. infection with BCG. Data are representative of three independent experiments. (D) Copy number of 16S rRNA from BCG isolated from auricular draining LN of WT and TCRδ −/− mice (n = 5 per group), 3 d after i.d. infection. Data are representative of two independent experiments.

Techniques Used: Infection, Staining, Mouse Assay, Flow Cytometry, Cytometry, Isolation, Expressing, Injection

5) Product Images from "Critical Role of TLR4 in Human Metapneumovirus Mediated Innate Immune Responses and Disease Pathogenesis"

Article Title: Critical Role of TLR4 in Human Metapneumovirus Mediated Innate Immune Responses and Disease Pathogenesis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0078849

Lung dendritic cell characterization and lymphocyte proliferation in response to hMPV infection in TLR4 −/− mice. TLR4 −/− and WT mice were mock- or hMPV-infected, and sacrificed at week 1 and 2 p.i. to collect lungs. Single cell suspension was obtained and cDCs population enriched using CD11c-tagged magnetic beads isolation. CD40 and CD86 markers were analyzed in cells positive for CD11c and MHC-II by flow cytometry. Histograms showing the expression of costimulatory molecules (CD40, CD86 and MHCII) in cDC from WT mice (open histograms) and cDC from TLR4 −/− mice (dashed line open histograms), and isotype control (shaded histograms), are shown ( A ). Graph of baseline expression of costimulatory molecules in lung dendritic cells isolated from WT and TLR4 −/− mice ( B ). Dendritic cells (CD11c positive cells) were isolated from lungs of hMPV or mock-infected mice either WT or TLR4 −/− at day 7 p.i. and loaded with 10 µg/mL of OVA peptide for 2h prior to coculture with T cells. CD4 + T cells isolated from spleen of OT II mice were labeled with CFSE and cocultured with DCs at a ratio of 1∶2 (DC: T). T cell proliferation was measured by CFSE dilution (proliferating CD4 + cells have a lower CFSE intensity than non-proliferated control cells). Cultures without antigen served as controls. The bar graph shows the percentage of proliferating (CFSE low ) T cells among the total CD4 + T cell population. Data are expressed as mean ± SEM of four mice/group and represent one of two independent experiments ( C ).
Figure Legend Snippet: Lung dendritic cell characterization and lymphocyte proliferation in response to hMPV infection in TLR4 −/− mice. TLR4 −/− and WT mice were mock- or hMPV-infected, and sacrificed at week 1 and 2 p.i. to collect lungs. Single cell suspension was obtained and cDCs population enriched using CD11c-tagged magnetic beads isolation. CD40 and CD86 markers were analyzed in cells positive for CD11c and MHC-II by flow cytometry. Histograms showing the expression of costimulatory molecules (CD40, CD86 and MHCII) in cDC from WT mice (open histograms) and cDC from TLR4 −/− mice (dashed line open histograms), and isotype control (shaded histograms), are shown ( A ). Graph of baseline expression of costimulatory molecules in lung dendritic cells isolated from WT and TLR4 −/− mice ( B ). Dendritic cells (CD11c positive cells) were isolated from lungs of hMPV or mock-infected mice either WT or TLR4 −/− at day 7 p.i. and loaded with 10 µg/mL of OVA peptide for 2h prior to coculture with T cells. CD4 + T cells isolated from spleen of OT II mice were labeled with CFSE and cocultured with DCs at a ratio of 1∶2 (DC: T). T cell proliferation was measured by CFSE dilution (proliferating CD4 + cells have a lower CFSE intensity than non-proliferated control cells). Cultures without antigen served as controls. The bar graph shows the percentage of proliferating (CFSE low ) T cells among the total CD4 + T cell population. Data are expressed as mean ± SEM of four mice/group and represent one of two independent experiments ( C ).

Techniques Used: Infection, Mouse Assay, Magnetic Beads, Isolation, Flow Cytometry, Cytometry, Expressing, Labeling

6) Product Images from "Salivary Tick Cystatin OmC2 Targets Lysosomal Cathepsins S and C in Human Dendritic Cells"

Article Title: Salivary Tick Cystatin OmC2 Targets Lysosomal Cathepsins S and C in Human Dendritic Cells

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2017.00288

Effect of tick cystatin OmC2 on the expression of MHC II and CD86. The confocal images show MHC II (HLA-DR) in (A) non-differentiated MUTZ-3 cells, (B) differentiated MUTZ-3 cells, and (C) differentiated MUTZ-3 cells stimulated with LPS. The localization of endogenous cystatin C in the differentiated MUTZ-3 cells is shown in (B) . Bars: 10 μm. (D) LPS-induced maturation of differentiated MUTZ-3 cells in the presence of cystatin OmC2 is compared to non-treated cells (no added cystatin OmC2). (E) The histograms show HLA-DR and CD86 in the differentiated MUTZ-3 grown in the presence of cystatin OmC2 (no maturation with LPS). (D,E) The shadowed histograms represent the non-treated cells (no added cystatin OmC2 and no maturation with LPS) at the beginning of the experiment. A representative analysis of three independent biological replicates is shown. Mean fluorescence intensities (MFI) of labeled cell populations (geometric means) are shown in bar graphs.
Figure Legend Snippet: Effect of tick cystatin OmC2 on the expression of MHC II and CD86. The confocal images show MHC II (HLA-DR) in (A) non-differentiated MUTZ-3 cells, (B) differentiated MUTZ-3 cells, and (C) differentiated MUTZ-3 cells stimulated with LPS. The localization of endogenous cystatin C in the differentiated MUTZ-3 cells is shown in (B) . Bars: 10 μm. (D) LPS-induced maturation of differentiated MUTZ-3 cells in the presence of cystatin OmC2 is compared to non-treated cells (no added cystatin OmC2). (E) The histograms show HLA-DR and CD86 in the differentiated MUTZ-3 grown in the presence of cystatin OmC2 (no maturation with LPS). (D,E) The shadowed histograms represent the non-treated cells (no added cystatin OmC2 and no maturation with LPS) at the beginning of the experiment. A representative analysis of three independent biological replicates is shown. Mean fluorescence intensities (MFI) of labeled cell populations (geometric means) are shown in bar graphs.

Techniques Used: Expressing, Fluorescence, Labeling

7) Product Images from "RADIATION AND HOST RETINOIC ACID SIGNALING PROMOTE THE INDUCTION OF GUT-HOMING DONOR T CELLS AFTER ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION"

Article Title: RADIATION AND HOST RETINOIC ACID SIGNALING PROMOTE THE INDUCTION OF GUT-HOMING DONOR T CELLS AFTER ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

doi: 10.1111/ajt.15501

Radiation enhances RALDH enzyme activity and the induction of gut-homing donor T cells early after allogeneic BMT. (A-D) Balb/cJ mice received TBI at 900 cGy and were euthanized on day 2 after irradiation. 0.5×10 6 MLN or spleen cells were suspended in ALDEFLUOR assay buffer containing the activated ALDEFLUOR substrate and incubated for 45 minutes at 37 °C. Cells were then stained with anti-CD45, anti-CD11c and anti-MHC-II antibodies and 7-AAD. Live (7-AAD – ) CD45 + CD11c + MHC-II + ALDEFLUOR + cells were quantified by flow cytometry. Representative contour plots depicting ALDEFLUOR + DCs from spleen (A) and MLN (C) are shown. (B, D) Mean (± SEM) percentage of ALDEFLUOR + DCs from spleen (B) and MLN (D). Data are the cumulative results from 5 independent experiments; (E-H) Lethally irradiated (1300 cGy) or non-irradiated B6D2F1 mice were transplanted with 5 × 10 6 B6 BM together with B6 splenocytes (adjusted to yield a T-cell dose of 4 × 10 6 ). Mice in both groups were euthanized on day 4 after transplantation. The expression of integrin α4β7 on donor T cells isolated from MLNs and spleen of recipient mice was examined by gating on H-2 b+ H2 d– CD4 + or H-2 b+ H2 d– CD8 + donor T cells. (E, G) Representative contour plots for α4β7 expression on donor T cells isolated from MLNs (E) and spleen (G) are shown. (F, H) Mean (± SEM) percentage of α4β7 + donor T cells isolated from MLNs (F) and spleen (H) (n = 4 mice per group). Data are derived from one representative of 3 experiments. Statistics: **P ≤ .01, ***P ≤ .001, ****P ≤ .0001.
Figure Legend Snippet: Radiation enhances RALDH enzyme activity and the induction of gut-homing donor T cells early after allogeneic BMT. (A-D) Balb/cJ mice received TBI at 900 cGy and were euthanized on day 2 after irradiation. 0.5×10 6 MLN or spleen cells were suspended in ALDEFLUOR assay buffer containing the activated ALDEFLUOR substrate and incubated for 45 minutes at 37 °C. Cells were then stained with anti-CD45, anti-CD11c and anti-MHC-II antibodies and 7-AAD. Live (7-AAD – ) CD45 + CD11c + MHC-II + ALDEFLUOR + cells were quantified by flow cytometry. Representative contour plots depicting ALDEFLUOR + DCs from spleen (A) and MLN (C) are shown. (B, D) Mean (± SEM) percentage of ALDEFLUOR + DCs from spleen (B) and MLN (D). Data are the cumulative results from 5 independent experiments; (E-H) Lethally irradiated (1300 cGy) or non-irradiated B6D2F1 mice were transplanted with 5 × 10 6 B6 BM together with B6 splenocytes (adjusted to yield a T-cell dose of 4 × 10 6 ). Mice in both groups were euthanized on day 4 after transplantation. The expression of integrin α4β7 on donor T cells isolated from MLNs and spleen of recipient mice was examined by gating on H-2 b+ H2 d– CD4 + or H-2 b+ H2 d– CD8 + donor T cells. (E, G) Representative contour plots for α4β7 expression on donor T cells isolated from MLNs (E) and spleen (G) are shown. (F, H) Mean (± SEM) percentage of α4β7 + donor T cells isolated from MLNs (F) and spleen (H) (n = 4 mice per group). Data are derived from one representative of 3 experiments. Statistics: **P ≤ .01, ***P ≤ .001, ****P ≤ .0001.

