Structured Review

Becton Dickinson hla dr
Differentiation and survival molecule expression by lamina propria (LP) and bone marrow (BM) plasma cells (PC). (A) Representative example (top series of histograms) of LPPC and BMPC (CD38 h cells) expression of several differentiation molecules (CD19, <t>CD20,</t> <t>HLA-DR,</t> and VS38c) and survival associated molecules (CD95, Bcl-2). Negative control histograms are shown in grey. (B) Mean (SEM) of several experiments (n≥5) for each marker, expressed as a percentage of positive PC. *p
Hla Dr, 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
https://www.bioz.com/result/hla dr/product/Becton Dickinson
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
hla dr - by Bioz Stars, 2021-09
86/100 stars

Images

1) Product Images from "Isolation, maturational level, and functional capacity of human colon lamina propria plasma cells"

Article Title: Isolation, maturational level, and functional capacity of human colon lamina propria plasma cells

Journal: Gut

doi:

Differentiation and survival molecule expression by lamina propria (LP) and bone marrow (BM) plasma cells (PC). (A) Representative example (top series of histograms) of LPPC and BMPC (CD38 h cells) expression of several differentiation molecules (CD19, CD20, HLA-DR, and VS38c) and survival associated molecules (CD95, Bcl-2). Negative control histograms are shown in grey. (B) Mean (SEM) of several experiments (n≥5) for each marker, expressed as a percentage of positive PC. *p
Figure Legend Snippet: Differentiation and survival molecule expression by lamina propria (LP) and bone marrow (BM) plasma cells (PC). (A) Representative example (top series of histograms) of LPPC and BMPC (CD38 h cells) expression of several differentiation molecules (CD19, CD20, HLA-DR, and VS38c) and survival associated molecules (CD95, Bcl-2). Negative control histograms are shown in grey. (B) Mean (SEM) of several experiments (n≥5) for each marker, expressed as a percentage of positive PC. *p

Techniques Used: Expressing, Negative Control, Marker

2) Product Images from "In vivo pre-activation of monocytes in patients with axial spondyloarthritis"

Article Title: In vivo pre-activation of monocytes in patients with axial spondyloarthritis

Journal: Arthritis Research & Therapy

doi: 10.1186/s13075-015-0694-2

Elevated percentage of classic monocytes in patients with axial spondyloarthritis (axSpA). a The percentage of classic CD14 ++ CD16 − (M1), intermediate CD14 ++ CD16 + (M2), and non-classic CD14 + CD16 + (M3) monocytes was determined by fluorescence-activated cell sorting in whole blood of controls, patients with axSpA, and patients with rheumatoid arthritis (RA). The percentage of classic, intermediate, and non-classic monocytes was determined among monocytes which were gated according to forward (FSC) and side (SSC) scatter (R1) and HLA-DR expression (R2). b The percentage of classic, intermediate, and non-classic monocytes (individual measurements and median) is shown from measurements in 12 controls (C), 20 patients with axSpA (SpA), and 12 patients with RA (RA). * P
Figure Legend Snippet: Elevated percentage of classic monocytes in patients with axial spondyloarthritis (axSpA). a The percentage of classic CD14 ++ CD16 − (M1), intermediate CD14 ++ CD16 + (M2), and non-classic CD14 + CD16 + (M3) monocytes was determined by fluorescence-activated cell sorting in whole blood of controls, patients with axSpA, and patients with rheumatoid arthritis (RA). The percentage of classic, intermediate, and non-classic monocytes was determined among monocytes which were gated according to forward (FSC) and side (SSC) scatter (R1) and HLA-DR expression (R2). b The percentage of classic, intermediate, and non-classic monocytes (individual measurements and median) is shown from measurements in 12 controls (C), 20 patients with axSpA (SpA), and 12 patients with RA (RA). * P

Techniques Used: Fluorescence, FACS, Expressing

3) Product Images from "A general Fc engineering platform for the next generation of antibody therapeutics"

Article Title: A general Fc engineering platform for the next generation of antibody therapeutics

Journal: Theranostics

doi: 10.7150/thno.51299

Engineered CD40 agonist antibodies displayed divergent FcγRIIa and FcγRIIb-dependent agonistic activity. (A) Different concentrations of CD40 agonist antibody variants were added to CD40 reporter cells in the presence of human FcγRIIb, FcγRIIa R131 or FcγRIIa H131 -expressing HEK293T cells, the activation of CD40 after 24 h stimulation was indicated by the expression of GFP of the reporter cell and analyzed by flow cytometry. (B) B cells were stimulated by different concentrations of NK003 variants and the upregulation of CD23, CD86 or HLA-DR was analyzed by flow cytometry. MFI: Mean Fluorescence Intensity. (C) Schematic of OVA-specific CD8 + T cell response model. FcγR/CD40-humanized mice were adoptively transferred with OT-I cells, and then immunized with DEC-OVA in the presence of CD40 antibody variants, expansion of OT-I cells was analyzed by flow cytometry. (D) Quantification of OT-I cells (left) and the percentage of OT-I cells among CD8 + T cells (right) as in OVA-specific CD8 + T cell response model. Each circle represents an individual mouse. Bars represent the mean ± SEM. ** p ≤0.01, *** p ≤0.001, **** p ≤0.0001; One-way ANOVA Tukey's multiple comparisons test was used.
Figure Legend Snippet: Engineered CD40 agonist antibodies displayed divergent FcγRIIa and FcγRIIb-dependent agonistic activity. (A) Different concentrations of CD40 agonist antibody variants were added to CD40 reporter cells in the presence of human FcγRIIb, FcγRIIa R131 or FcγRIIa H131 -expressing HEK293T cells, the activation of CD40 after 24 h stimulation was indicated by the expression of GFP of the reporter cell and analyzed by flow cytometry. (B) B cells were stimulated by different concentrations of NK003 variants and the upregulation of CD23, CD86 or HLA-DR was analyzed by flow cytometry. MFI: Mean Fluorescence Intensity. (C) Schematic of OVA-specific CD8 + T cell response model. FcγR/CD40-humanized mice were adoptively transferred with OT-I cells, and then immunized with DEC-OVA in the presence of CD40 antibody variants, expansion of OT-I cells was analyzed by flow cytometry. (D) Quantification of OT-I cells (left) and the percentage of OT-I cells among CD8 + T cells (right) as in OVA-specific CD8 + T cell response model. Each circle represents an individual mouse. Bars represent the mean ± SEM. ** p ≤0.01, *** p ≤0.001, **** p ≤0.0001; One-way ANOVA Tukey's multiple comparisons test was used.

Techniques Used: Activity Assay, Expressing, Activation Assay, Flow Cytometry, Fluorescence, Mouse Assay

4) Product Images from "Laminin 411 acts as a potent inducer of umbilical cord mesenchymal stem cell differentiation into insulin-producing cells"

Article Title: Laminin 411 acts as a potent inducer of umbilical cord mesenchymal stem cell differentiation into insulin-producing cells

Journal: Journal of Translational Medicine

doi: 10.1186/1479-5876-12-135

UC-MSC phenotype analysis. MSCs derived from umbilical cord were detached with trypsin-EDTA and stained with fluorescence antibodies against surface molecules indicated and analyzed by flow cytometry present in histogram plots. Appropriate isotype controls were performed (for each antibody isotype) to assess cell auto-fluorescence and background staining. MSCs were positive for CD73 (99%), CD90 (99%), CD105 (95%), CD166 (98%), negative for hematopoietic marker CD34 (1.7%), leukocyte common antigen CD45 (2.2%), HLA-DR (1.3%) and monocyte marker CD14 (1.5%).
Figure Legend Snippet: UC-MSC phenotype analysis. MSCs derived from umbilical cord were detached with trypsin-EDTA and stained with fluorescence antibodies against surface molecules indicated and analyzed by flow cytometry present in histogram plots. Appropriate isotype controls were performed (for each antibody isotype) to assess cell auto-fluorescence and background staining. MSCs were positive for CD73 (99%), CD90 (99%), CD105 (95%), CD166 (98%), negative for hematopoietic marker CD34 (1.7%), leukocyte common antigen CD45 (2.2%), HLA-DR (1.3%) and monocyte marker CD14 (1.5%).

Techniques Used: Derivative Assay, Staining, Fluorescence, Flow Cytometry, Cytometry, Marker

5) Product Images from "Postoperative abdominal sepsis induces selective and persistent changes in CTCF binding within the MHC-II region of human monocytes"

Article Title: Postoperative abdominal sepsis induces selective and persistent changes in CTCF binding within the MHC-II region of human monocytes

Journal: PLoS ONE

doi: 10.1371/journal.pone.0250818

HLA-DR expression on monocyte surface. Surface expression of HLA-DR on CD14 ++ -monocytes at the time of sepsis diagnosis (T1) compared to matched control patients analyzed in whole blood samples using flow cytometry (p
Figure Legend Snippet: HLA-DR expression on monocyte surface. Surface expression of HLA-DR on CD14 ++ -monocytes at the time of sepsis diagnosis (T1) compared to matched control patients analyzed in whole blood samples using flow cytometry (p

Techniques Used: Expressing, Flow Cytometry

6) Product Images from "Human plasmacytoid dendritic cells at the crossroad of type I interferon-regulated B cell differentiation and antiviral response to tick-borne encephalitis virus"

Article Title: Human plasmacytoid dendritic cells at the crossroad of type I interferon-regulated B cell differentiation and antiviral response to tick-borne encephalitis virus

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1009505

Impact of TBEV E glycoprotein neutralization on IFN-α and Ig production. Total PBMC or isolated pDC were left untreated (NS) or stimulated with I-TBEV (dilution 1:12.5) alone or in combination either with two single-chain Ab (scAb) blocking TBEV E glycoprotein, B7 and G7 or with a scAb non-related to TBEV, A10 clone. (A) The release of IFN-α was measured by ELISA in culture supernatants of total PBMC left untreated (NS) or stimulated for 24 hours with I-TBEV alone or in combination either with B7 and G7 or with a non-related scAb A10 clone at the concentration of 0.15 μg/ml, 0.3 μg/ml and 0.6 μg/ml. The results shown were mean values ± standard error of the mean (SEM) of 3 independent experiments. ANOVA p value for IFN-α: 0.006. Based on LSD (equivalent to no adjustments). (B-D) Isolated pDC were left untreated (NS) or stimulated with I-TBEV alone or in combination either with G7 clone (0.6 μg/ml) or with A10 clone (0.6 μg/ml) for 24 hours. The production of IFN-α (B) was measured by ELISA in culture supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for IFN-α: 0.000. Based on LSD (equivalent to no adjustments). (C) pDC were stained with PD-L1, HLA-DR, CD86, CD80, ILT7, BDCA2 and BDCA4. A total of 50.000 cells were analyzed per sample by flow cytometry to evaluate the percentage of pDC sub-populations. A representative pDC sub-population profile out of 3 different experiments conducted separately is shown: P1-pDC (PD-L1 + CD80 - ) population is indicated in red while P2-pDC (PD-L1 + CD80 + ) in blue. TNF-α production (D) was tested by cytometric bead assay in 24 hour-collected supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for TNF-α: 0.002. Based on LSD (equivalent to no adjustments). (E-F) IgM and IgG production was measured by ELISA in supernatants collected from PBMC after 10 days of stimulation. The results are mean values ± SEM of 3 independent experiments for IgM and 2 independent experiments for IgG. ANOVA p value for IgM: 0.036. Based on LSD (equivalent to no adjustments).
Figure Legend Snippet: Impact of TBEV E glycoprotein neutralization on IFN-α and Ig production. Total PBMC or isolated pDC were left untreated (NS) or stimulated with I-TBEV (dilution 1:12.5) alone or in combination either with two single-chain Ab (scAb) blocking TBEV E glycoprotein, B7 and G7 or with a scAb non-related to TBEV, A10 clone. (A) The release of IFN-α was measured by ELISA in culture supernatants of total PBMC left untreated (NS) or stimulated for 24 hours with I-TBEV alone or in combination either with B7 and G7 or with a non-related scAb A10 clone at the concentration of 0.15 μg/ml, 0.3 μg/ml and 0.6 μg/ml. The results shown were mean values ± standard error of the mean (SEM) of 3 independent experiments. ANOVA p value for IFN-α: 0.006. Based on LSD (equivalent to no adjustments). (B-D) Isolated pDC were left untreated (NS) or stimulated with I-TBEV alone or in combination either with G7 clone (0.6 μg/ml) or with A10 clone (0.6 μg/ml) for 24 hours. The production of IFN-α (B) was measured by ELISA in culture supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for IFN-α: 0.000. Based on LSD (equivalent to no adjustments). (C) pDC were stained with PD-L1, HLA-DR, CD86, CD80, ILT7, BDCA2 and BDCA4. A total of 50.000 cells were analyzed per sample by flow cytometry to evaluate the percentage of pDC sub-populations. A representative pDC sub-population profile out of 3 different experiments conducted separately is shown: P1-pDC (PD-L1 + CD80 - ) population is indicated in red while P2-pDC (PD-L1 + CD80 + ) in blue. TNF-α production (D) was tested by cytometric bead assay in 24 hour-collected supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for TNF-α: 0.002. Based on LSD (equivalent to no adjustments). (E-F) IgM and IgG production was measured by ELISA in supernatants collected from PBMC after 10 days of stimulation. The results are mean values ± SEM of 3 independent experiments for IgM and 2 independent experiments for IgG. ANOVA p value for IgM: 0.036. Based on LSD (equivalent to no adjustments).

Techniques Used: Neutralization, Isolation, Blocking Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Staining, Flow Cytometry

Identification and characterization of cell population responsible for type I IFN production in PBMC. (A) Total PBMC or PBMC depleted of either pDC (PBMC-pDC) or monocytes (PBMC-Mo) were left untreated or stimulated with I-TBEV for 24 hours. The production of IFN-α was measured in culture supernatants by ELISA. The results shown were mean values ± SEM of 3 independent experiments. (B) PBMC were left untreated (NS) or stimulated with R848 (5 μM) or treated with I-TBEV (1:12.5) alone or in combination with the TLR-7/8 inhibitor, ODN 2087. The production of IFN-α was measured by ELISA in supernatants collected after 24 hours. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for IFN-α: 0.049. Based on LSD (equivalent to no adjustments). (C-F) Isolated pDC were left untreated (NS) or stimulated for 24 hours with I-TBEV alone or in combination with the TLR-7/8 inhibitor, ODN 2087. The production of IFN-α (C) was measured in culture supernatants by ELISA. The results shown are mean values ± SEM of 3 independent experiments. ANOVA p value for IFN-α: 0.000. Based on LSD (equivalent to no adjustments). (D) Isolated pDC were stained with PD-L1, HLA-DR, CD86, CD80, ILT7, BDCA2 and BDCA4. A total of 50.000 cells were analyzed per sample by flow cytometry to evaluate the percentage of pDC sub-populations in live BDCA4+ pDC. A representative pDC sub-population profile out of 3 different experiments conducted separately is shown: P1-pDC (PD-L1 + CD80 - ) population is indicated in red, P2-pDC (PD-L1 + CD80 + ) in blue. The production of TNF-α (E) and IL-6 (F) was tested by cytometric bead assay in 24 hour-collected supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for TNF-α: 0.003; for IL-6: 0.001. Based on LSD (equivalent to no adjustments).
Figure Legend Snippet: Identification and characterization of cell population responsible for type I IFN production in PBMC. (A) Total PBMC or PBMC depleted of either pDC (PBMC-pDC) or monocytes (PBMC-Mo) were left untreated or stimulated with I-TBEV for 24 hours. The production of IFN-α was measured in culture supernatants by ELISA. The results shown were mean values ± SEM of 3 independent experiments. (B) PBMC were left untreated (NS) or stimulated with R848 (5 μM) or treated with I-TBEV (1:12.5) alone or in combination with the TLR-7/8 inhibitor, ODN 2087. The production of IFN-α was measured by ELISA in supernatants collected after 24 hours. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for IFN-α: 0.049. Based on LSD (equivalent to no adjustments). (C-F) Isolated pDC were left untreated (NS) or stimulated for 24 hours with I-TBEV alone or in combination with the TLR-7/8 inhibitor, ODN 2087. The production of IFN-α (C) was measured in culture supernatants by ELISA. The results shown are mean values ± SEM of 3 independent experiments. ANOVA p value for IFN-α: 0.000. Based on LSD (equivalent to no adjustments). (D) Isolated pDC were stained with PD-L1, HLA-DR, CD86, CD80, ILT7, BDCA2 and BDCA4. A total of 50.000 cells were analyzed per sample by flow cytometry to evaluate the percentage of pDC sub-populations in live BDCA4+ pDC. A representative pDC sub-population profile out of 3 different experiments conducted separately is shown: P1-pDC (PD-L1 + CD80 - ) population is indicated in red, P2-pDC (PD-L1 + CD80 + ) in blue. The production of TNF-α (E) and IL-6 (F) was tested by cytometric bead assay in 24 hour-collected supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for TNF-α: 0.003; for IL-6: 0.001. Based on LSD (equivalent to no adjustments).