Techniques Used: Activity Assay, Mouse Assay, Irradiation, Incubation, Staining, Flow Cytometry, Transplantation Assay, Expressing, Isolation, Derivative Assay

8) Product Images from "Survivin is released from cancer cells via exosomes"

Article Title: Survivin is released from cancer cells via exosomes

Journal: Apoptosis : an international journal on programmed cell death

doi: 10.1007/s10495-010-0534-4

Histogram profile of surface CD antigens on purified exosomes bound to anti-MHC class II ( a ) or CD9 ( b ) -coupled aldehyde beads. Presence of LAMP1, Hsp70, CD9, CD54 and Survivin antigens were analyzed using exosomes bound to anti-MHC class II- or CD9-coupled
Figure Legend Snippet: Histogram profile of surface CD antigens on purified exosomes bound to anti-MHC class II ( a ) or CD9 ( b ) -coupled aldehyde beads. Presence of LAMP1, Hsp70, CD9, CD54 and Survivin antigens were analyzed using exosomes bound to anti-MHC class II- or CD9-coupled

Techniques Used: Purification

9) Product Images from "Enhancement of immunostimulatory properties of exosomal vaccines by incorporation of fusion-competent G protein of vesicular stomatitis virus"

Article Title: Enhancement of immunostimulatory properties of exosomal vaccines by incorporation of fusion-competent G protein of vesicular stomatitis virus

Journal: Vaccine

doi: 10.1016/j.vaccine.2008.04.069

Activation of splenic DC by ELVs. (A) Freshly isolated splenic DC were incubated with LPS (1 μg/ml) or with 10 μg/ml of ELVs (ELV-G, ELV-G mut or O-ELV) for 24 h. Cells were collected and stained for MHC II, CD11c, CD40, CD80, and CD86. CD11c + MHCII + cells were analyzed for CD40, CD80, and CD86 expression levels by flow cytometry. Numbers indicated in the upper right of the histograms represent the Geometric log MFI. One representative experiment of three is shown. (B) Cell-free supernatants were analyzed in triplicate for the presence of IL-12p70 (pg/ml) by ELISA. Data present the mean from three independent experiments ± standard deviation. * p
Figure Legend Snippet: Activation of splenic DC by ELVs. (A) Freshly isolated splenic DC were incubated with LPS (1 μg/ml) or with 10 μg/ml of ELVs (ELV-G, ELV-G mut or O-ELV) for 24 h. Cells were collected and stained for MHC II, CD11c, CD40, CD80, and CD86. CD11c + MHCII + cells were analyzed for CD40, CD80, and CD86 expression levels by flow cytometry. Numbers indicated in the upper right of the histograms represent the Geometric log MFI. One representative experiment of three is shown. (B) Cell-free supernatants were analyzed in triplicate for the presence of IL-12p70 (pg/ml) by ELISA. Data present the mean from three independent experiments ± standard deviation. * p

Techniques Used: Activation Assay, Isolation, Incubation, Staining, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Standard Deviation

10) Product Images from "Mononuclear Phagocyte Activation Is Associated With the Immunopathology of Psoriasis"

Article Title: Mononuclear Phagocyte Activation Is Associated With the Immunopathology of Psoriasis

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2020.00478

Frequency of monocyte subsets in patients with psoriasis vulgaris and healthy individuals. Skin biopsies (4 mm) and PBMCs were obtained, and ex-vivo staining for MHC II, CD14, and CD16 was performed. (A) Representative plots of monocyte subsets in PBMCs of a patient with psoriasis. (B) Frequency of circulating monocyte subsets in healthy subjects (HS) ( n = 6) and patients with psoriasis ( n = 25). (C) Representative plots of monocyte subsets in a lesion biopsy from a patient with psoriasis. (D) Mean fluorescent intensity (MFI) of CD14 in skin biopsies from HS ( n = 8) and in lesions from patients with psoriasis ( n = 23). Statistical comparisons were performed using the Kruskal-Wallis test and Dunn's post-test * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.
Figure Legend Snippet: Frequency of monocyte subsets in patients with psoriasis vulgaris and healthy individuals. Skin biopsies (4 mm) and PBMCs were obtained, and ex-vivo staining for MHC II, CD14, and CD16 was performed. (A) Representative plots of monocyte subsets in PBMCs of a patient with psoriasis. (B) Frequency of circulating monocyte subsets in healthy subjects (HS) ( n = 6) and patients with psoriasis ( n = 25). (C) Representative plots of monocyte subsets in a lesion biopsy from a patient with psoriasis. (D) Mean fluorescent intensity (MFI) of CD14 in skin biopsies from HS ( n = 8) and in lesions from patients with psoriasis ( n = 23). Statistical comparisons were performed using the Kruskal-Wallis test and Dunn's post-test * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Techniques Used: Ex Vivo, Staining

11) Product Images from "Intracellular Delivery of Lipopolysaccharide Induces Effective Th1-Immune Responses Independent of IL-12"

Article Title: Intracellular Delivery of Lipopolysaccharide Induces Effective Th1-Immune Responses Independent of IL-12

Journal: PLoS ONE

doi: 10.1371/journal.pone.0068671

LPS-liposomes induce co-stimulatory molecules in DCs as efficiently as LPS without inducing excessive production of TNF-α. (A) WT mice were intravenously treated with LPS (10 µg per mouse) or LPS-liposomes (10 µg per mouse). At 6 h after treatment, splenocytes were harvested and the expression of MHC-II, CD40, CD80, and CD86 on CD11c + splenic DCs was analyzed by flow cytometry. Splenic DCs from PBS treated mice were overlaid as control (open histograms). Percentage (%) are average of three independent experiments. n = 3 animals per group. The values represent means ± S.E.M * P
Figure Legend Snippet: LPS-liposomes induce co-stimulatory molecules in DCs as efficiently as LPS without inducing excessive production of TNF-α. (A) WT mice were intravenously treated with LPS (10 µg per mouse) or LPS-liposomes (10 µg per mouse). At 6 h after treatment, splenocytes were harvested and the expression of MHC-II, CD40, CD80, and CD86 on CD11c + splenic DCs was analyzed by flow cytometry. Splenic DCs from PBS treated mice were overlaid as control (open histograms). Percentage (%) are average of three independent experiments. n = 3 animals per group. The values represent means ± S.E.M * P

Techniques Used: Mouse Assay, Expressing, Flow Cytometry, Cytometry

12) Product Images from "Epstein-Barr Virus Promotes Interferon-? Production by Plasmacytoid Dendritic Cells"

Article Title: Epstein-Barr Virus Promotes Interferon-? Production by Plasmacytoid Dendritic Cells

Journal: Arthritis and rheumatism

doi: 10.1002/art.27408

IFN-α production by pDC requires MHC class II engagement
Figure Legend Snippet: IFN-α production by pDC requires MHC class II engagement

Techniques Used:

13) Product Images from "Low doses of cholera toxin and its mediator cAMP induce CTLA-2 secretion by dendritic cells to enhance regulatory T cell conversion"

Article Title: Low doses of cholera toxin and its mediator cAMP induce CTLA-2 secretion by dendritic cells to enhance regulatory T cell conversion

Journal: PLoS ONE

doi: 10.1371/journal.pone.0178114

CT and cAMP induces both CTLA-2α and -2β during DC maturation. BM-DCs were differentially matured with the indicated stimuli for 16h unless otherwise specified. (A) Principal component analysis comparing untreated BM-DCs, DCs matured for 6h with the Th2-inducing stimuli TNF, the Trypanosoma brucei antigens Mitat or mVSG, and the Th1-inducing LPS from our previous study [ 17 ] with CT hi (1 μg/ml) matured Th17-inducing DCs investigated in this study. (B) Heat map displaying the top 25 genes regulated in DCs after CT hi or TNF stimulation compared with untreated DCs. Data show single time point values of a single microarray. (C) Densitometric analysis of CTLA-2α and -2β expression levels determined by RT-sqPCR normalized to β-actin and relative to untreated control for n = 4 experiments; CT lo (0.1 μg/ml). (D) CT hi stimulation of BM-DCs ± the cAMP inhibitor KH7 before RT-sqPCR normalized to β-actin of n = 3 independent and pooled experiments. (E) DC maturation analysis by flow cytometry of CD86 and MHC-II on CD11c positive cells. (F) Statistical evaluation of D with respect to untreated control for n = 5 experiments. (G) IL-1β, IL-6 and IL-23 secretion by differentially stimulated DCs measured by ELISA n = 3 experiments. Statistical analysis was performed with respect to untreated control. Error bars represent mean ± SD. One Way ANOVA, Dunnett post-test. *p
Figure Legend Snippet: CT and cAMP induces both CTLA-2α and -2β during DC maturation. BM-DCs were differentially matured with the indicated stimuli for 16h unless otherwise specified. (A) Principal component analysis comparing untreated BM-DCs, DCs matured for 6h with the Th2-inducing stimuli TNF, the Trypanosoma brucei antigens Mitat or mVSG, and the Th1-inducing LPS from our previous study [ 17 ] with CT hi (1 μg/ml) matured Th17-inducing DCs investigated in this study. (B) Heat map displaying the top 25 genes regulated in DCs after CT hi or TNF stimulation compared with untreated DCs. Data show single time point values of a single microarray. (C) Densitometric analysis of CTLA-2α and -2β expression levels determined by RT-sqPCR normalized to β-actin and relative to untreated control for n = 4 experiments; CT lo (0.1 μg/ml). (D) CT hi stimulation of BM-DCs ± the cAMP inhibitor KH7 before RT-sqPCR normalized to β-actin of n = 3 independent and pooled experiments. (E) DC maturation analysis by flow cytometry of CD86 and MHC-II on CD11c positive cells. (F) Statistical evaluation of D with respect to untreated control for n = 5 experiments. (G) IL-1β, IL-6 and IL-23 secretion by differentially stimulated DCs measured by ELISA n = 3 experiments. Statistical analysis was performed with respect to untreated control. Error bars represent mean ± SD. One Way ANOVA, Dunnett post-test. *p

Techniques Used: Microarray, Expressing, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

Model on the differential effects of CT and cAMP on DC-directed induction of Th17 or Foxp3 + Treg cells. Immature DCs treated with high (CT hi ) or low (CT lo ) doses of cholera toxin or its known secondary mediator cAMP induced partial DC maturation measured as upregulation of MHC II- and costimulatory molecules such as CD86. CT lo -DCs and cAMP-DCs produced no pro-inflammatory cytokines (semi-mature DCs) but instead the tolerogenic molecule CTLA-2. Under Treg polarizing conditions in vitro , using naive TCR-transgenic CD4 + T cells, CTLA-2 produced by CT lo -DCs and cAMP-DCs supports the TGF-β mediated conversion into Foxp3 + Tregs. In contrast, CT hi -DCs also released CTLA-2 but additionally the Th17-polarizing cytokines IL-1β, IL-6 and IL-23 (mature DCs). Consequently CT hi -DCs mainly generated Th17 cells in vitro . In vivo , a polyclonal repertoire of T cells triggered by CT lo -DCs and cAMP-DCs results in both Th17 and Tregs activation pointing to competing effects at early or late time points in the EAE model.
Figure Legend Snippet: Model on the differential effects of CT and cAMP on DC-directed induction of Th17 or Foxp3 + Treg cells. Immature DCs treated with high (CT hi ) or low (CT lo ) doses of cholera toxin or its known secondary mediator cAMP induced partial DC maturation measured as upregulation of MHC II- and costimulatory molecules such as CD86. CT lo -DCs and cAMP-DCs produced no pro-inflammatory cytokines (semi-mature DCs) but instead the tolerogenic molecule CTLA-2. Under Treg polarizing conditions in vitro , using naive TCR-transgenic CD4 + T cells, CTLA-2 produced by CT lo -DCs and cAMP-DCs supports the TGF-β mediated conversion into Foxp3 + Tregs. In contrast, CT hi -DCs also released CTLA-2 but additionally the Th17-polarizing cytokines IL-1β, IL-6 and IL-23 (mature DCs). Consequently CT hi -DCs mainly generated Th17 cells in vitro . In vivo , a polyclonal repertoire of T cells triggered by CT lo -DCs and cAMP-DCs results in both Th17 and Tregs activation pointing to competing effects at early or late time points in the EAE model.

Techniques Used: Produced, In Vitro, Transgenic Assay, Generated, In Vivo, Activation Assay

14) Product Images from "Serine leucocyte proteinase inhibitor-treated monocyte inhibits human CD4+ lymphocyte proliferation"

Article Title: Serine leucocyte proteinase inhibitor-treated monocyte inhibits human CD4+ lymphocyte proliferation

Journal: Immunology

doi: 10.1111/j.1365-2567.2011.03451.x

Serine leucocyte proteinase inhibitor (SLPI) modifies the phenotype and cytokine secretion of monocyte. (a) Expression of CD86, CD54, CD18, MHC class I and class II in monocytes treated with SLPI (4 μg/ml) for 18 hr. Data are expressed in the
Figure Legend Snippet: Serine leucocyte proteinase inhibitor (SLPI) modifies the phenotype and cytokine secretion of monocyte. (a) Expression of CD86, CD54, CD18, MHC class I and class II in monocytes treated with SLPI (4 μg/ml) for 18 hr. Data are expressed in the

Techniques Used: Expressing

15) Product Images from "IL-12/23p40 overproduction by dendritic cells leads to an increased Th1 and Th17 polarization in a model of Yersinia enterocolitica-induced reactive arthritis in TNFRp55-/- mice"

Article Title: IL-12/23p40 overproduction by dendritic cells leads to an increased Th1 and Th17 polarization in a model of Yersinia enterocolitica-induced reactive arthritis in TNFRp55-/- mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0193573

IL-12/23p40 + dendritic cells in spleen of WT and TNFRp55 -/- mice on ReA onset. Splenic cells were obtained from WT and TNFRp55 -/- mice on day 14 after Ye infection (ReA onset). Dendritic cells (DCs) were stained for cell surface CD11c and MHC-II, and analyzed by flow cytometry. (A) Absolute DC number in the spleen of Ye-infected WT and TNFRp55 -/- mice are presented. Splenic DCs of uninfected (PBS) mice were used as controls. (B) Representative dot plot showing analysis of IL-12/23p40 + DCs (CD11c + MHC-II + gate) in splenocytes from control (PBS), and Ye-infected WT and TNFRp55 -/- mice. The cells were stimulated with PMA/Ionomycin and brefeldin for 5 h, and then stained for cell surface CD11c and MHC-II, and intracellular IL-12/23p40, and then analyzed by flow cytometry. The numbers in the plots indicate the percentages of labeled cells in representative mice. (C) Percentage of IL-12/23p40 + DCs of the sum of three independent experiments. Each symbol represents an individual mouse; horizontal lines indicate the mean ± SEM. *** P
Figure Legend Snippet: IL-12/23p40 + dendritic cells in spleen of WT and TNFRp55 -/- mice on ReA onset. Splenic cells were obtained from WT and TNFRp55 -/- mice on day 14 after Ye infection (ReA onset). Dendritic cells (DCs) were stained for cell surface CD11c and MHC-II, and analyzed by flow cytometry. (A) Absolute DC number in the spleen of Ye-infected WT and TNFRp55 -/- mice are presented. Splenic DCs of uninfected (PBS) mice were used as controls. (B) Representative dot plot showing analysis of IL-12/23p40 + DCs (CD11c + MHC-II + gate) in splenocytes from control (PBS), and Ye-infected WT and TNFRp55 -/- mice. The cells were stimulated with PMA/Ionomycin and brefeldin for 5 h, and then stained for cell surface CD11c and MHC-II, and intracellular IL-12/23p40, and then analyzed by flow cytometry. The numbers in the plots indicate the percentages of labeled cells in representative mice. (C) Percentage of IL-12/23p40 + DCs of the sum of three independent experiments. Each symbol represents an individual mouse; horizontal lines indicate the mean ± SEM. *** P

Techniques Used: Mouse Assay, Infection, Staining, Flow Cytometry, Cytometry, Labeling

16) Product Images from "Bacillus subtilis Spores as Vaccine Adjuvants: Further Insights into the Mechanisms of Action"

Article Title: Bacillus subtilis Spores as Vaccine Adjuvants: Further Insights into the Mechanisms of Action

Journal: PLoS ONE

doi: 10.1371/journal.pone.0087454

Maturation of dendritic cells after stimulation with B. subtilis spores. (A) Induction of pro-inflammatory cytokines after stimulation with spores. BMDCs from C57BL/6 mice were treated with PBS (control group), LT1 and live or heat-killed spores at a MOI of 10.000. Supernatants were collected after 24 h, and the IL-1β, IL12p70 and TNF-α concentrations were quantified by ELISA. (B) Expression of surface molecules by CD11c + cells following exposure to spores. BMDCs were treated as described above and the surface expression levels of CD40, MHC-I and MHC-II were measured by flow cytometry. Gray-filled histograms represent samples incubated with PBS and open histograms represent samples incubated with spores. The MFI for both conditions is shown in the upper left in each histogram. (C) Graphical comparison of the expression of CD40, MHC-I and MHC-II. Data represent mean values ± s.e.m. of two independent experiments. * p
Figure Legend Snippet: Maturation of dendritic cells after stimulation with B. subtilis spores. (A) Induction of pro-inflammatory cytokines after stimulation with spores. BMDCs from C57BL/6 mice were treated with PBS (control group), LT1 and live or heat-killed spores at a MOI of 10.000. Supernatants were collected after 24 h, and the IL-1β, IL12p70 and TNF-α concentrations were quantified by ELISA. (B) Expression of surface molecules by CD11c + cells following exposure to spores. BMDCs were treated as described above and the surface expression levels of CD40, MHC-I and MHC-II were measured by flow cytometry. Gray-filled histograms represent samples incubated with PBS and open histograms represent samples incubated with spores. The MFI for both conditions is shown in the upper left in each histogram. (C) Graphical comparison of the expression of CD40, MHC-I and MHC-II. Data represent mean values ± s.e.m. of two independent experiments. * p

Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Cytometry, Incubation

Intracellular cell staining and flow cytometry analysis of p24-specific CD4 + and CD8 + T cells. BALB/c mice were immunized (three doses on days 0, 14 and 28) with p24 protein (700 ng), p24 (10 µg) admixed with LT1 (1 µg), p24 (700 ng) admixed with spores (2×10 9 ) or p24 (700 ng) admixed with heat-killed spores (2×10 9 ). Spleen cells were collected two weeks after the last dose, and the detection of IL-4 producing p24-specific CD4 + T cells (A) and IFN-γ producing p24-specific CD4 + (B) and CD8 + T cells (C) was carried out after stimulation with MHC class I or II restricted p24 peptide and cell surface staining for CD8 (FITC), CD4 (Cy) and intracellular staining for IL-4 or IFN-γ (PE). The percentage of each population is indicated in the upper right corners and the frequencies in all groups in the absence of stimulus were below 0.70% (data not shown). Measurements were performed in duplicate for each individual sample, and the data are representative of two independent experiments.
Figure Legend Snippet: Intracellular cell staining and flow cytometry analysis of p24-specific CD4 + and CD8 + T cells. BALB/c mice were immunized (three doses on days 0, 14 and 28) with p24 protein (700 ng), p24 (10 µg) admixed with LT1 (1 µg), p24 (700 ng) admixed with spores (2×10 9 ) or p24 (700 ng) admixed with heat-killed spores (2×10 9 ). Spleen cells were collected two weeks after the last dose, and the detection of IL-4 producing p24-specific CD4 + T cells (A) and IFN-γ producing p24-specific CD4 + (B) and CD8 + T cells (C) was carried out after stimulation with MHC class I or II restricted p24 peptide and cell surface staining for CD8 (FITC), CD4 (Cy) and intracellular staining for IL-4 or IFN-γ (PE). The percentage of each population is indicated in the upper right corners and the frequencies in all groups in the absence of stimulus were below 0.70% (data not shown). Measurements were performed in duplicate for each individual sample, and the data are representative of two independent experiments.

Techniques Used: Staining, Flow Cytometry, Cytometry, Mouse Assay

17) Product Images from "Chrysin Derivative CM1 and Exhibited Anti-Inflammatory Action by Upregulating Toll-Interacting Protein Expression in Lipopolysaccharide-Stimulated RAW264.7 Macrophage Cells"

Article Title: Chrysin Derivative CM1 and Exhibited Anti-Inflammatory Action by Upregulating Toll-Interacting Protein Expression in Lipopolysaccharide-Stimulated RAW264.7 Macrophage Cells

Journal: Molecules

doi: 10.3390/molecules26061532

CM1-mediated suppression of macrophage surface marker expression through upregulation of Tollip expression. Control and Tollip shRNA-transfected RAW264.7 cells were pretreated with CM1 (2 μg/mL) for 1 h and then exposed to LPS (200 ng/mL) for 24 h. Cells were stained with anti-CD80, anti-CD86, anti-MHC class I, and anti-MHC class II. The percentage of positive cells is shown in each panel. Data are presented as the mean ± SD for triplicate determinations of one representative plot among three independent experiments. Statistical analysis was performed by one-way ANOVA in conjunction with Tukey’s multiple test; n.s : no significance and *** p
Figure Legend Snippet: CM1-mediated suppression of macrophage surface marker expression through upregulation of Tollip expression. Control and Tollip shRNA-transfected RAW264.7 cells were pretreated with CM1 (2 μg/mL) for 1 h and then exposed to LPS (200 ng/mL) for 24 h. Cells were stained with anti-CD80, anti-CD86, anti-MHC class I, and anti-MHC class II. The percentage of positive cells is shown in each panel. Data are presented as the mean ± SD for triplicate determinations of one representative plot among three independent experiments. Statistical analysis was performed by one-way ANOVA in conjunction with Tukey’s multiple test; n.s : no significance and *** p

Techniques Used: Marker, Expressing, shRNA, Transfection, Staining

18) Product Images from "Lineage targeted MHC-II transgenic mice demonstrate the role of dendritic cells in bacterial driven colitis"

Article Title: Lineage targeted MHC-II transgenic mice demonstrate the role of dendritic cells in bacterial driven colitis

Journal: Inflammatory bowel diseases

doi: 10.1002/ibd.23000

Immunohistochemistry for CD11c and MHC-II in Ntg and Tg mice CD11c and MHC-II expressing cells in the colon of Ntg (A, B) and Tg (C, D) mice. CD11c + cells in lamina propria (A, C) and MHC-II + cells only noted in Tg animals (D) compared to no MHC-II staining detected in Ntg animals (B).
Figure Legend Snippet: Immunohistochemistry for CD11c and MHC-II in Ntg and Tg mice CD11c and MHC-II expressing cells in the colon of Ntg (A, B) and Tg (C, D) mice. CD11c + cells in lamina propria (A, C) and MHC-II + cells only noted in Tg animals (D) compared to no MHC-II staining detected in Ntg animals (B).