Techniques Used: Enzyme-linked Immunosorbent Assay, Isolation, Staining, Flow Cytometry

7) Product Images from "Autologous reconstitution of human cancer and immune system in vivo"

Article Title: Autologous reconstitution of human cancer and immune system in vivo

Journal: Oncotarget

doi: 10.18632/oncotarget.14026

Autologously reconstituted human carcinoma is infiltrated with endogenous human hematopoietic lymphocytes that mature in the thymus a. CD34+ BMC were transduced with lentivirus vectors containing GFP protein prior to BMC engraftment. Autologously reconstituted tumor was harvested, stained with DAPI, and immunofluorescent microscopy was performed. The left panels show tumor with only DAPI counterstain. Left upper panel is tumor xenografted into non-humanized mice, while the left lower panel is the tumor in autologously humanized mice (white arrowhead – lymphocyte; white arrow – myeloid cells). Slides were also co-stained with human CD3 and CD11b conjugates (right panels). The bar scale is 100µm. b. Human lymphocytes from humanized NOG-A2 (HLA-A0201 transgene) mice mature in the thymus. RBC depleted peripheral blood (PBL) from humanized mice were stained with hCD4 and hCD8. Thymus from these humanized mice were harvested and stained for hCD4 and hCD8. c. After tumors grew to 1 cm3 in autologously humanized mice, splenocytes and tumor were harvested for human HLA class I and class II typing. CD3 depletions were performed on the tumor specimens prior to this analysis. The haplotype report showed that human HLA class I and class II types between the tumor and the immune cells were identical. .
Figure Legend Snippet: Autologously reconstituted human carcinoma is infiltrated with endogenous human hematopoietic lymphocytes that mature in the thymus a. CD34+ BMC were transduced with lentivirus vectors containing GFP protein prior to BMC engraftment. Autologously reconstituted tumor was harvested, stained with DAPI, and immunofluorescent microscopy was performed. The left panels show tumor with only DAPI counterstain. Left upper panel is tumor xenografted into non-humanized mice, while the left lower panel is the tumor in autologously humanized mice (white arrowhead – lymphocyte; white arrow – myeloid cells). Slides were also co-stained with human CD3 and CD11b conjugates (right panels). The bar scale is 100µm. b. Human lymphocytes from humanized NOG-A2 (HLA-A0201 transgene) mice mature in the thymus. RBC depleted peripheral blood (PBL) from humanized mice were stained with hCD4 and hCD8. Thymus from these humanized mice were harvested and stained for hCD4 and hCD8. c. After tumors grew to 1 cm3 in autologously humanized mice, splenocytes and tumor were harvested for human HLA class I and class II typing. CD3 depletions were performed on the tumor specimens prior to this analysis. The haplotype report showed that human HLA class I and class II types between the tumor and the immune cells were identical. .

Techniques Used: Transduction, Staining, Microscopy, Mouse Assay

Autologously reconstituted HNSCC tumor system can be used to study human myeloid derived suppressor cells as well as novel immunomodulatory agents a. Splenocytes from autologously reconstituted human tumor bearing mice were harvested, and CD11b+CD14+DRlow (CD14) and CD11b+CD15+DRlow (CD15) cells were sorted to homogeneity to ensure monocytic and granulocytic histology (data not shown)[21]. Human CD3+ cells were co-cultured with CD11b+CD14+HLA-DRlow and CD11b+CD15+HLA-DRlow cells in complete RPMI media and stimulated (Sti) with PMA/Ionomycin. CD3+IFNγ+ cells were quantitated with flow cytometry as the measurement of T-cell suppression when mixed with MDSC. b. Palpable tumors of equivalent size in both groups were treated intratumorally with either PBS control or ML-RR-CDA at 20µg per injection and the tumor growth was followed. Experiment repeated twice with 4-5 mice per group. At the completion of experiment, the tumor was harvested and stained with human CD8, IFNγ, and DAPI counterstain (right panels). The bar scale is 100µ.
Figure Legend Snippet: Autologously reconstituted HNSCC tumor system can be used to study human myeloid derived suppressor cells as well as novel immunomodulatory agents a. Splenocytes from autologously reconstituted human tumor bearing mice were harvested, and CD11b+CD14+DRlow (CD14) and CD11b+CD15+DRlow (CD15) cells were sorted to homogeneity to ensure monocytic and granulocytic histology (data not shown)[21]. Human CD3+ cells were co-cultured with CD11b+CD14+HLA-DRlow and CD11b+CD15+HLA-DRlow cells in complete RPMI media and stimulated (Sti) with PMA/Ionomycin. CD3+IFNγ+ cells were quantitated with flow cytometry as the measurement of T-cell suppression when mixed with MDSC. b. Palpable tumors of equivalent size in both groups were treated intratumorally with either PBS control or ML-RR-CDA at 20µg per injection and the tumor growth was followed. Experiment repeated twice with 4-5 mice per group. At the completion of experiment, the tumor was harvested and stained with human CD8, IFNγ, and DAPI counterstain (right panels). The bar scale is 100µ.

Techniques Used: Derivative Assay, Mouse Assay, Cell Culture, Flow Cytometry, Injection, Staining

8) Product Images from "Elimination of Latently HIV-infected Cells from Antiretroviral Therapy-suppressed Subjects by Engineered Immune-mobilizing T-cell Receptors"

Article Title: Elimination of Latently HIV-infected Cells from Antiretroviral Therapy-suppressed Subjects by Engineered Immune-mobilizing T-cell Receptors

Journal: Molecular Therapy

doi: 10.1038/mt.2016.114

ImmTAVs mediate killing of infected CD4+ T-cells. ( a ) Confocal image of a conjugate between an HIV-infected CD4+ T-cell and healthy donor CD8+ T-cells in the presence of an ImmTAV (m121, 10 –9 mol/l). Red – p24 Ag; magenta – CD8. ( b ) Purified activated CD4+ T-cells from 5 HLA-A*0201-positive ART-treated patients were cultured with healthy donor CD8+ T-cells alone or with ImmTAVs at the concentrations indicated. Caspase-3 expression in CD4+ T-cells that were uninfected (left), HIV-infected singlets (middle) or HIV-infected and forming conjugates with CD8+ T-cells (right) was determined by flow cytometric analysis (gating strategy in Supplementary Figure S2 ). ( c ) Purified activated CD4+ T-cells from the same five ART-treated patients were cultured with healthy donor CD8+ T-cells (CD8+/CD4+ cell ratio = 1:1) alone or with an ImmTAV (m134, 10 –8 mol/l), which was either present throughout the culture period or washed out by replacing the culture medium after 48 hours. Reduction in Gag+ cells, normalized to CD8-only values, on day 7 of coculture is shown. In b and c , horizontal lines indicate mean values. ( d ) Schema (left) to illustrate the same coculture assay with NRTI-treated CD4+ T-cells. Tenofovir (10 µmol/l) was added to PHA-activated CD4+ T-cells from three ART-treated patients (006, 007, 008, black symbols, immediately after harvesting) and three healthy donors (HD1, HD2, HD3, gray symbols, immediately after spinoculation with HIV IIIB). After 48 hours of drug / mock drug exposure, healthy donor CD8+ T-cells and ImmTAVs were added and cultures were maintained for a further 48 hours. Percent elimination of HIV-infected cells normalized to no TCRs is shown (right). ART, ; PHA, ; HD, ; HIV, human immunodeficiency virus; ImmTAV, immune-mobilising monoclonal T-cell receptors against virus; TCR, T-cell receptor.
Figure Legend Snippet: ImmTAVs mediate killing of infected CD4+ T-cells. ( a ) Confocal image of a conjugate between an HIV-infected CD4+ T-cell and healthy donor CD8+ T-cells in the presence of an ImmTAV (m121, 10 –9 mol/l). Red – p24 Ag; magenta – CD8. ( b ) Purified activated CD4+ T-cells from 5 HLA-A*0201-positive ART-treated patients were cultured with healthy donor CD8+ T-cells alone or with ImmTAVs at the concentrations indicated. Caspase-3 expression in CD4+ T-cells that were uninfected (left), HIV-infected singlets (middle) or HIV-infected and forming conjugates with CD8+ T-cells (right) was determined by flow cytometric analysis (gating strategy in Supplementary Figure S2 ). ( c ) Purified activated CD4+ T-cells from the same five ART-treated patients were cultured with healthy donor CD8+ T-cells (CD8+/CD4+ cell ratio = 1:1) alone or with an ImmTAV (m134, 10 –8 mol/l), which was either present throughout the culture period or washed out by replacing the culture medium after 48 hours. Reduction in Gag+ cells, normalized to CD8-only values, on day 7 of coculture is shown. In b and c , horizontal lines indicate mean values. ( d ) Schema (left) to illustrate the same coculture assay with NRTI-treated CD4+ T-cells. Tenofovir (10 µmol/l) was added to PHA-activated CD4+ T-cells from three ART-treated patients (006, 007, 008, black symbols, immediately after harvesting) and three healthy donors (HD1, HD2, HD3, gray symbols, immediately after spinoculation with HIV IIIB). After 48 hours of drug / mock drug exposure, healthy donor CD8+ T-cells and ImmTAVs were added and cultures were maintained for a further 48 hours. Percent elimination of HIV-infected cells normalized to no TCRs is shown (right). ART, ; PHA, ; HD, ; HIV, human immunodeficiency virus; ImmTAV, immune-mobilising monoclonal T-cell receptors against virus; TCR, T-cell receptor.

Techniques Used: Infection, Purification, Cell Culture, Expressing, Flow Cytometry, Co-culture Assay

ImmTAV-mediated clearance of autologous HIV reservoir cells. ( a ). Schema illustrating assay to quantify ImmTAV-mediated elimination of resting HIV-infected Cd4+ T-cells from ART-treated patients after viral reactivation. CD25-/69-/HLA-DR- CD4+ cells were thoroughly washed after PHA treatment and cultured in duplicate at a density of 3 × 10 5 cells/well alone, with autologous CD8+ T-cells only or with healthy donor CD8+ T-cells plus ImmTAVs (10 –9 mol/l). Irradiated allogeneic PBMC feeders (1.5–3 × 10 6 ) were added to all wells. CD8+/CD4+ cell ratios were 1:1 throughout. After 14 days, Gag+ cells were quantified by flow cytometry. ( b ) Number of Gag+ cells remaining at day 14 of culture (mean of duplicate wells). ( c ) Percent elimination of Gag+ cells, determined by normalizing to no TCRs. In all subjects, culture supernatants from CD4+ cell-only wells were positive for free p24 Ag and ImmTAV-treated wells were negative (determined by enzyme-linked immunosorbent assay, cut-off 1500 pg/ml). (d) CD25-/69-/HLA-DR- CD4+ cells from five ART-treated HLA-A*0201-positive subjects were thoroughly washed after LRA (bryostatin / romidepsin) treatment and cultured in duplicate with healthy donor CD8+ T-cells (1:1 ratio) ± ImmTAV m121 or control TCR (10 –9 mol/l) for a further 42 hours. Supernatants were harvested and viral outgrowth was determined by quantification of HIV RNA, as described in Materials and Methods. LRA, latency-reversing agent; HLA, human histocompatibility leukocyte antigen; ART, antiretroviral therapy; PHA, phytohemagglutinin; HIV, human immunodeficiency virus; ImmTAV, immune-mobilising monoclonal T-cell receptors against virus; PBMC, peripheral blood mononuclear cells; TCR, T-cell receptor.
Figure Legend Snippet: ImmTAV-mediated clearance of autologous HIV reservoir cells. ( a ). Schema illustrating assay to quantify ImmTAV-mediated elimination of resting HIV-infected Cd4+ T-cells from ART-treated patients after viral reactivation. CD25-/69-/HLA-DR- CD4+ cells were thoroughly washed after PHA treatment and cultured in duplicate at a density of 3 × 10 5 cells/well alone, with autologous CD8+ T-cells only or with healthy donor CD8+ T-cells plus ImmTAVs (10 –9 mol/l). Irradiated allogeneic PBMC feeders (1.5–3 × 10 6 ) were added to all wells. CD8+/CD4+ cell ratios were 1:1 throughout. After 14 days, Gag+ cells were quantified by flow cytometry. ( b ) Number of Gag+ cells remaining at day 14 of culture (mean of duplicate wells). ( c ) Percent elimination of Gag+ cells, determined by normalizing to no TCRs. In all subjects, culture supernatants from CD4+ cell-only wells were positive for free p24 Ag and ImmTAV-treated wells were negative (determined by enzyme-linked immunosorbent assay, cut-off 1500 pg/ml). (d) CD25-/69-/HLA-DR- CD4+ cells from five ART-treated HLA-A*0201-positive subjects were thoroughly washed after LRA (bryostatin / romidepsin) treatment and cultured in duplicate with healthy donor CD8+ T-cells (1:1 ratio) ± ImmTAV m121 or control TCR (10 –9 mol/l) for a further 42 hours. Supernatants were harvested and viral outgrowth was determined by quantification of HIV RNA, as described in Materials and Methods. LRA, latency-reversing agent; HLA, human histocompatibility leukocyte antigen; ART, antiretroviral therapy; PHA, phytohemagglutinin; HIV, human immunodeficiency virus; ImmTAV, immune-mobilising monoclonal T-cell receptors against virus; PBMC, peripheral blood mononuclear cells; TCR, T-cell receptor.