Techniques Used: Immunohistochemistry, Mouse Assay, Expressing, Staining

Transgene constructs and expression analysis in transgenic animal tissues (A) Construct design for CD11cTg mice. I-A b β was placed under the control of the CD11c promotor with rabbit β-Globin Gene used as a marker for transgene expression (B) Two constructs, one each for I-A b α and I-A b β, were co-injected into MHC-II-deficient (I-E/I-Aβ-deficient). Both MHC-II chains were under control of promotor elements of fatty acid binding protein to direct expression to small intestine (FABPL) (C) Representative dot plots showing staining with anti- MHC-II antibody versus anti-CD11c antibody in splenocytes (top row) and MLN cell suspensions (bottom row) from C57BL/6 (WT), Rag2 −/− , CD11cTg/Rag2 −/− and CD11cNtg/Rag2 −/− mice (indicated in figure). Dot plots are gated through a FSC/SSC gate that excluded small (presumably dead) cells. (D) Histograms show MHC-II expression of CD11c + cells in spleen (left) and MLN (right). (E) RT-PCR showing transgene expression of the I-A b α construct, the I-A b β construct and HPRT in various tissues of EpithTg line 6258.
Figure Legend Snippet: Transgene constructs and expression analysis in transgenic animal tissues (A) Construct design for CD11cTg mice. I-A b β was placed under the control of the CD11c promotor with rabbit β-Globin Gene used as a marker for transgene expression (B) Two constructs, one each for I-A b α and I-A b β, were co-injected into MHC-II-deficient (I-E/I-Aβ-deficient). Both MHC-II chains were under control of promotor elements of fatty acid binding protein to direct expression to small intestine (FABPL) (C) Representative dot plots showing staining with anti- MHC-II antibody versus anti-CD11c antibody in splenocytes (top row) and MLN cell suspensions (bottom row) from C57BL/6 (WT), Rag2 −/− , CD11cTg/Rag2 −/− and CD11cNtg/Rag2 −/− mice (indicated in figure). Dot plots are gated through a FSC/SSC gate that excluded small (presumably dead) cells. (D) Histograms show MHC-II expression of CD11c + cells in spleen (left) and MLN (right). (E) RT-PCR showing transgene expression of the I-A b α construct, the I-A b β construct and HPRT in various tissues of EpithTg line 6258.

Techniques Used: Construct, Expressing, Transgenic Assay, Mouse Assay, Marker, Injection, Binding Assay, Staining, Reverse Transcription Polymerase Chain Reaction

19) Product Images from "IL-12/23p40 overproduction by dendritic cells leads to an increased Th1 and Th17 polarization in a model of Yersinia enterocolitica-induced reactive arthritis in TNFRp55-/- mice"

Article Title: IL-12/23p40 overproduction by dendritic cells leads to an increased Th1 and Th17 polarization in a model of Yersinia enterocolitica-induced reactive arthritis in TNFRp55-/- mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0193573

IL-12/23p40 + dendritic cells in spleen of WT and TNFRp55 -/- mice on ReA onset. Splenic cells were obtained from WT and TNFRp55 -/- mice on day 14 after Ye infection (ReA onset). Dendritic cells (DCs) were stained for cell surface CD11c and MHC-II, and analyzed by flow cytometry. (A) Absolute DC number in the spleen of Ye-infected WT and TNFRp55 -/- mice are presented. Splenic DCs of uninfected (PBS) mice were used as controls. (B) Representative dot plot showing analysis of IL-12/23p40 + DCs (CD11c + MHC-II + gate) in splenocytes from control (PBS), and Ye-infected WT and TNFRp55 -/- mice. The cells were stimulated with PMA/Ionomycin and brefeldin for 5 h, and then stained for cell surface CD11c and MHC-II, and intracellular IL-12/23p40, and then analyzed by flow cytometry. The numbers in the plots indicate the percentages of labeled cells in representative mice. (C) Percentage of IL-12/23p40 + DCs of the sum of three independent experiments. Each symbol represents an individual mouse; horizontal lines indicate the mean ± SEM. *** P
Figure Legend Snippet: IL-12/23p40 + dendritic cells in spleen of WT and TNFRp55 -/- mice on ReA onset. Splenic cells were obtained from WT and TNFRp55 -/- mice on day 14 after Ye infection (ReA onset). Dendritic cells (DCs) were stained for cell surface CD11c and MHC-II, and analyzed by flow cytometry. (A) Absolute DC number in the spleen of Ye-infected WT and TNFRp55 -/- mice are presented. Splenic DCs of uninfected (PBS) mice were used as controls. (B) Representative dot plot showing analysis of IL-12/23p40 + DCs (CD11c + MHC-II + gate) in splenocytes from control (PBS), and Ye-infected WT and TNFRp55 -/- mice. The cells were stimulated with PMA/Ionomycin and brefeldin for 5 h, and then stained for cell surface CD11c and MHC-II, and intracellular IL-12/23p40, and then analyzed by flow cytometry. The numbers in the plots indicate the percentages of labeled cells in representative mice. (C) Percentage of IL-12/23p40 + DCs of the sum of three independent experiments. Each symbol represents an individual mouse; horizontal lines indicate the mean ± SEM. *** P

Techniques Used: Mouse Assay, Infection, Staining, Flow Cytometry, Cytometry, Labeling

20) Product Images from "Impact of Mammalian Target of Rapamycin Inhibition on Lymphoid Homing and Tolerogenic Function of Nanoparticle-Labeled Dendritic Cells following Allogeneic Hematopoietic Cell Transplantation 1"

Article Title: Impact of Mammalian Target of Rapamycin Inhibition on Lymphoid Homing and Tolerogenic Function of Nanoparticle-Labeled Dendritic Cells following Allogeneic Hematopoietic Cell Transplantation 1

Journal: Journal of immunology (Baltimore, Md. : 1950)

doi:

Migratory and stimulatory capacity of SPIO-labeled DC remains intact. A , CD40, CD80, CD86, and MHC class II expression on DC with or without SPIO labeling as indicated for the respective line. Experiments were repeated three times. B , Migratory capacity
Figure Legend Snippet: Migratory and stimulatory capacity of SPIO-labeled DC remains intact. A , CD40, CD80, CD86, and MHC class II expression on DC with or without SPIO labeling as indicated for the respective line. Experiments were repeated three times. B , Migratory capacity

Techniques Used: Labeling, Expressing

21) Product Images from "Foxp3-Expressing CD103+ Regulatory T Cells Accumulate in Dendritic Cell Aggregates of the Colonic Mucosa in Murine Transfer Colitis"

Article Title: Foxp3-Expressing CD103+ Regulatory T Cells Accumulate in Dendritic Cell Aggregates of the Colonic Mucosa in Murine Transfer Colitis

Journal: The American Journal of Pathology

doi: 10.2353/ajpath.2006.050228

Intraepithelial DCs are associated with DC aggregates and display a mature phenotype. A and B: Double staining of CD11c (red) to label DCs and laminin (green) to visualize the basal membrane. Intraepithelial CD11c + DCs (red) located at the luminal side of the basal membrane are predominantly found in association with DC aggregates ( arrowheads in B ). Note a DC passing a gap in the basal membrane ( arrow in B ). Single immunohistochemical staining of MHC-II ( C ) reveals predominant expression of MHC-II ( C ), CD40 ( D ), CD80 ( E ), and CD86 ( F ) in DC aggregates. Original magnifications: ×10 ( A , C–F ), ×40 ( B ).
Figure Legend Snippet: Intraepithelial DCs are associated with DC aggregates and display a mature phenotype. A and B: Double staining of CD11c (red) to label DCs and laminin (green) to visualize the basal membrane. Intraepithelial CD11c + DCs (red) located at the luminal side of the basal membrane are predominantly found in association with DC aggregates ( arrowheads in B ). Note a DC passing a gap in the basal membrane ( arrow in B ). Single immunohistochemical staining of MHC-II ( C ) reveals predominant expression of MHC-II ( C ), CD40 ( D ), CD80 ( E ), and CD86 ( F ) in DC aggregates. Original magnifications: ×10 ( A , C–F ), ×40 ( B ).

Techniques Used: Double Staining, Immunohistochemistry, Staining, Expressing

22) Product Images from "DNA Encoding an HIV-1 Gag/Human Lysosome-Associated Membrane Protein-1 Chimera Elicits a Broad Cellular and Humoral Immune Response in Rhesus Macaques"

Article Title: DNA Encoding an HIV-1 Gag/Human Lysosome-Associated Membrane Protein-1 Chimera Elicits a Broad Cellular and Humoral Immune Response in Rhesus Macaques

Journal: PLoS ONE

doi: 10.1371/journal.pone.0000135

Expression and trafficking of mouse and human LAMP/ gag. (A) Western blot analysis of human 293 cells transfected with mLAMP/ gag and hLAMP/ gag plasmids. Samples were probed with an anti-Gag monoclonal antibody. The molecular weight markers are indicated on the right. (B) DCEK cells were transfected with the hLAMP/ gag plasmid and stained with anti-Gag (red) or anti-MHC II (green) monoclonal antibodies. Digitally merged image shows co-localization of the hLAMP/ gag chimera- and MHC II-containing compartments (yellow).
Figure Legend Snippet: Expression and trafficking of mouse and human LAMP/ gag. (A) Western blot analysis of human 293 cells transfected with mLAMP/ gag and hLAMP/ gag plasmids. Samples were probed with an anti-Gag monoclonal antibody. The molecular weight markers are indicated on the right. (B) DCEK cells were transfected with the hLAMP/ gag plasmid and stained with anti-Gag (red) or anti-MHC II (green) monoclonal antibodies. Digitally merged image shows co-localization of the hLAMP/ gag chimera- and MHC II-containing compartments (yellow).