Techniques Used: Infection, Cell Culture, Irradiation, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

ImmTAV-mediated elimination of activated and resting HIV-infected CD4+ T-cells. Purified PHA-stimulated CD4+ T-cells from the same five HLA-A*0201-positive ART-treated patients as in Figure 5 were cultured with healthy donor CD8+ T-cells alone or with ImmTAV m134 (10 –8 mol/l). The ImmTAV was either present for the duration of coculture or washed out after 48 hours. ( a ) The number of activated (CD25+/CD69+/HLA-DR+) and resting (CD25-/CD69-/HLA-DR-) Gag+ CD4+ T-cells remaining on day 7 of coculture and ( b ) percent elimination of infected cells, normalized to CD8-only values, within each subset is shown. ( c ) Mean fluorescence intensity (MFI, arbitrary units) of Gag expression in residual infected activated and resting CD4+ T-cells after culture under the conditions indicated (as for a and b ; bars and horizontal lines represent range and mean respectively). Consistency in MFI values was ensured by acquiring all samples in a single run. ( d ). Correlation between Gag MFI (arbitrary units) and normalized percent elimination of infected cells. ( e ) Biotinylated m134 ImmTAV was detected by microscopy on the surface of individual HIV-1 IIIB-infected CD4+ T-cells from two healthy donors after staining with streptavidin-PE. Data shown in left panel represent counts of fluorescent spots; the total for each cell was obtained using Z-stack images. Uninfected cells were used to determine background staining. Right panel: representative phase-contrast (top) and corresponding fluorescence images (bottom) of infected CD4+ T-cells from donor 1 stained with m134 ImmTAV. Fluorescence images are 3D reconstructions of individual planes. The brightness/contrast of images was adjusted to optimize epitope visualization. The scale bar represents 10 µm. HLA, human histocompatibility leukocyte antigen; PHA, phytohemagglutinin; HIV, human immunodeficiency virus; ImmTAV, immune-mobilising monoclonal T-cell receptors against virus.
Figure Legend Snippet: ImmTAV-mediated elimination of activated and resting HIV-infected CD4+ T-cells. Purified PHA-stimulated CD4+ T-cells from the same five HLA-A*0201-positive ART-treated patients as in Figure 5 were cultured with healthy donor CD8+ T-cells alone or with ImmTAV m134 (10 –8 mol/l). The ImmTAV was either present for the duration of coculture or washed out after 48 hours. ( a ) The number of activated (CD25+/CD69+/HLA-DR+) and resting (CD25-/CD69-/HLA-DR-) Gag+ CD4+ T-cells remaining on day 7 of coculture and ( b ) percent elimination of infected cells, normalized to CD8-only values, within each subset is shown. ( c ) Mean fluorescence intensity (MFI, arbitrary units) of Gag expression in residual infected activated and resting CD4+ T-cells after culture under the conditions indicated (as for a and b ; bars and horizontal lines represent range and mean respectively). Consistency in MFI values was ensured by acquiring all samples in a single run. ( d ). Correlation between Gag MFI (arbitrary units) and normalized percent elimination of infected cells. ( e ) Biotinylated m134 ImmTAV was detected by microscopy on the surface of individual HIV-1 IIIB-infected CD4+ T-cells from two healthy donors after staining with streptavidin-PE. Data shown in left panel represent counts of fluorescent spots; the total for each cell was obtained using Z-stack images. Uninfected cells were used to determine background staining. Right panel: representative phase-contrast (top) and corresponding fluorescence images (bottom) of infected CD4+ T-cells from donor 1 stained with m134 ImmTAV. Fluorescence images are 3D reconstructions of individual planes. The brightness/contrast of images was adjusted to optimize epitope visualization. The scale bar represents 10 µm. HLA, human histocompatibility leukocyte antigen; PHA, phytohemagglutinin; HIV, human immunodeficiency virus; ImmTAV, immune-mobilising monoclonal T-cell receptors against virus.

Techniques Used: Infection, Purification, Cell Culture, Fluorescence, Expressing, Microscopy, Staining

9) Product Images from "STING agonist formulated cancer vaccines can cure established tumors resistant to PD-1 blockade"

Article Title: STING agonist formulated cancer vaccines can cure established tumors resistant to PD-1 blockade

Journal: Science translational medicine

doi: 10.1126/scitranslmed.aaa4306

ML–RR-S2–CDA potently activates human APCs ( A ) Human THP1-Blue cells with IRF3 reporter gene were stimulated with 50 μM of CDNs, and the concentration of secreted embryonic alkaline phosphatase (SEAP) reporter was measured by spectrophotometry. Histogram represents mean ± SD of triplicate samples. ( B ) PBMCs from a WT STING human donor were stimulated with 50 μM of the indicated CDNs. After 24 hours, intracellular IFNα on gated CD14 + monocytes or on CD11c + HLA-DR + DCs (mDCs) was measured by fluorescence-activated cell sorting (FACS). ( C ) Cultured human DCs were stimulated with 50 μM synthetic CDNs or LPS (1 μg/ml) as a positive control. After 48 hours, MHCI (HLA-ABC), CD80, CD83, and CD86 were measured on the gated CD11c + DCs. Bar graphs indicate the average MFI (top panel); representative histograms show CD80, CD86, CD83, and MHCI expression in human DCs. Filled histograms correspond to unstimulated cells, the dotted line represents LPS stimulation, and the solid line represents stimulation with ML–RR-S2–CDA (bottom). ( D ) Synthetic noncanonical CDNs can stimulate all the hSTING alleles with increased potency. Human PBMCs from four donors, each with a different STING genotype (STING WT , STING HAQ , STING WT/REF , or STING HAQ/REF ), were stimulated with 10 μM of the indicated CDNs. After 6 hours of incubation, the supernatants were harvested for analysis of TNFα protein (top), and the cells were harvested for analysis of IFN β induction by qRT-PCR (bottom). ( E ) CDN-stimulated DCs can activate T H 1 response in human T cells. Human DCs (CD11c + cells) were probed for IL-12 expression after treatment with GM-CSF, LPS, or ML–RR-S2–CDA. Human DCs treated with GM-CSF, LPS, or ML–RR-S2–CDA were used in MLRs to stimulate human T cells (gated for CD8 + cells). Intracellular IFNγ was measured in CD8 + -gated cells. Left panels display mean MFI (IL-12, left; IFNγ, right), and right panels show representative histograms from gated flow analysis. Data are means ± SD of five samples.
Figure Legend Snippet: ML–RR-S2–CDA potently activates human APCs ( A ) Human THP1-Blue cells with IRF3 reporter gene were stimulated with 50 μM of CDNs, and the concentration of secreted embryonic alkaline phosphatase (SEAP) reporter was measured by spectrophotometry. Histogram represents mean ± SD of triplicate samples. ( B ) PBMCs from a WT STING human donor were stimulated with 50 μM of the indicated CDNs. After 24 hours, intracellular IFNα on gated CD14 + monocytes or on CD11c + HLA-DR + DCs (mDCs) was measured by fluorescence-activated cell sorting (FACS). ( C ) Cultured human DCs were stimulated with 50 μM synthetic CDNs or LPS (1 μg/ml) as a positive control. After 48 hours, MHCI (HLA-ABC), CD80, CD83, and CD86 were measured on the gated CD11c + DCs. Bar graphs indicate the average MFI (top panel); representative histograms show CD80, CD86, CD83, and MHCI expression in human DCs. Filled histograms correspond to unstimulated cells, the dotted line represents LPS stimulation, and the solid line represents stimulation with ML–RR-S2–CDA (bottom). ( D ) Synthetic noncanonical CDNs can stimulate all the hSTING alleles with increased potency. Human PBMCs from four donors, each with a different STING genotype (STING WT , STING HAQ , STING WT/REF , or STING HAQ/REF ), were stimulated with 10 μM of the indicated CDNs. After 6 hours of incubation, the supernatants were harvested for analysis of TNFα protein (top), and the cells were harvested for analysis of IFN β induction by qRT-PCR (bottom). ( E ) CDN-stimulated DCs can activate T H 1 response in human T cells. Human DCs (CD11c + cells) were probed for IL-12 expression after treatment with GM-CSF, LPS, or ML–RR-S2–CDA. Human DCs treated with GM-CSF, LPS, or ML–RR-S2–CDA were used in MLRs to stimulate human T cells (gated for CD8 + cells). Intracellular IFNγ was measured in CD8 + -gated cells. Left panels display mean MFI (IL-12, left; IFNγ, right), and right panels show representative histograms from gated flow analysis. Data are means ± SD of five samples.

Techniques Used: Concentration Assay, Spectrophotometry, Fluorescence, FACS, Cell Culture, Positive Control, Expressing, Incubation, Quantitative RT-PCR, Flow Cytometry

10) Product Images from "Gene profiling analysis of ALVAC infected human monocyte derived dendritic cells"

Article Title: Gene profiling analysis of ALVAC infected human monocyte derived dendritic cells

Journal: Vaccine

doi: 10.1016/j.vaccine.2008.07.050

Maturation of human immature MDDCs upon exposure to ALVAC. (A) Human immature MDDCs (CD11c+ and CD14−), obtained from CD14 purified monocytes upon 6-day culture with GM-CSF and IL-4, were stimulated for 24 h with ALVAC (0.2 MOI), LPS (10 ng/ml) or left untreated (indicated by color code). (B) Plots show super-imposition of the surface expression of each maturation marker relative to the Ab isotype control under the three experimental conditions (ALVAC, LPS or medium). ALVAC infection increased the expression of DC maturation-associated markers CD40, CD80, CD86, CD83, HLA-1 (MHC-I), HLA-DR (MHC-II) on the surface of human MDDC. However, in contrast to LPS, ALVAC failed to up-regulate the CD25 marker.
Figure Legend Snippet: Maturation of human immature MDDCs upon exposure to ALVAC. (A) Human immature MDDCs (CD11c+ and CD14−), obtained from CD14 purified monocytes upon 6-day culture with GM-CSF and IL-4, were stimulated for 24 h with ALVAC (0.2 MOI), LPS (10 ng/ml) or left untreated (indicated by color code). (B) Plots show super-imposition of the surface expression of each maturation marker relative to the Ab isotype control under the three experimental conditions (ALVAC, LPS or medium). ALVAC infection increased the expression of DC maturation-associated markers CD40, CD80, CD86, CD83, HLA-1 (MHC-I), HLA-DR (MHC-II) on the surface of human MDDC. However, in contrast to LPS, ALVAC failed to up-regulate the CD25 marker.

Techniques Used: Purification, Expressing, Marker, Infection

11) Product Images from "Granulocyte-colony stimulating factor drives the in vitro differentiation of human dendritic cells that induce anergy in na?ve T cells"

Article Title: Granulocyte-colony stimulating factor drives the in vitro differentiation of human dendritic cells that induce anergy in na?ve T cells

Journal: European Journal of Immunology

doi: 10.1002/eji.201040659

G-CSF drives the differentiation of DC-like cells similar to post-G DC. MoDC were differentiated by 7-day culture in the presence of G-CSF and IL-4 (G-DC), or GM-CSF and IL-4 (iDC), in some cases with the addition of LPS during the last 2 days of culture (mDC). (A) Cytospins were performed by centrifugation of 10 5 cells onto slides, followed by staining with May Grünwald-Giemsa (magnification 60×). One representative donor out of four tested in two independent experiments is shown. (B–D) Expression of CD14, CD1a, CD16 (B), HLA-DR, CD80, CD83, CD86 (B and C), ILT2, ILT3, ILT4 and HLA-G (D) was evaluated by FACS on day 7. A representative donor and the mean+SEM of at least ten donors tested in five independent experiments are presented. * p
Figure Legend Snippet: G-CSF drives the differentiation of DC-like cells similar to post-G DC. MoDC were differentiated by 7-day culture in the presence of G-CSF and IL-4 (G-DC), or GM-CSF and IL-4 (iDC), in some cases with the addition of LPS during the last 2 days of culture (mDC). (A) Cytospins were performed by centrifugation of 10 5 cells onto slides, followed by staining with May Grünwald-Giemsa (magnification 60×). One representative donor out of four tested in two independent experiments is shown. (B–D) Expression of CD14, CD1a, CD16 (B), HLA-DR, CD80, CD83, CD86 (B and C), ILT2, ILT3, ILT4 and HLA-G (D) was evaluated by FACS on day 7. A representative donor and the mean+SEM of at least ten donors tested in five independent experiments are presented. * p

Techniques Used: Centrifugation, Staining, Expressing, FACS

12) Product Images from "Immune Response to the West Nile Virus in Aged Non-Human Primates"

Article Title: Immune Response to the West Nile Virus in Aged Non-Human Primates

Journal: PLoS ONE

doi: 10.1371/journal.pone.0015514

B cells Proliferate in Response to WNV Infection. Panel A and B: Representative profiles from flow cytometric analysis of PBMC from animals in Cohort 1 and Cohort 2 prior to and at day 10 post WNV infection. The first panel illustrates the physical size and cellular granularity (forward side scatter profile) of the PBMC. The next series of panels illustrate the sub populations defined first by expression of CD20 (to define B cells) and CD3 status (T cells vs non-T cells). The next series illustrates the proliferation (Ki67+) within the B cells (CD3- CD20+ HLA-DR+). The final calculations are expressed as percentages based upon the CD3- population for the B cells. Panel B illustrates the differences in B cell proliferation found in Group 1 vs. Group 2 at d10 post infection. Lower panels (C and D) illustrate B cell proliferation (CD3-CD20+HLA/DR+ based upon levels of Ki67 measured within the total CD3- populations for cohort 1 and 2. Panels C and D illustrate proliferation of Ki67+CD20+ cells in cohorts 1 and 2 respectively,. Dark lines illustrate the aged animals lighter lines illustrate the adult animals.
Figure Legend Snippet: B cells Proliferate in Response to WNV Infection. Panel A and B: Representative profiles from flow cytometric analysis of PBMC from animals in Cohort 1 and Cohort 2 prior to and at day 10 post WNV infection. The first panel illustrates the physical size and cellular granularity (forward side scatter profile) of the PBMC. The next series of panels illustrate the sub populations defined first by expression of CD20 (to define B cells) and CD3 status (T cells vs non-T cells). The next series illustrates the proliferation (Ki67+) within the B cells (CD3- CD20+ HLA-DR+). The final calculations are expressed as percentages based upon the CD3- population for the B cells. Panel B illustrates the differences in B cell proliferation found in Group 1 vs. Group 2 at d10 post infection. Lower panels (C and D) illustrate B cell proliferation (CD3-CD20+HLA/DR+ based upon levels of Ki67 measured within the total CD3- populations for cohort 1 and 2. Panels C and D illustrate proliferation of Ki67+CD20+ cells in cohorts 1 and 2 respectively,. Dark lines illustrate the aged animals lighter lines illustrate the adult animals.

Techniques Used: Infection, Flow Cytometry, Expressing

13) Product Images from "Evaluation of the Effectiveness of Mesenchymal Stem Cells of the Placenta and Their Conditioned Medium in Local Radiation Injuries"

Article Title: Evaluation of the Effectiveness of Mesenchymal Stem Cells of the Placenta and Their Conditioned Medium in Local Radiation Injuries

Journal: Cells

doi: 10.3390/cells9122558

MSCs derived from the placenta immunophenotype: CD90+/CD105+73+/CD45−/CD34−/HLA-DR−, 7−ADD (99.5%).
Figure Legend Snippet: MSCs derived from the placenta immunophenotype: CD90+/CD105+73+/CD45−/CD34−/HLA-DR−, 7−ADD (99.5%).

Techniques Used: Derivative Assay

14) Product Images from "CD80 on Human T Cells Is Associated With FoxP3 Expression and Supports Treg Homeostasis"

Article Title: CD80 on Human T Cells Is Associated With FoxP3 Expression and Supports Treg Homeostasis

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2020.577655

CD80 and CD86 are expressed on human in vivo activated conventional and regulatory T cells. (A) Graphs show percentage of CD80 + and CD86 + T cells across seven donors. (B) Cells were gated on CD3 + CD4 + T cells and based on FoxP3 expression divided into Treg and Tcon. Cells were further gated on Ki-67 and analysed for CD80 and CD86 expression. (C) Graphs show percentage of CD80 + and CD86 + Tcon and Treg expressing either high or low levels of Ki-67, CTLA-4, HLA-DR and CD45-RA. (D , E) Boxplots show CD80 and CD86 RNA expression levels (D) and promoter activity (E) . Data were downloaded from DICE and FANTOM5 databases respectively. TPM—transcripts per million. All p-values were calculated using unpaired two-tailed T-test.
Figure Legend Snippet: CD80 and CD86 are expressed on human in vivo activated conventional and regulatory T cells. (A) Graphs show percentage of CD80 + and CD86 + T cells across seven donors. (B) Cells were gated on CD3 + CD4 + T cells and based on FoxP3 expression divided into Treg and Tcon. Cells were further gated on Ki-67 and analysed for CD80 and CD86 expression. (C) Graphs show percentage of CD80 + and CD86 + Tcon and Treg expressing either high or low levels of Ki-67, CTLA-4, HLA-DR and CD45-RA. (D , E) Boxplots show CD80 and CD86 RNA expression levels (D) and promoter activity (E) . Data were downloaded from DICE and FANTOM5 databases respectively. TPM—transcripts per million. All p-values were calculated using unpaired two-tailed T-test.

Techniques Used: In Vivo, Expressing, RNA Expression, Activity Assay, Two Tailed Test

15) Product Images from "Signal peptide cleavage is essential for surface expression of a regulatory T cell surface protein, leucine rich repeat containing 32 (LRRC32)"

Article Title: Signal peptide cleavage is essential for surface expression of a regulatory T cell surface protein, leucine rich repeat containing 32 (LRRC32)

Journal: BMC Biochemistry

doi: 10.1186/1471-2091-12-27

LRRC32 + CD4 + CD25 hi FoxP3 T regs appear to be more potent suppressors than LRRC32 - CD4 + CD25 hi FoxP3 and exhibit decreased CD62L upon activation . a) Expression of LRRC32 and LAP in CD4+ T cells rested overnight (top panel) or stimulated with plate bound anti-CD3 and soluble anti-CD28 (bottom panel). T regs were selected from the top 5% CD25-expressing and FoxP3 + populations, as previously described. Confirmation of activation by expression of the surface markers CD40L and CD69 are also shown (top of each panel). b) The expression patterns of various T reg and activation surface markers (CD62L, CD69, GITR, CTLA4, CD45RO, and HLA-DR) in FoxP3 + and LRRC32 + -gated populations of CD25 hi cells were studied using flow cytometry. Stimulated CD4 + FoxP3 + CD25 hi T regs (top panel) unstimulated CD4 + FoxP3 + CD25 hi T regs (bottom panel). c) Composite summary of phenotypic analysis of unstimulated LRRC32 - CD4 + CD25 hi FoxP3 + T regs and stimulated LRRC32 + and LRRC32 - CD4 + CD25 hi FoxP3 + T regs . Black bars = unstimulated LRRC32 - T regs . Dark grey bars = stimulated LRRC32 - T regs . Light grey bars = stimulated LRRC32 + T regs . Data are expressed as the mean ± SEM from 3 individuals. Heteroscedastic variances and an independent t-test comparing stimulated LRRC32 + and LRRC32 - subsets were used for calculations of the p values which are reported along the x-axis, below each surface marker (*). d) CD25 hi cells were sorted and activated overnight using anti-CD3-coated plates and soluble anti-CD28 (1 microgram/ml). Cells were then resorted based upon LRRC32 expression. The suppressive capacities of these LRRC32 + and LRRC32 - T regs were subsequently tested in a mixed lymphocyte reaction utilizing syngeneic effectors (T eff , 20,000/well) and allogenic antigen presenting cells (50,000/well). T reg :T eff ratios are depicted above. Data summarize 3 independent experiments. Results are expressed as the mean ± SEM. p = 0.0001 and R 2 = 0.7244. Absolute proliferation values for the 3 experiments were as follow: T effs alone: average of 31094 cpm to average of 47483 cpm (at least 6 replicates per assay), background: average of 24 cpm to 35 cpm (at least 6 replicates per assay); T reg :T eff ratio of 1:1: 89 cpm to 346 cpm. When titrating T regs vs. T effs , 3 replicates were performed at each titration for the LRRC32 + and LRRC32 - T regs except for in one assay set in which there was limited number of LRRC32 + T regs . In this case, only one replicate was performed at the 1:1 and 1:2 titrations, and two replicates were performed for the other titrations (0:1, 1:4, 1:8, and 1:16). We performed 3 replicates for each titration utilizing the LRRC32 - T regs .
Figure Legend Snippet: LRRC32 + CD4 + CD25 hi FoxP3 T regs appear to be more potent suppressors than LRRC32 - CD4 + CD25 hi FoxP3 and exhibit decreased CD62L upon activation . a) Expression of LRRC32 and LAP in CD4+ T cells rested overnight (top panel) or stimulated with plate bound anti-CD3 and soluble anti-CD28 (bottom panel). T regs were selected from the top 5% CD25-expressing and FoxP3 + populations, as previously described. Confirmation of activation by expression of the surface markers CD40L and CD69 are also shown (top of each panel). b) The expression patterns of various T reg and activation surface markers (CD62L, CD69, GITR, CTLA4, CD45RO, and HLA-DR) in FoxP3 + and LRRC32 + -gated populations of CD25 hi cells were studied using flow cytometry. Stimulated CD4 + FoxP3 + CD25 hi T regs (top panel) unstimulated CD4 + FoxP3 + CD25 hi T regs (bottom panel). c) Composite summary of phenotypic analysis of unstimulated LRRC32 - CD4 + CD25 hi FoxP3 + T regs and stimulated LRRC32 + and LRRC32 - CD4 + CD25 hi FoxP3 + T regs . Black bars = unstimulated LRRC32 - T regs . Dark grey bars = stimulated LRRC32 - T regs . Light grey bars = stimulated LRRC32 + T regs . Data are expressed as the mean ± SEM from 3 individuals. Heteroscedastic variances and an independent t-test comparing stimulated LRRC32 + and LRRC32 - subsets were used for calculations of the p values which are reported along the x-axis, below each surface marker (*). d) CD25 hi cells were sorted and activated overnight using anti-CD3-coated plates and soluble anti-CD28 (1 microgram/ml). Cells were then resorted based upon LRRC32 expression. The suppressive capacities of these LRRC32 + and LRRC32 - T regs were subsequently tested in a mixed lymphocyte reaction utilizing syngeneic effectors (T eff , 20,000/well) and allogenic antigen presenting cells (50,000/well). T reg :T eff ratios are depicted above. Data summarize 3 independent experiments. Results are expressed as the mean ± SEM. p = 0.0001 and R 2 = 0.7244. Absolute proliferation values for the 3 experiments were as follow: T effs alone: average of 31094 cpm to average of 47483 cpm (at least 6 replicates per assay), background: average of 24 cpm to 35 cpm (at least 6 replicates per assay); T reg :T eff ratio of 1:1: 89 cpm to 346 cpm. When titrating T regs vs. T effs , 3 replicates were performed at each titration for the LRRC32 + and LRRC32 - T regs except for in one assay set in which there was limited number of LRRC32 + T regs . In this case, only one replicate was performed at the 1:1 and 1:2 titrations, and two replicates were performed for the other titrations (0:1, 1:4, 1:8, and 1:16). We performed 3 replicates for each titration utilizing the LRRC32 - T regs .

Techniques Used: Activation Assay, Expressing, Flow Cytometry, Marker, Titration

16) Product Images from "Engineered mucoperiosteal scaffold for cleft palate regeneration towards the non-immunogenic transplantation"

Article Title: Engineered mucoperiosteal scaffold for cleft palate regeneration towards the non-immunogenic transplantation

Journal: Scientific Reports

doi: 10.1038/s41598-021-93951-w

( a – d ) MSCs characterization after in vitro expansion. ( a ) Cells, isolated from the bone marrow of a healthy donor, display the typical spindle-shaped morphology of MSCs. ( b ) Immunophenotyping by flow cytometry reveals expression of MSCs markers (CD73, CD90, and CD105) and lack of hematopoietic markers (CD34, CD45, CD80, CD86, and HLA-DR). ( c ) After incubation with the appropriate medium, the isolated cells were able to differentiate into osteoblasts as revealed by Alizarin Red staining (Sigma-Aldrich), which highlights calcium depositions. ( d ) At the same time, the isolated cells were also able to differentiate into adipocytes, as detected by Oil Red O (Sigma-Aldrich), which makes fat droplets visible. All the figures in the panel are the same scale; bar—200 µm.
Figure Legend Snippet: ( a – d ) MSCs characterization after in vitro expansion. ( a ) Cells, isolated from the bone marrow of a healthy donor, display the typical spindle-shaped morphology of MSCs. ( b ) Immunophenotyping by flow cytometry reveals expression of MSCs markers (CD73, CD90, and CD105) and lack of hematopoietic markers (CD34, CD45, CD80, CD86, and HLA-DR). ( c ) After incubation with the appropriate medium, the isolated cells were able to differentiate into osteoblasts as revealed by Alizarin Red staining (Sigma-Aldrich), which highlights calcium depositions. ( d ) At the same time, the isolated cells were also able to differentiate into adipocytes, as detected by Oil Red O (Sigma-Aldrich), which makes fat droplets visible. All the figures in the panel are the same scale; bar—200 µm.

Techniques Used: In Vitro, Isolation, Flow Cytometry, Expressing, Incubation, Staining

17) Product Images from "BDCA-2, a Novel Plasmacytoid Dendritic Cell-specific Type II C-type Lectin, Mediates Antigen Capture and Is a Potent Inhibitor of Interferon ?/? Induction"

Article Title: BDCA-2, a Novel Plasmacytoid Dendritic Cell-specific Type II C-type Lectin, Mediates Antigen Capture and Is a Potent Inhibitor of Interferon ?/? Induction

Journal: The Journal of Experimental Medicine

doi:

Anatomical localization of BDCA-2–expressing PDCs in inflamed tonsils. Fluorescent double staining with FITC–conjugated (green) anti–BDCA-2 mAb and Texas Red–conjugated (red) CD8 mAb (A), CD20 mAb (B), CD123 mAb (C), and anti–HLA-DR mAb (D). Note that BDCA-2–expressing PDCs are found in the T cell-rich extrafollicular areas but not within the germinal center. Like CD123, BDCA-2 is expressed on PDCs, but unlike CD123, BDCA-2 is not expressed on HEVs. One representative experiment of three is shown.
Figure Legend Snippet: Anatomical localization of BDCA-2–expressing PDCs in inflamed tonsils. Fluorescent double staining with FITC–conjugated (green) anti–BDCA-2 mAb and Texas Red–conjugated (red) CD8 mAb (A), CD20 mAb (B), CD123 mAb (C), and anti–HLA-DR mAb (D). Note that BDCA-2–expressing PDCs are found in the T cell-rich extrafollicular areas but not within the germinal center. Like CD123, BDCA-2 is expressed on PDCs, but unlike CD123, BDCA-2 is not expressed on HEVs. One representative experiment of three is shown.

Techniques Used: Expressing, Double Staining

18) Product Images from "Calcium Signaling through Phospholipase C Activates Dendritic Cells To Mature and Is Necessary for the Activation and Maturation of Dendritic Cells Induced by Diverse Agonists"

Article Title: Calcium Signaling through Phospholipase C Activates Dendritic Cells To Mature and Is Necessary for the Activation and Maturation of Dendritic Cells Induced by Diverse Agonists

Journal: Clinical and Diagnostic Laboratory Immunology

doi: 10.1128/CDLI.11.1.77-82.2004

PMT activates MDDCs to mature. Cell surface expression of the indicated markers on untreated MDDCs (dotted histograms) or MDDCs treated with the indicated agonists (solid histograms) is shown. Day 4 MDDCs were incubated with the indicated concentrations of PMT for 20 h. The cells were then harvested and stained for four-color flow cytometry with phycoerythrin-labeled anti-CD80, fluorescein isothiocyanate-labeled anti-CD83, cytochrome-labeled anti-CD86, and anti-HLA-DR APCs. The percentage of cells that increased the expression of the indicated markers (percent activation [the numbers in the histograms]) was calculated as described in Materials and Methods. The data shown are from one of six experiments performed, with similar results obtained in each experiment.
Figure Legend Snippet: PMT activates MDDCs to mature. Cell surface expression of the indicated markers on untreated MDDCs (dotted histograms) or MDDCs treated with the indicated agonists (solid histograms) is shown. Day 4 MDDCs were incubated with the indicated concentrations of PMT for 20 h. The cells were then harvested and stained for four-color flow cytometry with phycoerythrin-labeled anti-CD80, fluorescein isothiocyanate-labeled anti-CD83, cytochrome-labeled anti-CD86, and anti-HLA-DR APCs. The percentage of cells that increased the expression of the indicated markers (percent activation [the numbers in the histograms]) was calculated as described in Materials and Methods. The data shown are from one of six experiments performed, with similar results obtained in each experiment.

Techniques Used: Expressing, Incubation, Staining, Flow Cytometry, Cytometry, Labeling, Activation Assay

19) Product Images from "Potential for virus endogenization in humans through testicular germ cell infection: the case of HIV"

Article Title: Potential for virus endogenization in humans through testicular germ cell infection: the case of HIV

Journal: bioRxiv

doi: 10.1101/2020.06.04.135657

HIV-1 integration in testicular germ cell genome and in vivo detection of HIV and SIV DNA in human and non-human primate testicular germ cells ( A-C ) HIV-1 integration in primary TGCs. ( A ) Experimental design: after overnight contact with Jurkat cells infected with HIV-1 nef -ires-GFP, TGCs were recovered following CD45 magnetic selection and put in culture for 5 days. The cells were then submitted to FACS sorting for live cells, negative for leukocytes markers CD45, CD3 and HLA-DR and positive for germ cell markers DDX4 and MAGEA4. As a negative control, TGCs purified by CD45 magnetic selection immediately after contact with Jurkat cells were sorted by FACS similarly to the overnight incubated samples. ( B ) Gating strategy and representative profile of purified live germ cells. Live single cells were selected based on absence of detection of leukocytes markers CD3, HLA-DR and CD45 and on positive expression of DDX4 and MageA4+ germ cell markers. Control antibodies are shown on bottom panel. ( C ) HIV-1 integrated DNA was measured by Alu-gag PCR on 10,000 sorted TGCs exposed ON to Jurkat cells in 3 independent experiments, and the value obtained for the negative controls subtracted. ( D-F ) HIV integration in Tcam-2. ( D ) HIV-1 integrated DNA was measured on Tcam-2 either cultured for 48h following exposure to R5 JR-CSF HIV-1 (cell-free) or exposed ON to Jurkat infected with HIV R5 JR-CSF , purified by CD45 magnetic selection, cultured for 5 days and FACS sorted for CD45 negative live cells (cell associated). ( E, F ) HIV-1 reverse transcription and integration were assessed on Tcam-2 cells exposed to VSV-G-pseudotyped HIV-1. ( E ) Reverse transcripts were detected after 24h culture with or without reverse transcriptase inhibitor nevirapine (n=6) and ( F ) HIV-1 integration assessed by Alu-gag PCR (n=3). ( G ) In vivo detection of HIV and SIV DNA in human and non-human primate testicular germ cells by DNAscope. Representative pictures of TGC (green) harbouring viral DNA (red) within testis tissue section from one HIV-1 infected man (VL=51 681 cp/mL), one chronically infected rhesus macaque (VL > 10^6 cp/mL) and one African Green monkey 64 days post infection (VL=12085 cp/mL). Scale bars= 100μm.
Figure Legend Snippet: HIV-1 integration in testicular germ cell genome and in vivo detection of HIV and SIV DNA in human and non-human primate testicular germ cells ( A-C ) HIV-1 integration in primary TGCs. ( A ) Experimental design: after overnight contact with Jurkat cells infected with HIV-1 nef -ires-GFP, TGCs were recovered following CD45 magnetic selection and put in culture for 5 days. The cells were then submitted to FACS sorting for live cells, negative for leukocytes markers CD45, CD3 and HLA-DR and positive for germ cell markers DDX4 and MAGEA4. As a negative control, TGCs purified by CD45 magnetic selection immediately after contact with Jurkat cells were sorted by FACS similarly to the overnight incubated samples. ( B ) Gating strategy and representative profile of purified live germ cells. Live single cells were selected based on absence of detection of leukocytes markers CD3, HLA-DR and CD45 and on positive expression of DDX4 and MageA4+ germ cell markers. Control antibodies are shown on bottom panel. ( C ) HIV-1 integrated DNA was measured by Alu-gag PCR on 10,000 sorted TGCs exposed ON to Jurkat cells in 3 independent experiments, and the value obtained for the negative controls subtracted. ( D-F ) HIV integration in Tcam-2. ( D ) HIV-1 integrated DNA was measured on Tcam-2 either cultured for 48h following exposure to R5 JR-CSF HIV-1 (cell-free) or exposed ON to Jurkat infected with HIV R5 JR-CSF , purified by CD45 magnetic selection, cultured for 5 days and FACS sorted for CD45 negative live cells (cell associated). ( E, F ) HIV-1 reverse transcription and integration were assessed on Tcam-2 cells exposed to VSV-G-pseudotyped HIV-1. ( E ) Reverse transcripts were detected after 24h culture with or without reverse transcriptase inhibitor nevirapine (n=6) and ( F ) HIV-1 integration assessed by Alu-gag PCR (n=3). ( G ) In vivo detection of HIV and SIV DNA in human and non-human primate testicular germ cells by DNAscope. Representative pictures of TGC (green) harbouring viral DNA (red) within testis tissue section from one HIV-1 infected man (VL=51 681 cp/mL), one chronically infected rhesus macaque (VL > 10^6 cp/mL) and one African Green monkey 64 days post infection (VL=12085 cp/mL). Scale bars= 100μm.