Techniques Used: Expressing, Western Blot, Transfection, Molecular Weight, Plasmid Preparation, Staining

23) Product Images from "MFG-E8 Regulates the Immunogenic Potential of Dendritic Cells Primed with Necrotic Cell-Mediated Inflammatory Signals"

Article Title: MFG-E8 Regulates the Immunogenic Potential of Dendritic Cells Primed with Necrotic Cell-Mediated Inflammatory Signals

Journal: PLoS ONE

doi: 10.1371/journal.pone.0039607

MFG-E8-deficiecy confers BMDC with an activated phenotype under steady and inflammatory conditions. (A) Five different sequences of small interfering RNA for MFG-E8 (MFG-E8i #1-#5) or a control gene (control siRNA) were introduced into immature BMDC for 48 h and the mRNA levels of MFG-E8 were quantified by RT-PCR. The mature DC (mDC) stimulated with CD40 ligand (CD40L) serve as a control due to their low MFG-E8 expression. (B) The expression levels of CD86 on CD11c + DC were evaluated by flow cytometry. The percentage (left) and mean fluorescence intensity (MFI) of CD86 among CD11c+DC was shown as a representative data and statistical analysis (n = 3), respectively. (C) The expression levels of CD83 and MHC-II were evaluated in DC from wild-type (WT) and MFG-E8-decifient (MFG-E8-KO) mice. The representative data (upper) and statistical analysis of three independent experiments (bottom) are shown. * p
Figure Legend Snippet: MFG-E8-deficiecy confers BMDC with an activated phenotype under steady and inflammatory conditions. (A) Five different sequences of small interfering RNA for MFG-E8 (MFG-E8i #1-#5) or a control gene (control siRNA) were introduced into immature BMDC for 48 h and the mRNA levels of MFG-E8 were quantified by RT-PCR. The mature DC (mDC) stimulated with CD40 ligand (CD40L) serve as a control due to their low MFG-E8 expression. (B) The expression levels of CD86 on CD11c + DC were evaluated by flow cytometry. The percentage (left) and mean fluorescence intensity (MFI) of CD86 among CD11c+DC was shown as a representative data and statistical analysis (n = 3), respectively. (C) The expression levels of CD83 and MHC-II were evaluated in DC from wild-type (WT) and MFG-E8-decifient (MFG-E8-KO) mice. The representative data (upper) and statistical analysis of three independent experiments (bottom) are shown. * p

Techniques Used: Small Interfering RNA, Reverse Transcription Polymerase Chain Reaction, Expressing, Flow Cytometry, Cytometry, Fluorescence, Mouse Assay

24) Product Images from "Neonatal immune responses to TLR2 stimulation: Influence of maternal atopy on Foxp3 and IL-10 expression"

Article Title: Neonatal immune responses to TLR2 stimulation: Influence of maternal atopy on Foxp3 and IL-10 expression

Journal: Respiratory Research

doi: 10.1186/1465-9921-7-40

A+B. Lymphocyte proliferation following addition of anti-MHC II or anti-CD4 ab is unchanged in unstimulated CBMC and following stimulation with the innate stimulus Ppg. Following addition of anti-MHC II or anti-CD4 ab, lymphocyte proliferation is decreased after stimulation with the allergen Derf1 (p
Figure Legend Snippet: A+B. Lymphocyte proliferation following addition of anti-MHC II or anti-CD4 ab is unchanged in unstimulated CBMC and following stimulation with the innate stimulus Ppg. Following addition of anti-MHC II or anti-CD4 ab, lymphocyte proliferation is decreased after stimulation with the allergen Derf1 (p

Techniques Used:

Related Articles

other:

Article Title: Immature Dendritic Cells Phagocytose Apoptotic Cells via ?v?5 and CD36, and Cross-present Antigens to Cytotoxic T Lymphocytes
Article Snippet: Antibodies to the following proteins were used: CD8-PE, CD14-PE, HLA-DR–PE, HLA-DR–biotin ( Becton Dickinson ), IgG2b (clone 6603001; Coulter Corp., Hialeah, FL), CD8 (CRL 8014; American Type Culture Collection, Rockville, MD), CD83 (clone HB15a; Coulter Corp.), MHC I (W6/ 32, ATCC clone HB95), CD36 (clone FA6; obtained from the fifth international workshop on leukocyte differentiation antigens), αv (clone CLB-706, Chemicon International, Inc., Temecula, CA; clone 69.6.5, Coulter Corp.), β1 (clone 6S6; Chemicon International, Inc.), β3 (clone SZ21, Coulter Corp.; clone RUU-PL 7F12, Becton Dickinson ), β5 (clone B5-IVF2; Upstate Biotechnology, Inc., Lake Placid, NY), αv β3 (clone 23C6; PharMingen , San Diego, CA), αv β5 (clone P1F6; Chemicon International, Inc.), CD71 (Dako Corp., Carpinteria, CA), mannose receptor (clone 3.2PB1; a gift from A. Lanzavecchia, Basel Institute, Sweden), and influenza nucleoprotein (ATCC clone HB85).

Article Title: MHC class II stabilization at the surface of human dendritic cells is the result of maturation-dependent MARCH I down-regulation
Article Snippet: Anti-mature HLA-DR (L243 clone; BD Pharmingen) ( ); rabbit polyclonal anti-HLA-DR (DRAB; P. Cresswell, New Haven, CT); anti-HLA-DR (clone ISCR3); anti-HLA-DR β-chain specific (clone XD5); rabbit polyclonal anti-HLA-DRα; anti-CD74 (clone PIN-1 and clone BU45); anti-MARCH I and anti-MARCH VIII, both a generous gift from Laurent Coscoy (University of California, Berkeley, CA) and Klaus Früh (Oregon Health and Science University, Portland, OR); anti-ubiquitin (clone P4D1; Covance); anti-GFP (clone JL-8; Clontech); rabbit polyclonal anti-HLA-DM (Lars Karlson, San Diego, CA); anti-EEA1 (clone 14), anti-human LAMP-1 (clone H4A3) from BD Transduction Laboratory; anti-TfR (clone OKT9; M. Vidal, Montpellier, France); anti-CD1a (clone BL6) from Beckman Coulter; allophycocyanin-conjugated L243 and biotin-conjugated anti-CD86 (clone IT2.2) from BD Pharmingen.

Expressing:

Article Title: Visualization of Myelin Basic Protein (Mbp) T Cell Epitopes in Multiple Sclerosis Lesions Using a Monoclonal Antibody Specific for the Human Histocompatibility Leukocyte Antigen (Hla)-Dr2-Mbp 85-99 Complex
Article Snippet: .. To quantitate DR expression, cells were incubated directly with a biotin-labeled anti–HLA-DR mAb (mAb L243; Becton Dickinson), followed by incubation with R-PE–labeled streptavidin as described above. ..

Incubation:

Article Title: Visualization of Myelin Basic Protein (Mbp) T Cell Epitopes in Multiple Sclerosis Lesions Using a Monoclonal Antibody Specific for the Human Histocompatibility Leukocyte Antigen (Hla)-Dr2-Mbp 85-99 Complex
Article Snippet: .. To quantitate DR expression, cells were incubated directly with a biotin-labeled anti–HLA-DR mAb (mAb L243; Becton Dickinson), followed by incubation with R-PE–labeled streptavidin as described above. ..

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  • 86
    Becton Dickinson anti mhc class ii
    Turnover of SI-LP and MLN <t>CD103</t> + and CD103 − DC. (A) Mice were injected i.p. with 2 mg BrdU, and the percentage of BrdU + CD103 + and CD103 − DCs <t>(MHC</t> class II + CD11c + ) in the SI-LP and MLN was determined by flow cytometry at the times indicated. Results are the mean and SD of three to seven independent experiments with two mice in each time point except the 72-h time point, which was performed once. (B) BrdU and Ki67 staining on SI-LP and MLN CD103 + and CD103 − DCs (MHC class II + CD11c + ) was assessed by flow cytometry 3 and 24 h after BrdU injection. Plots are from one representative experiment of three performed. (C) SI from CD45.2 + mice (graft) was transplanted into CD45.1 + recipients (host) as previously described ( 32 ). At days 6 and 45, host and graft intestine were sectioned and stained with antibodies to CD45.2 (red), CD11c (blue), and MHC class II (green). Immunohistochemistry of the 45-d graft is shown. Arrows point to host-derived CD45.2 − MHC class II hi CD11c + DCs. Bar, 25 μm. The graph is percentage of host-derived (CD45.2 negative) CD11c + MHC class II hi cells in graft and host small intestinal villus. Results are mean and SD ( n = 3 mice per group).
    Anti Mhc Class Ii, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Migrating CFSE + cDC transport viral RNA to the lung-draining LN and present antigens by <t>MHC-I</t> and MHC-II molecules. (A) MLN cDC were harvested 72 h post-RSV infection and sorted into four populations based on <t>CD103</t> expression and CFSE labeling.
    Anti Mhc Ii, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Peripheral activated macrophages, especially those of the classically activated M1 phenotype, are enriched in MPTP-induced PD mice. (A) Representative flow cytometry analysis of peripheral macrophage subpopulations in saline and MPTP mice. Macrophage subpopulations were characterized on the basis of <t>MHC</t> II (PE) and <t>CD11b</t> (FITC) staining. (B) Total number of macrophages in spleen as a percentage of total cells at different times. (C) The ratio of CD11b+ MHC II hi (M1) macrophages to CD11b+ MHC II low (M2) macrophages at different times. Values are presented as the mean±SEM, * p
    Pe Rat Anti Mouse Mhc Ii, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    <t>MHC</t> Class I restriction assay. MHC class I restriction assay was performed by a cytotoxic assay of testing SV 95 -specific CTL lines against SV 95–1 peptide-pulsed T2 cells with the addition of blocking antibody against MHC class I molecule (anti-HLA-A2, <t>BB7.2),</t> or MHC class II molecule (anti-MHC class II, IVA 12). SV 95–6 specific CTL clone was a representative among the SV 95 CTL lines. The cytotoxicity of SV 95–6 specific CTLs was completely blocked by MHC I antibody, but not at all by MHC II antibody.
    Anti Mhc Ii Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Turnover of SI-LP and MLN CD103 + and CD103 − DC. (A) Mice were injected i.p. with 2 mg BrdU, and the percentage of BrdU + CD103 + and CD103 − DCs (MHC class II + CD11c + ) in the SI-LP and MLN was determined by flow cytometry at the times indicated. Results are the mean and SD of three to seven independent experiments with two mice in each time point except the 72-h time point, which was performed once. (B) BrdU and Ki67 staining on SI-LP and MLN CD103 + and CD103 − DCs (MHC class II + CD11c + ) was assessed by flow cytometry 3 and 24 h after BrdU injection. Plots are from one representative experiment of three performed. (C) SI from CD45.2 + mice (graft) was transplanted into CD45.1 + recipients (host) as previously described ( 32 ). At days 6 and 45, host and graft intestine were sectioned and stained with antibodies to CD45.2 (red), CD11c (blue), and MHC class II (green). Immunohistochemistry of the 45-d graft is shown. Arrows point to host-derived CD45.2 − MHC class II hi CD11c + DCs. Bar, 25 μm. The graph is percentage of host-derived (CD45.2 negative) CD11c + MHC class II hi cells in graft and host small intestinal villus. Results are mean and SD ( n = 3 mice per group).

    Journal: The Journal of Experimental Medicine

    Article Title: Small intestinal CD103+ dendritic cells display unique functional properties that are conserved between mice and humans

    doi: 10.1084/jem.20080414

    Figure Lengend Snippet: Turnover of SI-LP and MLN CD103 + and CD103 − DC. (A) Mice were injected i.p. with 2 mg BrdU, and the percentage of BrdU + CD103 + and CD103 − DCs (MHC class II + CD11c + ) in the SI-LP and MLN was determined by flow cytometry at the times indicated. Results are the mean and SD of three to seven independent experiments with two mice in each time point except the 72-h time point, which was performed once. (B) BrdU and Ki67 staining on SI-LP and MLN CD103 + and CD103 − DCs (MHC class II + CD11c + ) was assessed by flow cytometry 3 and 24 h after BrdU injection. Plots are from one representative experiment of three performed. (C) SI from CD45.2 + mice (graft) was transplanted into CD45.1 + recipients (host) as previously described ( 32 ). At days 6 and 45, host and graft intestine were sectioned and stained with antibodies to CD45.2 (red), CD11c (blue), and MHC class II (green). Immunohistochemistry of the 45-d graft is shown. Arrows point to host-derived CD45.2 − MHC class II hi CD11c + DCs. Bar, 25 μm. The graph is percentage of host-derived (CD45.2 negative) CD11c + MHC class II hi cells in graft and host small intestinal villus. Results are mean and SD ( n = 3 mice per group).

    Article Snippet: Cell staining reagents The following reagents were used to stain murine cells: unconjugated or directly conjugated anti-CD103 (M290), anti-α4β7 (DATK32), anti-CD62L (MEL-14), anti-MHC class II (2G9), anti-CD11c (N418), and rat IgG2a and IgG2b isotype controls (BD Biosciences); perCP-Cy5.5, Alexa 700–conjugated anti-CD11c (N418), and Pacific blue–conjugated anti–MHC Class II (M5/114.15.2; BioLegend); anti-CCR7 (4B12; eBiosciences); recombinant mouse E-selectin human Fc chimera (R & D Systems); anti-FcRII/III (2.4G2), anti-CD40 (FGK45), anti-CD86 (GL1), and anti-CCR9 antibody (7E7, ( ); purified from hybridomas); Cy5-labeled donkey anti–human IgG F(ab′)2 (Jackson ImmunoResearch Laboratories); biotinylated mouse anti–rat IgG2a (RG7/1.30), biotinylated mouse anti–rat IgG2b (G15-337), PE-labeled Ki67, streptavidin-conjugated PE, PECy7, PerCP Cy5.5, and the BrdU FITC flow kit (BD Biosciences); streptavidin–alexa 488 (Invitrogen); and DAPI (Sigma-Aldrich).

    Techniques: Mouse Assay, Injection, Flow Cytometry, Cytometry, Staining, Immunohistochemistry, Derivative Assay

    Localization of CD103 + and CD103 − MHC class II + CD11c + cells within the small intestinal mucosa. (A and B) Expression of CD103 by DCs in small intestinal villus and SILT. (A) Images show overlays of CD103 (red) in combination with CD11c (blue) and MHC class II (green), with insets showing DAPI staining. Arrows point toward MHC class II hi CD11c + DCs coexpressing CD103. Bars, 50 μm. (B) The graph is percentage of MHC class II hi CD11c + cells in SI villus or dome region of SILT that express CD103. Results are mean and SD ( n = 5 mice per group). ***, P

    Journal: The Journal of Experimental Medicine

    Article Title: Small intestinal CD103+ dendritic cells display unique functional properties that are conserved between mice and humans

    doi: 10.1084/jem.20080414

    Figure Lengend Snippet: Localization of CD103 + and CD103 − MHC class II + CD11c + cells within the small intestinal mucosa. (A and B) Expression of CD103 by DCs in small intestinal villus and SILT. (A) Images show overlays of CD103 (red) in combination with CD11c (blue) and MHC class II (green), with insets showing DAPI staining. Arrows point toward MHC class II hi CD11c + DCs coexpressing CD103. Bars, 50 μm. (B) The graph is percentage of MHC class II hi CD11c + cells in SI villus or dome region of SILT that express CD103. Results are mean and SD ( n = 5 mice per group). ***, P

    Article Snippet: Cell staining reagents The following reagents were used to stain murine cells: unconjugated or directly conjugated anti-CD103 (M290), anti-α4β7 (DATK32), anti-CD62L (MEL-14), anti-MHC class II (2G9), anti-CD11c (N418), and rat IgG2a and IgG2b isotype controls (BD Biosciences); perCP-Cy5.5, Alexa 700–conjugated anti-CD11c (N418), and Pacific blue–conjugated anti–MHC Class II (M5/114.15.2; BioLegend); anti-CCR7 (4B12; eBiosciences); recombinant mouse E-selectin human Fc chimera (R & D Systems); anti-FcRII/III (2.4G2), anti-CD40 (FGK45), anti-CD86 (GL1), and anti-CCR9 antibody (7E7, ( ); purified from hybridomas); Cy5-labeled donkey anti–human IgG F(ab′)2 (Jackson ImmunoResearch Laboratories); biotinylated mouse anti–rat IgG2a (RG7/1.30), biotinylated mouse anti–rat IgG2b (G15-337), PE-labeled Ki67, streptavidin-conjugated PE, PECy7, PerCP Cy5.5, and the BrdU FITC flow kit (BD Biosciences); streptavidin–alexa 488 (Invitrogen); and DAPI (Sigma-Aldrich).