Techniques Used: In Vivo, Infection, Selection, FACS, Negative Control, Purification, Incubation, Expressing, Polymerase Chain Reaction, Cell Culture

Characterisation and purity of primary TGCs ( A, B ) DRAQ5 DNA intercalating agent detection in flow cytometry on isolated TGCs showed the presence of haploid (n DNA, green), diploid (2n DNA, pink) and tetraploid cells (4n DNA, blue) (representative profile in A). ( C, E ) As expected, DDX4 labelled most germ cells irrespective of their ploidy, whereas MAGEA4 labelled early diploid and tetraploid germ cells (representative profile in C). ( D, E ) Somatic cell contaminants analysis in isolated TGC populations labeled with DRAQ-5 using flow cytometry and antibodies specific for somatic cells (class I HLA and vimentin) and leukocyte antigen CD45 (representative profile in D).
Figure Legend Snippet: Characterisation and purity of primary TGCs ( A, B ) DRAQ5 DNA intercalating agent detection in flow cytometry on isolated TGCs showed the presence of haploid (n DNA, green), diploid (2n DNA, pink) and tetraploid cells (4n DNA, blue) (representative profile in A). ( C, E ) As expected, DDX4 labelled most germ cells irrespective of their ploidy, whereas MAGEA4 labelled early diploid and tetraploid germ cells (representative profile in C). ( D, E ) Somatic cell contaminants analysis in isolated TGC populations labeled with DRAQ-5 using flow cytometry and antibodies specific for somatic cells (class I HLA and vimentin) and leukocyte antigen CD45 (representative profile in D).

Techniques Used: Flow Cytometry, Isolation, Labeling

20) Product Images from "Inflammatory Dendritic Cells Contribute to Regulate the Immune Response in Sickle Cell Disease"

Article Title: Inflammatory Dendritic Cells Contribute to Regulate the Immune Response in Sickle Cell Disease

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2020.617962

Activation profile and inflammatory cytokine production by mo-derived DCs from patients with SCD. Monocytes were isolated from the PBMCs of the patients (SS) and controls (AA) and cultured with 20 ng/ml GM-CSF and IL-4 for 6 to 7 days for DC differentiation. (A) Flow cytometry analysis of DC activation marker expression. (B) LEGENDplex measurement and flow cytometry acquisition of cytokine secretion in the supernatant after 24 h. (C) CD14 expression in CD209 + HLA-DR + DCs after differentiation. (D) Analysis of DC activation markers in the CD14 + and CD14 − DC subpopulations. P -values were obtained using the Mann-Whitney test. (E) CD14 + and CD14 − DCs from the same patient were sorted by flow cytometry and incubated for 24 h. Cytokines were measured in the supernatant by LEGENDplex. P -values were obtained using the paired Student’s t -test.
Figure Legend Snippet: Activation profile and inflammatory cytokine production by mo-derived DCs from patients with SCD. Monocytes were isolated from the PBMCs of the patients (SS) and controls (AA) and cultured with 20 ng/ml GM-CSF and IL-4 for 6 to 7 days for DC differentiation. (A) Flow cytometry analysis of DC activation marker expression. (B) LEGENDplex measurement and flow cytometry acquisition of cytokine secretion in the supernatant after 24 h. (C) CD14 expression in CD209 + HLA-DR + DCs after differentiation. (D) Analysis of DC activation markers in the CD14 + and CD14 − DC subpopulations. P -values were obtained using the Mann-Whitney test. (E) CD14 + and CD14 − DCs from the same patient were sorted by flow cytometry and incubated for 24 h. Cytokines were measured in the supernatant by LEGENDplex. P -values were obtained using the paired Student’s t -test.

Techniques Used: Activation Assay, Derivative Assay, Isolation, Cell Culture, Flow Cytometry, Marker, Expressing, MANN-WHITNEY, Incubation

Patients with SCD have an increased number of total DCs in the bloodstream and distinct proportions of DC subsets. The different DC subsets were identified in the blood from the patients (SS: hemoglobin S) and controls (AA: hemoglobin A) by flow cytometry. (A) The gating strategy used to identify total DCs, namely, lineage − (CD3/CD19/CD56/CD14) HLA-DR + ; pre-DCs (lineage − HLA-DR + CD1c − CD141 − CD123 − CD135 + ); cDC1 (lineage − HLA-DR + CD1c − CD141 + CD123 − ); cDC2 (lineage − HLA-DR + CD1c + CD141 − CD123 − ); pDC (lineage − HLA-DR + CD1c − CD141 − CD123 + ), and iDC is lineage − (CD3/CD19/CD56), HLA-DR + CD1c + CD14 + . (B) The percentage of total DCs in PBMCs and their absolute number per µl blood. (C) The percentage of pre-DCs in total DCs and their absolute number per µl blood. (D) The percentage within total DCs (top panel) and the absolute number per µl (bottom panel) cDC2, cDC1, pDC, and iDC. N: AA = 20; SS = 22. P -values were obtained using the Mann-Whitney test. (E) Spearman correlation between %iDC (left panel) and #iDC (right panel) with circulating reticulocyte numbers.
Figure Legend Snippet: Patients with SCD have an increased number of total DCs in the bloodstream and distinct proportions of DC subsets. The different DC subsets were identified in the blood from the patients (SS: hemoglobin S) and controls (AA: hemoglobin A) by flow cytometry. (A) The gating strategy used to identify total DCs, namely, lineage − (CD3/CD19/CD56/CD14) HLA-DR + ; pre-DCs (lineage − HLA-DR + CD1c − CD141 − CD123 − CD135 + ); cDC1 (lineage − HLA-DR + CD1c − CD141 + CD123 − ); cDC2 (lineage − HLA-DR + CD1c + CD141 − CD123 − ); pDC (lineage − HLA-DR + CD1c − CD141 − CD123 + ), and iDC is lineage − (CD3/CD19/CD56), HLA-DR + CD1c + CD14 + . (B) The percentage of total DCs in PBMCs and their absolute number per µl blood. (C) The percentage of pre-DCs in total DCs and their absolute number per µl blood. (D) The percentage within total DCs (top panel) and the absolute number per µl (bottom panel) cDC2, cDC1, pDC, and iDC. N: AA = 20; SS = 22. P -values were obtained using the Mann-Whitney test. (E) Spearman correlation between %iDC (left panel) and #iDC (right panel) with circulating reticulocyte numbers.

Techniques Used: Flow Cytometry, MANN-WHITNEY

21) Product Images from "Identification of immunodominant T cell epitopes of human glutamic acid decarboxylase 65 by using HLA-DR(?1*0101,?1*0401) transgenic mice"

Article Title: Identification of immunodominant T cell epitopes of human glutamic acid decarboxylase 65 by using HLA-DR(?1*0101,?1*0401) transgenic mice

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi:

The repertoire of mouse T cells in DR(α*0101,β*0401) transgenic mice. FACS analyses of peripheral blood lymphocytes from HLA-transgenic mice of different genetic background. Cells were stained using mouse CD4, hCD4, and human DR-specific reagents in standard protocols. FITC, fluorescein isothiocyanate; PE, phycoerythrin. ( Upper Left ) CD4 + T cell levels of progeny of DBA/1J mice backcrossed once with I-Aβ 0/0 mice (27% CD4 + T cells). ( Upper Right ) Analysis for a DR0401 mouse without hCD4 on an I-Aβ 0/0 background (3.8% CD4 + T cells). ( Lower Left and Right ) Heterozygous (13.3% CD4 + T cells) and homozygous (31.1% CD4 + T cells) HLA-DR, hCD4 mice, respectively. The DR4 and hCD4 are part of a single transgene in the C-line mice.
Figure Legend Snippet: The repertoire of mouse T cells in DR(α*0101,β*0401) transgenic mice. FACS analyses of peripheral blood lymphocytes from HLA-transgenic mice of different genetic background. Cells were stained using mouse CD4, hCD4, and human DR-specific reagents in standard protocols. FITC, fluorescein isothiocyanate; PE, phycoerythrin. ( Upper Left ) CD4 + T cell levels of progeny of DBA/1J mice backcrossed once with I-Aβ 0/0 mice (27% CD4 + T cells). ( Upper Right ) Analysis for a DR0401 mouse without hCD4 on an I-Aβ 0/0 background (3.8% CD4 + T cells). ( Lower Left and Right ) Heterozygous (13.3% CD4 + T cells) and homozygous (31.1% CD4 + T cells) HLA-DR, hCD4 mice, respectively. The DR4 and hCD4 are part of a single transgene in the C-line mice.

Techniques Used: Transgenic Assay, Mouse Assay, FACS, Staining

22) Product Images from "Vertebral body versus iliac crest bone marrow as a source of multipotential stromal cells: Comparison of processing techniques, tri-lineage differentiation and application on a scaffold for spine fusion"

Article Title: Vertebral body versus iliac crest bone marrow as a source of multipotential stromal cells: Comparison of processing techniques, tri-lineage differentiation and application on a scaffold for spine fusion

Journal: PLoS ONE

doi: 10.1371/journal.pone.0197969

Abundance, proliferation and phenotype of VB-BM versus IC-BM MSCs. A. The numbers of CD45 low CD271 high cells in unprocessed IC-BM and VB-BM aspirates (means are shown, Paired t-test, n = 6). B. The population doubling time (PDT) for VB-BM versus IC-BM MSCs (means are shown, Paired t-test, n = 7). C. The percentage of culture-expanded VB-BM MSCs versus IC-BM MSCs expressing hematopoietic lineage markers (CD34, CD14, CD19, HLA-DR), CD45, CD73, CD90, and CD105 (n = 3). The means of percentages are shown with bars of the standard error of the mean. D. The mean fluorescence intensity of positive markers expressed on VB-BM and IC-BM MSCs (n = 3).
Figure Legend Snippet: Abundance, proliferation and phenotype of VB-BM versus IC-BM MSCs. A. The numbers of CD45 low CD271 high cells in unprocessed IC-BM and VB-BM aspirates (means are shown, Paired t-test, n = 6). B. The population doubling time (PDT) for VB-BM versus IC-BM MSCs (means are shown, Paired t-test, n = 7). C. The percentage of culture-expanded VB-BM MSCs versus IC-BM MSCs expressing hematopoietic lineage markers (CD34, CD14, CD19, HLA-DR), CD45, CD73, CD90, and CD105 (n = 3). The means of percentages are shown with bars of the standard error of the mean. D. The mean fluorescence intensity of positive markers expressed on VB-BM and IC-BM MSCs (n = 3).

Techniques Used: Expressing, Fluorescence

23) Product Images from "Isolation of dental pulp stem cells from a single donor and characterization of their ability to differentiate after 2 years of cryopreservation"

Article Title: Isolation of dental pulp stem cells from a single donor and characterization of their ability to differentiate after 2 years of cryopreservation

Journal: Saudi Medical Journal

doi: 10.15537/smj.2016.5.13615

Isolated dental pulp stem cells characterization (DPSCs) showing A) Positive mesenchymal stem markers (CD105, CD90, CD73, CD13, CD29 and CD44) and negative for hematopoietic and endothelial markers (CD34, CD45, CD14, and CD31) as well as for the MHC class II marker human leukocyte antigen - antigen d related. B) Positive immunostaining for 4’, 6-diamidino-2-phenylindole (DAPI) and localization of Vimentin in DPSCs. C) Positive immunostaining for DAPI and negative for localization of Vimentin in control.
Figure Legend Snippet: Isolated dental pulp stem cells characterization (DPSCs) showing A) Positive mesenchymal stem markers (CD105, CD90, CD73, CD13, CD29 and CD44) and negative for hematopoietic and endothelial markers (CD34, CD45, CD14, and CD31) as well as for the MHC class II marker human leukocyte antigen - antigen d related. B) Positive immunostaining for 4’, 6-diamidino-2-phenylindole (DAPI) and localization of Vimentin in DPSCs. C) Positive immunostaining for DAPI and negative for localization of Vimentin in control.

Techniques Used: Isolation, Marker, Immunostaining

24) Product Images from "Monocytes and macrophages, targets of SARS-CoV-2: the clue for Covid-19 immunoparalysis"

Article Title: Monocytes and macrophages, targets of SARS-CoV-2: the clue for Covid-19 immunoparalysis

Journal: bioRxiv

doi: 10.1101/2020.09.17.300996

Peripheral blood mononuclear cells from Covid-19 patients were isolated and monocyte sub-populations were investigated by flow cytometry. ( A ) Non-classical, classical and intermediate HLA-DR + monocytes were evaluated from moderate, mild and severe Covid-19 clinical population. ( B ) Mean fluorescence intensity (MFI) of HLA-DR and CD163 expression was investigated for CD14 + , CD14 + /CD16 + and CD16 + monocyte populations from moderate, mild and severe Covid-19 patients using t-test.
Figure Legend Snippet: Peripheral blood mononuclear cells from Covid-19 patients were isolated and monocyte sub-populations were investigated by flow cytometry. ( A ) Non-classical, classical and intermediate HLA-DR + monocytes were evaluated from moderate, mild and severe Covid-19 clinical population. ( B ) Mean fluorescence intensity (MFI) of HLA-DR and CD163 expression was investigated for CD14 + , CD14 + /CD16 + and CD16 + monocyte populations from moderate, mild and severe Covid-19 patients using t-test.

Techniques Used: Isolation, Flow Cytometry, Fluorescence, Expressing

Monocyte subsets are altered in SARS-CoV-2-infected patients. PBMCs from healthy donors and Covid-19 patients were isolated and monocyte sub-populations were investigated by flow cytometry ( A ) Representative flow cytometry plot showing the gating strategy to investigate non-classical, classical and intermediate HLA-DR + monocytes from Covid-19 patients and healthy donors as control. ( B ) Mean fluorescence intensity (MFI) of HLA-DR and CD163 expression was investigated for CD14 + , CD14 + /CD16 + and CD16 + monocyte populations from healthy and Covid-19 patients. ( C ) Monocytes from healthy donors were stimulated with SARS-CoV-2 IHU-MI3 strain (0.25 or 0.5 MOI). The expression of HLA-DR and CD163 was observed at 24 and 48 hours of infection. ** P
Figure Legend Snippet: Monocyte subsets are altered in SARS-CoV-2-infected patients. PBMCs from healthy donors and Covid-19 patients were isolated and monocyte sub-populations were investigated by flow cytometry ( A ) Representative flow cytometry plot showing the gating strategy to investigate non-classical, classical and intermediate HLA-DR + monocytes from Covid-19 patients and healthy donors as control. ( B ) Mean fluorescence intensity (MFI) of HLA-DR and CD163 expression was investigated for CD14 + , CD14 + /CD16 + and CD16 + monocyte populations from healthy and Covid-19 patients. ( C ) Monocytes from healthy donors were stimulated with SARS-CoV-2 IHU-MI3 strain (0.25 or 0.5 MOI). The expression of HLA-DR and CD163 was observed at 24 and 48 hours of infection. ** P

Techniques Used: Infection, Isolation, Flow Cytometry, Fluorescence, Expressing

25) Product Images from "Preferential Recruitment of Th17 Cells to Cervical Cancer via CCR6-CCL20 Pathway"

Article Title: Preferential Recruitment of Th17 Cells to Cervical Cancer via CCR6-CCL20 Pathway

Journal: PLoS ONE

doi: 10.1371/journal.pone.0120855

Phenotypic analysis of Th17 cells in patients with cervical cancer. (A) Representative expression profiles of CD45RO, HLA-DR, Granzyme B and PD-1 in tumor-infiltrating Th17 cells. The percentages represent the frequencies of various markers in Th17 cells. (B) Representative expression profiles of CCR4, CCR6 and CD49d on Th17 cells from peripheral blood (long dotted line), non-tumor (dotted line) and tumor tissues (solid line). The percentages represent the frequencies of various markers on tumor-infiltrating Th17 cells. (C) Statistical analysis of surface expression of CCR4, CCR6, CD49d on Th17 cells from peripheral blood, non-tumor and tumor tissues (n = 25). * P
Figure Legend Snippet: Phenotypic analysis of Th17 cells in patients with cervical cancer. (A) Representative expression profiles of CD45RO, HLA-DR, Granzyme B and PD-1 in tumor-infiltrating Th17 cells. The percentages represent the frequencies of various markers in Th17 cells. (B) Representative expression profiles of CCR4, CCR6 and CD49d on Th17 cells from peripheral blood (long dotted line), non-tumor (dotted line) and tumor tissues (solid line). The percentages represent the frequencies of various markers on tumor-infiltrating Th17 cells. (C) Statistical analysis of surface expression of CCR4, CCR6, CD49d on Th17 cells from peripheral blood, non-tumor and tumor tissues (n = 25). * P

Techniques Used: Expressing

26) Product Images from "Macrophages in Vaginal but Not Intestinal Mucosa Are Monocyte-Like and Permissive to Human Immunodeficiency Virus Type 1 Infection ▿"

Article Title: Macrophages in Vaginal but Not Intestinal Mucosa Are Monocyte-Like and Permissive to Human Immunodeficiency Virus Type 1 Infection ▿

Journal: Journal of Virology

doi: 10.1128/JVI.01796-08

Vaginal, but not intestinal, macrophages retain a monocyte-like phenotype. Gradient sedimentation purified blood and mucosal MNLs were stained with fluorescence-conjugated antibodies to the myeloid markers HLA-DR and CD13 and the innate receptors CD14,
Figure Legend Snippet: Vaginal, but not intestinal, macrophages retain a monocyte-like phenotype. Gradient sedimentation purified blood and mucosal MNLs were stained with fluorescence-conjugated antibodies to the myeloid markers HLA-DR and CD13 and the innate receptors CD14,

Techniques Used: Sedimentation, Purification, Staining, Fluorescence

27) Product Images from "Characterization of menstrual stem cells: angiogenic effect, migration and hematopoietic stem cell support in comparison with bone marrow mesenchymal stem cells"

Article Title: Characterization of menstrual stem cells: angiogenic effect, migration and hematopoietic stem cell support in comparison with bone marrow mesenchymal stem cells

Journal: Stem Cell Research & Therapy

doi: 10.1186/s13287-015-0013-5

MenSCs show high expression level of mesodermal antigens and multilineage capacities. (A) MenSCs display stem cell-like phenotypic markers. In order to determine the immunophenotype MenSCs and BM-MSCs were stained by conjugated antibodies against mesenchymal, hematopoietic and pluripotential stem cells markers, then analyzed by FACS. MenSCs (orange filled histogram) and BM-MSCs (green filled histogram) displayed positive expression for mesenchymal stromal cells while being negative for other markers, such as CD14, CD34, CD45. Besides, low expression for CD146 and high expression for HLA-ABC and CD49a were observed in MenSCs samples. In addition, FACS analysis showed that both cells do not express pluripotent surface markers, such as SSEA-3, SSEA-4, and TRA-1-60. Isotype-matched controls of MenSCs and BM-MSCs are shown with a blue and red filled histogram, respectively. B) Differentiation potential of BM-MSCs and MenSCs. Adipose differentiation was characterized according to the fold increased level of the peroxysome proliferator-activated receptor (PPAR)-γ and by the formation of lipid droplets that are positive for Oil red O staining. For osteogenesis characterization, the fold increase level of osteocalcin (OC) and positive staining for alizarin red were evaluated. Chondrogenesis was tested by the expression of collagen IIA (col 2A) and aggrecan (Agg) and by positive staining for Safranin O. Values are expressed as mean ± SE of triplicates (* P ≤0.05). Data shown are representative of multiple replicates. BM-MSCs, bone marrow-derived mesenchymal stem cells; FACS, fluorescence-activated cell sorting; MenSCs, menstrual-drived stem cells; SE, standard error.
Figure Legend Snippet: MenSCs show high expression level of mesodermal antigens and multilineage capacities. (A) MenSCs display stem cell-like phenotypic markers. In order to determine the immunophenotype MenSCs and BM-MSCs were stained by conjugated antibodies against mesenchymal, hematopoietic and pluripotential stem cells markers, then analyzed by FACS. MenSCs (orange filled histogram) and BM-MSCs (green filled histogram) displayed positive expression for mesenchymal stromal cells while being negative for other markers, such as CD14, CD34, CD45. Besides, low expression for CD146 and high expression for HLA-ABC and CD49a were observed in MenSCs samples. In addition, FACS analysis showed that both cells do not express pluripotent surface markers, such as SSEA-3, SSEA-4, and TRA-1-60. Isotype-matched controls of MenSCs and BM-MSCs are shown with a blue and red filled histogram, respectively. B) Differentiation potential of BM-MSCs and MenSCs. Adipose differentiation was characterized according to the fold increased level of the peroxysome proliferator-activated receptor (PPAR)-γ and by the formation of lipid droplets that are positive for Oil red O staining. For osteogenesis characterization, the fold increase level of osteocalcin (OC) and positive staining for alizarin red were evaluated. Chondrogenesis was tested by the expression of collagen IIA (col 2A) and aggrecan (Agg) and by positive staining for Safranin O. Values are expressed as mean ± SE of triplicates (* P ≤0.05). Data shown are representative of multiple replicates. BM-MSCs, bone marrow-derived mesenchymal stem cells; FACS, fluorescence-activated cell sorting; MenSCs, menstrual-drived stem cells; SE, standard error.

Techniques Used: Expressing, Staining, FACS, Derivative Assay, Fluorescence

28) Product Images from "Chimeric Antigen Receptor T Cells Targeting NKG2D-Ligands Show Robust Efficacy Against Acute Myeloid Leukemia and T-Cell Acute Lymphoblastic Leukemia"

Article Title: Chimeric Antigen Receptor T Cells Targeting NKG2D-Ligands Show Robust Efficacy Against Acute Myeloid Leukemia and T-Cell Acute Lymphoblastic Leukemia

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2020.580328

NKG2D-ligands are frequently expressed in AML and T-ALL (A) The NKG2D ligand surface expression in primary AML bone marrow aspirate samples and AML cell lines (MV4-11, Molm-13, HL-60) was analyzed by flow cytometry using the conjugated fusion protein NKG2D-Fc (red) and IgG-Fc isotype (blue) to evaluate ligand detection. The K562 cell line was included as a positive control due to its high level of NKG2D-ligand expression. Primary AML blasts were identified based on viability, CD45dim expression and known patient specific markers including CD117, CD34, HLA-DR, and CD33. Histograms illustrate NKG2D-Fc (red) vs. IgG-Fc (blue) isotype control staining in the respective samples (B) Histograms show NKG2D-Fc (red) vs. IgG-Fc (blue) isotype control staining in primary T-All blasts and T-ALL cell lines (KOPT-K1, DND-41, HPB-ALL, Jurkat). Primary T-ALL blasts were identified based on viability and a combination of CD45, CD2, CD3, CD5, CD7, CD34 and CD8 according to their patient-specific clinical expression pattern (C) Histogram shows NKG2D-Fc (red) vs. IgG-Fc (blue) isotype control staining in the murine melanoma cell line B16 (D) The intensity of NKG2D-Ligand expression is shown as specific fluorescence intensity (SFI), which represents the ratio of the mean fluorescence intensity (MFI) of NKG2D-Fc to IgG-Fc. All examined primary samples (AML n=24) and one representative sample for each of the cell lines (n ≥ 3 for each cell line) are displayed in comparison. SFI > 1 is indicative of NKG2D-ligand expression above isotype (E) Staining of primary AML samples with available antibodies to the individual NKG2D-ligands was also undertaken and is displayed here in comparison (F) SFIs of all examined primary T-ALL samples (n=6) and one representative sample for each of the cell lines (n ≥ 3 for each cell line) are shown (G) Individual NKG2D-ligand staining with available antibodies was also performed on the cell lines indicated.
Figure Legend Snippet: NKG2D-ligands are frequently expressed in AML and T-ALL (A) The NKG2D ligand surface expression in primary AML bone marrow aspirate samples and AML cell lines (MV4-11, Molm-13, HL-60) was analyzed by flow cytometry using the conjugated fusion protein NKG2D-Fc (red) and IgG-Fc isotype (blue) to evaluate ligand detection. The K562 cell line was included as a positive control due to its high level of NKG2D-ligand expression. Primary AML blasts were identified based on viability, CD45dim expression and known patient specific markers including CD117, CD34, HLA-DR, and CD33. Histograms illustrate NKG2D-Fc (red) vs. IgG-Fc (blue) isotype control staining in the respective samples (B) Histograms show NKG2D-Fc (red) vs. IgG-Fc (blue) isotype control staining in primary T-All blasts and T-ALL cell lines (KOPT-K1, DND-41, HPB-ALL, Jurkat). Primary T-ALL blasts were identified based on viability and a combination of CD45, CD2, CD3, CD5, CD7, CD34 and CD8 according to their patient-specific clinical expression pattern (C) Histogram shows NKG2D-Fc (red) vs. IgG-Fc (blue) isotype control staining in the murine melanoma cell line B16 (D) The intensity of NKG2D-Ligand expression is shown as specific fluorescence intensity (SFI), which represents the ratio of the mean fluorescence intensity (MFI) of NKG2D-Fc to IgG-Fc. All examined primary samples (AML n=24) and one representative sample for each of the cell lines (n ≥ 3 for each cell line) are displayed in comparison. SFI > 1 is indicative of NKG2D-ligand expression above isotype (E) Staining of primary AML samples with available antibodies to the individual NKG2D-ligands was also undertaken and is displayed here in comparison (F) SFIs of all examined primary T-ALL samples (n=6) and one representative sample for each of the cell lines (n ≥ 3 for each cell line) are shown (G) Individual NKG2D-ligand staining with available antibodies was also performed on the cell lines indicated.

Techniques Used: Expressing, Flow Cytometry, Positive Control, Staining, Fluorescence

29) Product Images from "Coexpression of CD163 and CD141 identifies human circulating IL-10-producing dendritic cells (DC-10)"

Article Title: Coexpression of CD163 and CD141 identifies human circulating IL-10-producing dendritic cells (DC-10)

Journal: Cellular and Molecular Immunology

doi: 10.1038/s41423-019-0218-0

Ex vivo DC-10 promote hypo-responsiveness in allogeneic naïve CD4 + T cells. a Scheme of the experiment: naïve CD4 + T cells were cultured with ex vivo DC-10 (CD14 + CD16 + CD141 + CD163 + ) or ex vivo cDCs (CD1c + CD11c + ) FACS isolated from the peripheral blood of healthy donors (ratio 10:1) for 5 days. b Expression of the activation markers CD25 and HLA-DR on CD4 + T cells stimulated with ex vivo cDCs (red) or ex vivo DC-10 (blue) were evaluated by flow cytometry ( n = 6). The following gating strategy was applied: doublet exclusion, followed by alive cells and gating on CD3 + CD4 + cells. Left, each dot represents a single donor; right, flow cytometry histograms from one representative donor. c Naive CD4 + T cell proliferation was evaluated by proliferation dye dilution ( n = 8). The following gating strategy was applied: doublet exclusion, followed by alive cells and gating on CD3 + CD4 + cells. The percentage of proliferated cells is shown for each single donor (left panel) and in representative dot plots (right panels). d IFN-γ and GM-CSF in culture supernatants were measured by ELISA ( n = 8). Each dot represents a single donor. * P ≤ 0.05 ** P ≤ 0.01 (Wilcoxon matched pairs test, two-tailed)
Figure Legend Snippet: Ex vivo DC-10 promote hypo-responsiveness in allogeneic naïve CD4 + T cells. a Scheme of the experiment: naïve CD4 + T cells were cultured with ex vivo DC-10 (CD14 + CD16 + CD141 + CD163 + ) or ex vivo cDCs (CD1c + CD11c + ) FACS isolated from the peripheral blood of healthy donors (ratio 10:1) for 5 days. b Expression of the activation markers CD25 and HLA-DR on CD4 + T cells stimulated with ex vivo cDCs (red) or ex vivo DC-10 (blue) were evaluated by flow cytometry ( n = 6). The following gating strategy was applied: doublet exclusion, followed by alive cells and gating on CD3 + CD4 + cells. Left, each dot represents a single donor; right, flow cytometry histograms from one representative donor. c Naive CD4 + T cell proliferation was evaluated by proliferation dye dilution ( n = 8). The following gating strategy was applied: doublet exclusion, followed by alive cells and gating on CD3 + CD4 + cells. The percentage of proliferated cells is shown for each single donor (left panel) and in representative dot plots (right panels). d IFN-γ and GM-CSF in culture supernatants were measured by ELISA ( n = 8). Each dot represents a single donor. * P ≤ 0.05 ** P ≤ 0.01 (Wilcoxon matched pairs test, two-tailed)

Techniques Used: Ex Vivo, Cell Culture, FACS, Isolation, Expressing, Activation Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test

30) Product Images from "Combinatorial Cytokine Code Generates Anti-Viral State in Dendritic Cells"

Article Title: Combinatorial Cytokine Code Generates Anti-Viral State in Dendritic Cells

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2014.00073

Effects of combinatorial of IFNβ, TNFα, and IL1β treatment on DCs infected with the pandemic influenza A Cal/09 strain . (A) Infectivity assayed by NP expression (* p ≤ 0.05 compared to cells exposed to IFNβ). (B) Expression of the maturation marker CD86. (C) Expression of the maturation marker HLA-ABC. (D) Expression of the maturation marker HLA-DR (* p ≤ 0.05 compared to cells exposed to the triple combination of TNFα, IFNβ, and IL1b). (E) Apoptosis assayed by assessing nuclear fragmentation and cell granularity by imaging flow cytometry (* p ≤ 0.05 compared to untreated cells). (F) Sample images of non-apoptotic cells. (G) Sample images of apoptotic cells.
Figure Legend Snippet: Effects of combinatorial of IFNβ, TNFα, and IL1β treatment on DCs infected with the pandemic influenza A Cal/09 strain . (A) Infectivity assayed by NP expression (* p ≤ 0.05 compared to cells exposed to IFNβ). (B) Expression of the maturation marker CD86. (C) Expression of the maturation marker HLA-ABC. (D) Expression of the maturation marker HLA-DR (* p ≤ 0.05 compared to cells exposed to the triple combination of TNFα, IFNβ, and IL1b). (E) Apoptosis assayed by assessing nuclear fragmentation and cell granularity by imaging flow cytometry (* p ≤ 0.05 compared to untreated cells). (F) Sample images of non-apoptotic cells. (G) Sample images of apoptotic cells.