    Techniques: Expressing, Staining, Mouse Assay

    Migrating CFSE + cDC transport viral RNA to the lung-draining LN and present antigens by MHC-I and MHC-II molecules. (A) MLN cDC were harvested 72 h post-RSV infection and sorted into four populations based on CD103 expression and CFSE labeling.

    Journal: Journal of Virology

    Article Title: Respiratory Syncytial Virus-Induced Activation and Migration of Respiratory Dendritic Cells and Subsequent Antigen Presentation in the Lung-Draining Lymph Node ▿

    doi: 10.1128/JVI.00452-09

    Figure Lengend Snippet: Migrating CFSE + cDC transport viral RNA to the lung-draining LN and present antigens by MHC-I and MHC-II molecules. (A) MLN cDC were harvested 72 h post-RSV infection and sorted into four populations based on CD103 expression and CFSE labeling.

    Article Snippet: The cells were stained in PBS containing 2% fetal calf serum, 2 mM EDTA, and 0.02% NaN3 with the following monoclonal antibodies: anti-CD4 (L3T4), anti-CD8α (clone 53-6.7), anti-CD11b (clone M1/70), anti-CD11c (clone HL3), anti-CD19 (clone 1D3), anti-CD40 (clone 3/23), anti-CD45R (B220, clone RA3-6B2), anti-CD80 (clone 16-10A1), anti-CD86 (clone GL1), anti-CD103 (clone M290), and anti-MHC-II (I-Ab /I-Eb ) (clone M5/114.15.2), obtained from BD Biosciences (San Diego, CA), and anti-mPDCA-1 (clone JF05-1C2.4.1), obtained from Miltenyi Biotec (Germany).

    Techniques: Infection, Expressing, Labeling

    Peripheral activated macrophages, especially those of the classically activated M1 phenotype, are enriched in MPTP-induced PD mice. (A) Representative flow cytometry analysis of peripheral macrophage subpopulations in saline and MPTP mice. Macrophage subpopulations were characterized on the basis of MHC II (PE) and CD11b (FITC) staining. (B) Total number of macrophages in spleen as a percentage of total cells at different times. (C) The ratio of CD11b+ MHC II hi (M1) macrophages to CD11b+ MHC II low (M2) macrophages at different times. Values are presented as the mean±SEM, * p

    Journal: Frontiers in Aging Neuroscience

    Article Title: Partial Depletion of Peripheral M1 Macrophages Reverses Motor Deficits in MPTP-Treated Mouse by Suppressing Neuroinflammation and Dopaminergic Neurodegeneration

    doi: 10.3389/fnagi.2018.00160

    Figure Lengend Snippet: Peripheral activated macrophages, especially those of the classically activated M1 phenotype, are enriched in MPTP-induced PD mice. (A) Representative flow cytometry analysis of peripheral macrophage subpopulations in saline and MPTP mice. Macrophage subpopulations were characterized on the basis of MHC II (PE) and CD11b (FITC) staining. (B) Total number of macrophages in spleen as a percentage of total cells at different times. (C) The ratio of CD11b+ MHC II hi (M1) macrophages to CD11b+ MHC II low (M2) macrophages at different times. Values are presented as the mean±SEM, * p

    Article Snippet: After being washed in PBS, the remaining cells were resuspended in 300 μl of PBS and stained for 30 min at 4°C with different combinations of FITC rat anti-mouse CD11b (cat.no.557396, 1:200, BD Pharmingen; Groves et al., ) and PE rat anti-mouse MHC II (cat.no.557000, 1:200, BD Pharmingen; Wang et al., ).

    Techniques: Mouse Assay, Flow Cytometry, Cytometry, Staining

    Clodronate liposome treatments successfully depleted peripheral macrophages in the spleen. (A) Flow cytometry analysis shows the percentage of MHC II+ CD11b+macrophages in the spleens of the PBS lipo-saline, clodronate lipo-saline, PBS lipo-MPTP, and clodronate lipo-MPTP groups at 7 days after MPTP injection. (B) Total splenic macrophage numbers as a percentage of total leukocytes in different experimental groups. (C) The ratio of CD11b+ MHC II hi (M1) macrophages to CD11b+ MHC II low (M2) macrophages in different experimental groups. (D) Analysis of iNOS and Arg-1 protein levels in the spleen at 7 days after MPTP injection. (E) Quantification of the densitometric value of the iNOS protein bands is shown, normalized to β-actin. (F) Quantification of the densitometric value of the Arg-1 protein bands, normalized to β-actin, is also shown. The gels were run under the same experimental conditions. Values are mean±SEM. * p

    Journal: Frontiers in Aging Neuroscience

    Article Title: Partial Depletion of Peripheral M1 Macrophages Reverses Motor Deficits in MPTP-Treated Mouse by Suppressing Neuroinflammation and Dopaminergic Neurodegeneration

    doi: 10.3389/fnagi.2018.00160

    Figure Lengend Snippet: Clodronate liposome treatments successfully depleted peripheral macrophages in the spleen. (A) Flow cytometry analysis shows the percentage of MHC II+ CD11b+macrophages in the spleens of the PBS lipo-saline, clodronate lipo-saline, PBS lipo-MPTP, and clodronate lipo-MPTP groups at 7 days after MPTP injection. (B) Total splenic macrophage numbers as a percentage of total leukocytes in different experimental groups. (C) The ratio of CD11b+ MHC II hi (M1) macrophages to CD11b+ MHC II low (M2) macrophages in different experimental groups. (D) Analysis of iNOS and Arg-1 protein levels in the spleen at 7 days after MPTP injection. (E) Quantification of the densitometric value of the iNOS protein bands is shown, normalized to β-actin. (F) Quantification of the densitometric value of the Arg-1 protein bands, normalized to β-actin, is also shown. The gels were run under the same experimental conditions. Values are mean±SEM. * p

    Article Snippet: After being washed in PBS, the remaining cells were resuspended in 300 μl of PBS and stained for 30 min at 4°C with different combinations of FITC rat anti-mouse CD11b (cat.no.557396, 1:200, BD Pharmingen; Groves et al., ) and PE rat anti-mouse MHC II (cat.no.557000, 1:200, BD Pharmingen; Wang et al., ).

    Techniques: Flow Cytometry, Cytometry, Injection

    MHC Class I restriction assay. MHC class I restriction assay was performed by a cytotoxic assay of testing SV 95 -specific CTL lines against SV 95–1 peptide-pulsed T2 cells with the addition of blocking antibody against MHC class I molecule (anti-HLA-A2, BB7.2), or MHC class II molecule (anti-MHC class II, IVA 12). SV 95–6 specific CTL clone was a representative among the SV 95 CTL lines. The cytotoxicity of SV 95–6 specific CTLs was completely blocked by MHC I antibody, but not at all by MHC II antibody.

    Journal: Frontiers in Molecular Biosciences

    Article Title: Novel Survivin Peptides Screened With Computer Algorithm Induce Cytotoxic T Lymphocytes With Higher Cytotoxic Efficiency to Cancer Cells

    doi: 10.3389/fmolb.2020.570003

    Figure Lengend Snippet: MHC Class I restriction assay. MHC class I restriction assay was performed by a cytotoxic assay of testing SV 95 -specific CTL lines against SV 95–1 peptide-pulsed T2 cells with the addition of blocking antibody against MHC class I molecule (anti-HLA-A2, BB7.2), or MHC class II molecule (anti-MHC class II, IVA 12). SV 95–6 specific CTL clone was a representative among the SV 95 CTL lines. The cytotoxicity of SV 95–6 specific CTLs was completely blocked by MHC I antibody, but not at all by MHC II antibody.

    Article Snippet: MHC Restriction AssayMajor histocompatibility complex restriction assay was performed using the same method of above cytotoxicity assay except an addition of blocking antibodies including anti-MHC class I antibody (clone BB7.2, BD Pharmingen, San Diego, CA, United States), or anti-MHC II antibody (clone G46-6, Pharmingen, San Diego, CA, United States).

    Techniques: Restriction Assay, Blocking Assay