Techniques Used: Infection, Expressing, Marker, Imaging, Flow Cytometry, Cytometry

Effects of combinatorial cytokine treatment . (A) Maturation marker expression after exposure to combinations of IFNβ, IL28, TNFSF15, TNFα and IL1β, measured by flow cytometry (* p ≤ 0.05 compared to untreated cells). (B) CD86 expression after exposure to combinations of IFNβ, TNFSF15, TNFα and IL1β, measured by flow cytometry (* p ≤ 0.05 compared to cells treated with IFNβ alone). (C) HLA-DR expression after exposure to combinations of IFNβ, TNFSF15, TNFα and IL1β, measured by flow cytometry (* p ≤ 0.05 compared to cells treated with IFNβ alone). (D) Infectivity of NDV after cytokine combination pretreatment for 8 h in DCs measured by RFP expression. (E) Phagocytosis assay measured by bead uptake by imaging flow cytometry (* p ≤ 0.05 compared to untreated cells).
Figure Legend Snippet: Effects of combinatorial cytokine treatment . (A) Maturation marker expression after exposure to combinations of IFNβ, IL28, TNFSF15, TNFα and IL1β, measured by flow cytometry (* p ≤ 0.05 compared to untreated cells). (B) CD86 expression after exposure to combinations of IFNβ, TNFSF15, TNFα and IL1β, measured by flow cytometry (* p ≤ 0.05 compared to cells treated with IFNβ alone). (C) HLA-DR expression after exposure to combinations of IFNβ, TNFSF15, TNFα and IL1β, measured by flow cytometry (* p ≤ 0.05 compared to cells treated with IFNβ alone). (D) Infectivity of NDV after cytokine combination pretreatment for 8 h in DCs measured by RFP expression. (E) Phagocytosis assay measured by bead uptake by imaging flow cytometry (* p ≤ 0.05 compared to untreated cells).

Techniques Used: Marker, Expressing, Flow Cytometry, Cytometry, Infection, Phagocytosis Assay, Imaging

31) Product Images from "Activated monocytes in peritumoral stroma of hepatocellular carcinoma foster immune privilege and disease progression through PD-L1"

Article Title: Activated monocytes in peritumoral stroma of hepatocellular carcinoma foster immune privilege and disease progression through PD-L1

Journal: The Journal of Experimental Medicine

doi: 10.1084/jem.20082173

Expression pattern of PD-L1 was correlated with the activation pattern of monocytes/Mφ in peritumoral stroma. (A–C) FACS analysis of PD-L1 and HLA-DR expression in fresh monocytes isolated from peripheral blood and tissues. Representative data on PD-L1 + HLA-DR high monocytes (A), percentages of HLA-DR high CD14 high monocytes (B), and the MFI of HLA-DR in HLA-DR high CD14 high monocytes (C) are shown. The samples collected and the numbers of donors in A–C were the same as in Fig. 1 . The data shown in A are representative dot plots of at least seven individuals from more than five independent experiments; B and C show the statistics analysis of these samples. The continuous and dashed horizontal bars in B represent median values. Results are expressed as means ± SEM. (D) Positive correlations between the levels of PD-L1 + and HLA-DR high monocytes. The samples used in D were blood from healthy individuals, HCC patients, and paired nontumor and tumor tissues from HCC patients ( n = 28 for each). (E) Adjacent sections of paraffin-embedded hepatoma samples stained with the indicated markers. The micrographs at higher magnification show the peritumoral stroma region (1) and a cancer nest (2). Significant difference compared with healthy blood is indicated (***, P
Figure Legend Snippet: Expression pattern of PD-L1 was correlated with the activation pattern of monocytes/Mφ in peritumoral stroma. (A–C) FACS analysis of PD-L1 and HLA-DR expression in fresh monocytes isolated from peripheral blood and tissues. Representative data on PD-L1 + HLA-DR high monocytes (A), percentages of HLA-DR high CD14 high monocytes (B), and the MFI of HLA-DR in HLA-DR high CD14 high monocytes (C) are shown. The samples collected and the numbers of donors in A–C were the same as in Fig. 1 . The data shown in A are representative dot plots of at least seven individuals from more than five independent experiments; B and C show the statistics analysis of these samples. The continuous and dashed horizontal bars in B represent median values. Results are expressed as means ± SEM. (D) Positive correlations between the levels of PD-L1 + and HLA-DR high monocytes. The samples used in D were blood from healthy individuals, HCC patients, and paired nontumor and tumor tissues from HCC patients ( n = 28 for each). (E) Adjacent sections of paraffin-embedded hepatoma samples stained with the indicated markers. The micrographs at higher magnification show the peritumoral stroma region (1) and a cancer nest (2). Significant difference compared with healthy blood is indicated (***, P

Techniques Used: Expressing, Activation Assay, FACS, Isolation, Staining

32) Product Images from "Differential plasmacytoid dendritic cell phenotype and type I Interferon response in asymptomatic and severe COVID-19 infection"

Article Title: Differential plasmacytoid dendritic cell phenotype and type I Interferon response in asymptomatic and severe COVID-19 infection

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1009878

pDC differently activate and express chemokine receptors in COVID-19 asymptomatic and hospitalized patients. Freshly isolated peripheral blood mononuclear cells (PBMC) from asymptomatic (CP-AS, n = 8) and hospitalized COVID-19 patients (CP, n = 6) as well as matched healthy donors (HD, n = 5) were stained with a cocktails of antibodies to study by flow cytometry plasmacytoid dendritic cell (pDC) frequency and absolute number ( A ), diversification and activation status (Lineage, CD123, BDCA-4, HLA-DR, PD-L1, CD80 and CD86) ( B, C, D ), as well as expression of chemokine receptors (CXCR4, CXCR3, CCR7 and CD62-L) ( E, F ). The percentage (%) of shown pDC sub-populations ( B, C, E ) was evaluated in live/single Lineage - CD123 + BDCA4 + HLADR + gated pDC and depicted for each studied patients or HD together with mean ± SEM values. Surface expression of PD-L1, CD80, CD86 ( D ) and CXCR4, CXCR3, CD62-L, CCR7 ( F ), was determined as mean fluorescence intensity (MFI) and shown results were mean relative values ± SEM of analyzed patients or HD. P-values were depicted as follows: *p≤0.05; ** p≤0.001; *** p≤0.0001.
Figure Legend Snippet: pDC differently activate and express chemokine receptors in COVID-19 asymptomatic and hospitalized patients. Freshly isolated peripheral blood mononuclear cells (PBMC) from asymptomatic (CP-AS, n = 8) and hospitalized COVID-19 patients (CP, n = 6) as well as matched healthy donors (HD, n = 5) were stained with a cocktails of antibodies to study by flow cytometry plasmacytoid dendritic cell (pDC) frequency and absolute number ( A ), diversification and activation status (Lineage, CD123, BDCA-4, HLA-DR, PD-L1, CD80 and CD86) ( B, C, D ), as well as expression of chemokine receptors (CXCR4, CXCR3, CCR7 and CD62-L) ( E, F ). The percentage (%) of shown pDC sub-populations ( B, C, E ) was evaluated in live/single Lineage - CD123 + BDCA4 + HLADR + gated pDC and depicted for each studied patients or HD together with mean ± SEM values. Surface expression of PD-L1, CD80, CD86 ( D ) and CXCR4, CXCR3, CD62-L, CCR7 ( F ), was determined as mean fluorescence intensity (MFI) and shown results were mean relative values ± SEM of analyzed patients or HD. P-values were depicted as follows: *p≤0.05; ** p≤0.001; *** p≤0.0001.

Techniques Used: Isolation, Staining, Flow Cytometry, Activation Assay, Expressing, Fluorescence

33) Product Images from "Triggering receptor expressed on myeloid cells - 1 (Trem-1) on blood neutrophils is associated with cytokine inducibility in human E. coli sepsis"

Article Title: Triggering receptor expressed on myeloid cells - 1 (Trem-1) on blood neutrophils is associated with cytokine inducibility in human E. coli sepsis

Journal: Diagnostic Pathology

doi: 10.1186/1746-1596-8-24

A: Flow cytometry expression of TLR2, TLR4, CD14, HLA-DR, Trem-1 on monocytes. MFI ± SD is shown for patients (P) and healthy volunteers (V); *
Figure Legend Snippet: A: Flow cytometry expression of TLR2, TLR4, CD14, HLA-DR, Trem-1 on monocytes. MFI ± SD is shown for patients (P) and healthy volunteers (V); *

Techniques Used: Flow Cytometry, Cytometry, Expressing

34) Product Images from "Transcriptome analysis reveals similarities between human blood CD3− CD56bright cells and mouse CD127+ innate lymphoid cells"

Article Title: Transcriptome analysis reveals similarities between human blood CD3− CD56bright cells and mouse CD127+ innate lymphoid cells

Journal: Scientific Reports

doi: 10.1038/s41598-017-03256-0

Expression differences between mouse splenic CD127 + NK1.1 + NKp46 + ILC and CD127 − NK1.1 + NKp46 + NK cells show commonalities with differences between human blood CD56 bright and CD56 dim cells. ImmGen microarray data examining mouse splenic NK1.1 + NKp46 + CD127 + CD27 + cells (annotated as ILC1) and NK1.1 + NKp46 + CD127 − CD27 +/− NK cells 23 is analysed to show fold-change difference between subsets (x-axis). For all mouse genes with corresponding human genes in the NCBI Homologene database, this is contrasted with microarray expression differences observed between human blood CD56 bright and CD56 dim populations (y-axis) (HTA2.0 microarray) 21 . Transcripts differing by ≥2-fold between both human and mouse subsets are labelled, and their numbers are shown. Transcripts similarly varying by ≥2-fold in mouse ILC1/NK comparisons in both liver and small intestine (in addition to spleen) are shown in blue; those differing by ≥2-fold in 2-of-3 comparisons of mouse tissues are in green (see Suppl. Figure 3 ). Bold names also show ≥2-fold differences in an independent microarray dataset comparing human CD56 bright and CD56 dim cells (see Suppl. Figure 4 ). Although not matched by Homologene, human HLA-DRA (paired with mouse H2-Aa) is shown (with asterisk) but not included in transcript counts. ( B ) Flow cytometry staining of gated human blood CD56 bright CD16 − CD3 − or CD56 dim CD16 + CD3 − populations from three donors. Quadrant gates were set using fluorescence minus one (FMO) stains.
Figure Legend Snippet: Expression differences between mouse splenic CD127 + NK1.1 + NKp46 + ILC and CD127 − NK1.1 + NKp46 + NK cells show commonalities with differences between human blood CD56 bright and CD56 dim cells. ImmGen microarray data examining mouse splenic NK1.1 + NKp46 + CD127 + CD27 + cells (annotated as ILC1) and NK1.1 + NKp46 + CD127 − CD27 +/− NK cells 23 is analysed to show fold-change difference between subsets (x-axis). For all mouse genes with corresponding human genes in the NCBI Homologene database, this is contrasted with microarray expression differences observed between human blood CD56 bright and CD56 dim populations (y-axis) (HTA2.0 microarray) 21 . Transcripts differing by ≥2-fold between both human and mouse subsets are labelled, and their numbers are shown. Transcripts similarly varying by ≥2-fold in mouse ILC1/NK comparisons in both liver and small intestine (in addition to spleen) are shown in blue; those differing by ≥2-fold in 2-of-3 comparisons of mouse tissues are in green (see Suppl. Figure 3 ). Bold names also show ≥2-fold differences in an independent microarray dataset comparing human CD56 bright and CD56 dim cells (see Suppl. Figure 4 ). Although not matched by Homologene, human HLA-DRA (paired with mouse H2-Aa) is shown (with asterisk) but not included in transcript counts. ( B ) Flow cytometry staining of gated human blood CD56 bright CD16 − CD3 − or CD56 dim CD16 + CD3 − populations from three donors. Quadrant gates were set using fluorescence minus one (FMO) stains.

Techniques Used: Expressing, Microarray, Flow Cytometry, Cytometry, Staining, Fluorescence

35) Product Images from "Antigen-Presenting Cells Are Stratified Within Normal Human Corneas and Are Rapidly Mobilized During Ex Vivo Viral Infection"

Article Title: Antigen-Presenting Cells Are Stratified Within Normal Human Corneas and Are Rapidly Mobilized During Ex Vivo Viral Infection

Journal: Investigative Ophthalmology & Visual Science

doi: 10.1167/iovs.13-13523

Phenotype of APC within the normal human cornea. ( A , B ) Fresh donor human corneal tissue was dispersed with collagenase prior to flow cytometric analysis. ( A ) Dot plots showing gating strategy to identify CD45 + , CD11c + , and HLA-DR + cells. SSC, side scatter.
Figure Legend Snippet: Phenotype of APC within the normal human cornea. ( A , B ) Fresh donor human corneal tissue was dispersed with collagenase prior to flow cytometric analysis. ( A ) Dot plots showing gating strategy to identify CD45 + , CD11c + , and HLA-DR + cells. SSC, side scatter.

Techniques Used: Flow Cytometry

36) Product Images from "Oral Wild-Type Salmonella Typhi Challenge Induces Activation of Circulating Monocytes and Dendritic Cells in Individuals Who Develop Typhoid Disease"

Article Title: Oral Wild-Type Salmonella Typhi Challenge Induces Activation of Circulating Monocytes and Dendritic Cells in Individuals Who Develop Typhoid Disease

Journal: PLoS Neglected Tropical Diseases

doi: 10.1371/journal.pntd.0003837

Gating strategy. Stained PBMC were gated on cells with high FSC and SSC characteristics avoiding the lymphocyte region (A), doublets/multiplets were eliminated (B) and subsequently live cells were gated based on expression of CD14 (C). The viability channel (live/dead-L/D-) also contained CD3; therefore cells selected were also CD3 - (C). CD14+ cells also expressed CD16 and HLA-DR. Therefore, these cells were classified as monocytes (D). All CD3 - CD14 - expressed HLA-DR and a percentage of these cells expressed either CD11c or CD123 but lacked CD19, CD66b, CD16 and CD56; therefore, these cells were classified as dendritic cells (DC) (E) (see also S4 Fig ). The identity of these cells as DCs was confirmed using PBMC from healthy volunteers stained with monoclonal antibodies to BDCA-1, BDCA-2 and BDCA-3 ( S4 Fig ). Plots shown are from a representative volunteer. Gray histograms represent fluorescent minus one (FMO) samples.
Figure Legend Snippet: Gating strategy. Stained PBMC were gated on cells with high FSC and SSC characteristics avoiding the lymphocyte region (A), doublets/multiplets were eliminated (B) and subsequently live cells were gated based on expression of CD14 (C). The viability channel (live/dead-L/D-) also contained CD3; therefore cells selected were also CD3 - (C). CD14+ cells also expressed CD16 and HLA-DR. Therefore, these cells were classified as monocytes (D). All CD3 - CD14 - expressed HLA-DR and a percentage of these cells expressed either CD11c or CD123 but lacked CD19, CD66b, CD16 and CD56; therefore, these cells were classified as dendritic cells (DC) (E) (see also S4 Fig ). The identity of these cells as DCs was confirmed using PBMC from healthy volunteers stained with monoclonal antibodies to BDCA-1, BDCA-2 and BDCA-3 ( S4 Fig ). Plots shown are from a representative volunteer. Gray histograms represent fluorescent minus one (FMO) samples.

Techniques Used: Staining, Expressing

37) Product Images from "A Novel in vitro Human Macrophage Model to Study the Persistence of Mycobacterium tuberculosis Using Vitamin D3 and Retinoic Acid Activated THP-1 Macrophages"

Article Title: A Novel in vitro Human Macrophage Model to Study the Persistence of Mycobacterium tuberculosis Using Vitamin D3 and Retinoic Acid Activated THP-1 Macrophages

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2011.00067

Phorbol myristyl acetate (PMA) and a combination of retinoic acid (RA) and vitamin D 3 (VD) induce a differential expression of surface receptors on human THP-1 monocytoid cells . THPs were activated with varying doses (as indicated; C ) of PMA or a mix of RA and VD for 72 h and were stained for surface receptors using either fluorescent antibodies (A–C) or fluorescent tagged ligands (D,E) and analyzed using Cellquest software. MFIs from three experiments were calculated to show significance as indicated (*MFI significantly increased in three experiments; t test). (A) THPs were activated and stained for receptors involved in uptake or binding to mycobacteria [DC-SIGN, CD14, and CD44] (B) chemokine receptors, CD184 and CD195; and (C) CD80, CD86, CD1d, and HLA-DR receptors involved in antigen presentation. (D) THPs were activated and stained with mannosylated-BSA-FITC (mBSA-FITC) or BSA-FITC at 37°C for active uptake and at 4°C for absorption control using flow cytometry as above. RAVD enhanced the expression of macrophage mannose receptor compared to PMA activated THPs.
Figure Legend Snippet: Phorbol myristyl acetate (PMA) and a combination of retinoic acid (RA) and vitamin D 3 (VD) induce a differential expression of surface receptors on human THP-1 monocytoid cells . THPs were activated with varying doses (as indicated; C ) of PMA or a mix of RA and VD for 72 h and were stained for surface receptors using either fluorescent antibodies (A–C) or fluorescent tagged ligands (D,E) and analyzed using Cellquest software. MFIs from three experiments were calculated to show significance as indicated (*MFI significantly increased in three experiments; t test). (A) THPs were activated and stained for receptors involved in uptake or binding to mycobacteria [DC-SIGN, CD14, and CD44] (B) chemokine receptors, CD184 and CD195; and (C) CD80, CD86, CD1d, and HLA-DR receptors involved in antigen presentation. (D) THPs were activated and stained with mannosylated-BSA-FITC (mBSA-FITC) or BSA-FITC at 37°C for active uptake and at 4°C for absorption control using flow cytometry as above. RAVD enhanced the expression of macrophage mannose receptor compared to PMA activated THPs.

Techniques Used: Expressing, Staining, Software, Binding Assay, Flow Cytometry, Cytometry

38) Product Images from "Potential use of human adipose mesenchymal stromal cells for intervertebral disc regeneration: a preliminary study on biglycan-deficient murine model of chronic disc degeneration"

Article Title: Potential use of human adipose mesenchymal stromal cells for intervertebral disc regeneration: a preliminary study on biglycan-deficient murine model of chronic disc degeneration

Journal: Arthritis Research & Therapy

doi: 10.1186/s13075-014-0457-5

Features and immunophenotypic mesenchymal profile of human-derived adipose mesenchymal stem cell (ADSCs). (A) Immunophenotypic mesenchymal profile of ADSCs shows that these cells express a high percentage of positivity for CD73, CD90, CD105, and a low-positivity fraction for CD14, CD19, CD34, CD45 and HLA-DR. After selective stimulation, ADSCs exhibited adipogenic, chondrogenic and osteogenic potential, which was demonstrated by Oil Red O staining of lipid droplets (B) , aggrecan deposition (C) , and Alizarin Red stained calcium nodules (D) .
Figure Legend Snippet: Features and immunophenotypic mesenchymal profile of human-derived adipose mesenchymal stem cell (ADSCs). (A) Immunophenotypic mesenchymal profile of ADSCs shows that these cells express a high percentage of positivity for CD73, CD90, CD105, and a low-positivity fraction for CD14, CD19, CD34, CD45 and HLA-DR. After selective stimulation, ADSCs exhibited adipogenic, chondrogenic and osteogenic potential, which was demonstrated by Oil Red O staining of lipid droplets (B) , aggrecan deposition (C) , and Alizarin Red stained calcium nodules (D) .

Techniques Used: Derivative Assay, Staining

39) Product Images from "Calcium-Binding Proteins S100A8 and S100A9: Investigation of Their Immune Regulatory Effect in Myeloid Cells"

Article Title: Calcium-Binding Proteins S100A8 and S100A9: Investigation of Their Immune Regulatory Effect in Myeloid Cells

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19071833

S100A9-positive monocytes express HLA-DR. Peripheral blood mononuclear cells (PBMC) were stained for CD14, HLA-DR, and S100A9, and were gated on CD14+ cell populations. ( A ) Representative FACS plot showing that CD14+ monocytes express both HLA-DR and S100A9. Gates were set to the upper and lower 10% of HLA-DR expression for the CD14+ monocytes, to further study subsets; ( B , C ) S100A9 expression was not significant between HLA-DR-high and HLA-DR-low subsets, although, S100A9 was slightly higher in the latter subset. Significant differences were calculated by t -test; ( D ) Representative FACS plot showing expression of CD14 and HLA-DR in gated CD33+ myeloid cells; ( E ) Monocytic MDSC (CD14+ HLA-DR−) subsets showed slightly higher expression of S100A9 than CD14+ HLA-DR+ subsets, but this was not significant. The differences were tested by one-way ANOVA with Tukey’s multiple comparison tests. All data are expressed as means ± SD of four different experiments. * p
Figure Legend Snippet: S100A9-positive monocytes express HLA-DR. Peripheral blood mononuclear cells (PBMC) were stained for CD14, HLA-DR, and S100A9, and were gated on CD14+ cell populations. ( A ) Representative FACS plot showing that CD14+ monocytes express both HLA-DR and S100A9. Gates were set to the upper and lower 10% of HLA-DR expression for the CD14+ monocytes, to further study subsets; ( B , C ) S100A9 expression was not significant between HLA-DR-high and HLA-DR-low subsets, although, S100A9 was slightly higher in the latter subset. Significant differences were calculated by t -test; ( D ) Representative FACS plot showing expression of CD14 and HLA-DR in gated CD33+ myeloid cells; ( E ) Monocytic MDSC (CD14+ HLA-DR−) subsets showed slightly higher expression of S100A9 than CD14+ HLA-DR+ subsets, but this was not significant. The differences were tested by one-way ANOVA with Tukey’s multiple comparison tests. All data are expressed as means ± SD of four different experiments. * p

Techniques Used: Staining, FACS, Expressing

40) Product Images from "The roles of Wnt antagonists Dkk1 and sFRP4 during adipogenesis of human adipose tissue‐derived mesenchymal stem cells"

Article Title: The roles of Wnt antagonists Dkk1 and sFRP4 during adipogenesis of human adipose tissue‐derived mesenchymal stem cells

Journal: Cell Proliferation

doi: 10.1111/j.1365-2184.2008.00565.x

Characterization of isolated hAMSC. (a) Phase‐contrast microscopy of the spindle‐shape morphology of hAMSCs in the third passage. (b) Osteogenic differentiation of hAMSC: hAMSCs were cultured for 3 weeks in osteogenic induction medium, and calcium deposits were visualized via von Kossa staining. (c) Chondrogenic differentiation of hAMSC: hMASCs were cultured for 3 weeks in chondrogenic medium, and were stained with toluidine blue. (d) Adipogenic differentiation of hAMSC: hAMSCs were cultured for 3 weeks in adipogenic induction medium. Lipid droplets were visualized by oil red o staining. Bar represents 100 µm. (e) Neurogenic differentiation: immunostaining results for MAP2, GFAP and Tuj‐1. Bar represents 20 µm. (f) FACS analysis of hAMSC: the results showed that hAMSCs expressed CD90, CD105 and CD29 but did not express CD14, CD133, CD34, CD45 and HLA‐DR. The experiments were repeated three times.
Figure Legend Snippet: Characterization of isolated hAMSC. (a) Phase‐contrast microscopy of the spindle‐shape morphology of hAMSCs in the third passage. (b) Osteogenic differentiation of hAMSC: hAMSCs were cultured for 3 weeks in osteogenic induction medium, and calcium deposits were visualized via von Kossa staining. (c) Chondrogenic differentiation of hAMSC: hMASCs were cultured for 3 weeks in chondrogenic medium, and were stained with toluidine blue. (d) Adipogenic differentiation of hAMSC: hAMSCs were cultured for 3 weeks in adipogenic induction medium. Lipid droplets were visualized by oil red o staining. Bar represents 100 µm. (e) Neurogenic differentiation: immunostaining results for MAP2, GFAP and Tuj‐1. Bar represents 20 µm. (f) FACS analysis of hAMSC: the results showed that hAMSCs expressed CD90, CD105 and CD29 but did not express CD14, CD133, CD34, CD45 and HLA‐DR. The experiments were repeated three times.

Techniques Used: Isolation, Microscopy, Cell Culture, Staining, Immunostaining, FACS

Related Articles

other:

Article Title: Brain-Derived Neurotrophic Factor and Immune Cells in Osteoarthritis, Chronic Low Back Pain, and Chronic Widespread Pain Patients: Association with Anxiety and Depression
Article Snippet: The following antibodies were used for this study: CD3 PE-Cy7, CD4 FITC, CD8 PB, CD14 BV510, CD20 APC, CD45 APC-Cy, and CD56 PE.

Staining:

Article Title: Dysregulation of the Immune Environment in the Airways During HIV Infection
Article Snippet: .. Phenotyping by Multiparameter Flow CytometryThe staining panel consisted of CCR5 PE (2D7), CD38 APC (HIT2), CD3 PE-Cy7 (SK7), HLA-DR APC-Cy7 (L243; all from BD Biosciences, New Jersey, USA), CD4 PE-Cy5.5 (S3.5), CD8 Qdot-705 (3B5), CD19 Pacific Blue (SJ25-CI), CD14 Pacific Blue (T̈k4; all from Invitrogen, California, USA), CD45RO ECD (UCHL1), CD27 PE-Cy5 (1A4CD27; both from Beckman Coulter, California, USA). ..

Labeling:

Article Title: Expression of NK Cell Receptor Ligands on Leukemic Cells Is Associated with the Outcome of Childhood Acute Leukemia
Article Snippet: .. Briefly, 100µL of BM samples diluted with PBS-1%BSA to contain 0.5 million total white cells were labeled in 5 tubes, all of them containing CD45-APCCy7 (BD, Ref. 348815) and, depending on the type of leukemia, CD7-PECy7 (BD, Ref. 564019) for T-ALL, CD19-PECy7 (BD, Ref. 341113) for B-ALL or CD34 PE-Cy7 (BD, Ref. 348811) or CD33-PECy7 (BD, Ref. 333946) for AML, so as to detect total lymphocytes, granulocytes and leukemic blast cells following the gating strategy described in . ..

Activation Assay:

Article Title: Experimental postoperative ileus: is Th2 immune response involved?
Article Snippet: .. Leukocyte Activation Cocktail with BD GolgiPlug™, FACS antibodies include anti-mouse I-A/I-E-BV510, IgG1-BB700, IgM-BV605, IgE-BV786, IgD-BV711, CD1d-BV421, CD5-PE, CD45-BUV395, CD19-APC, CD45R/B220-BUV496, CD45-APC-Cy7, CD3e-FITC, CD4-V450, CD8-BV510, CD25-BV605, IL-4-PE-Cy7, IFN-γ-PE, FoxP3-AF647, CD103-BUV395, F4/80-BV711, CD80-BV650, CD11b-BV510, Ly6-G-PerCP Cy5.5, PE-Ly6-C, CD45-APC-Cy7, CD11c--AF700 were purchased from BD (Heidelberg, Germany); Another FACS antibody Anti-mouse-IL-17A-BV650 was purchased from eBioscience (Frankfurt am Main, Germany). ..

FACS:

Article Title: Experimental postoperative ileus: is Th2 immune response involved?
Article Snippet: .. Leukocyte Activation Cocktail with BD GolgiPlug™, FACS antibodies include anti-mouse I-A/I-E-BV510, IgG1-BB700, IgM-BV605, IgE-BV786, IgD-BV711, CD1d-BV421, CD5-PE, CD45-BUV395, CD19-APC, CD45R/B220-BUV496, CD45-APC-Cy7, CD3e-FITC, CD4-V450, CD8-BV510, CD25-BV605, IL-4-PE-Cy7, IFN-γ-PE, FoxP3-AF647, CD103-BUV395, F4/80-BV711, CD80-BV650, CD11b-BV510, Ly6-G-PerCP Cy5.5, PE-Ly6-C, CD45-APC-Cy7, CD11c--AF700 were purchased from BD (Heidelberg, Germany); Another FACS antibody Anti-mouse-IL-17A-BV650 was purchased from eBioscience (Frankfurt am Main, Germany). ..

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Becton Dickinson hla dr
    Elevated percentage of classic monocytes in patients with axial spondyloarthritis (axSpA). a The percentage of classic <t>CD14</t> ++ CD16 − (M1), intermediate CD14 ++ CD16 + (M2), and non-classic CD14 + CD16 + (M3) monocytes was determined by fluorescence-activated cell sorting in whole blood of controls, patients with axSpA, and patients with rheumatoid arthritis (RA). The percentage of classic, intermediate, and non-classic monocytes was determined among monocytes which were gated according to forward (FSC) and side (SSC) scatter (R1) and <t>HLA-DR</t> expression (R2). b The percentage of classic, intermediate, and non-classic monocytes (individual measurements and median) is shown from measurements in 12 controls (C), 20 patients with axSpA (SpA), and 12 patients with RA (RA). * P
    Hla Dr, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hla dr/product/Becton Dickinson
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hla dr - by Bioz Stars, 2021-09
    97/100 stars
      Buy from Supplier

    86
    Becton Dickinson human leucocyte antigen d related hla dr
    Confocal analysis of CD1a and human leucocyte antigen D-related <t>(HLA-DR)</t> expression in monocyte-derived immature dendritic cells (iDC). (a) Images of monocyte-derived DC obtained by confocal microscopy with CD1a in green, HLA-DR in red and nucleus in
    Human Leucocyte Antigen D Related Hla Dr, 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
    https://www.bioz.com/result/human leucocyte antigen d related hla dr/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human leucocyte antigen d related hla dr - by Bioz Stars, 2021-09
    86/100 stars
      Buy from Supplier

    86
    Becton Dickinson hla dr pe
    Hypoxia potentiates LPS-induced TNF-α secretion ( A – C ) Cell viability of moDCs during atmospheric (20% O 2 ) and hypoxic (1% O 2 ) oxygen levels in presence or absence of LPS, determined by Zombie Violet as analyzed by flow cytometry. Representative dot plot (A), histograms (B), and quantitation for three donors ((C); average ± S.E.M.) are shown. ( D – I ) LPS-induced maturation of moDCs cultured at atmospheric or hypoxic oxygen levels, determined by expression levels of maturation markers <t>CD83</t> (D,G), CD86 (E,H), and <t>HLA-DR</t> (F,I). Representative histograms showed quantitation for five donors. ( J , K ) Secretion by ELISA of the pro-inflammatory cytokines TNF-α (J) and IL-6 (K) from moDCs cultured at atmospheric or hypoxic oxygen levels. Quantitation from ten donors (average ± S.E.M.). * P
    Hla Dr Pe, 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
    https://www.bioz.com/result/hla dr pe/product/Becton Dickinson
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hla dr pe - by Bioz Stars, 2021-09
    86/100 stars
      Buy from Supplier

    Image Search Results


    Elevated percentage of classic monocytes in patients with axial spondyloarthritis (axSpA). a The percentage of classic CD14 ++ CD16 − (M1), intermediate CD14 ++ CD16 + (M2), and non-classic CD14 + CD16 + (M3) monocytes was determined by fluorescence-activated cell sorting in whole blood of controls, patients with axSpA, and patients with rheumatoid arthritis (RA). The percentage of classic, intermediate, and non-classic monocytes was determined among monocytes which were gated according to forward (FSC) and side (SSC) scatter (R1) and HLA-DR expression (R2). b The percentage of classic, intermediate, and non-classic monocytes (individual measurements and median) is shown from measurements in 12 controls (C), 20 patients with axSpA (SpA), and 12 patients with RA (RA). * P

    Journal: Arthritis Research & Therapy

    Article Title: In vivo pre-activation of monocytes in patients with axial spondyloarthritis

    doi: 10.1186/s13075-015-0694-2

    Figure Lengend Snippet: Elevated percentage of classic monocytes in patients with axial spondyloarthritis (axSpA). a The percentage of classic CD14 ++ CD16 − (M1), intermediate CD14 ++ CD16 + (M2), and non-classic CD14 + CD16 + (M3) monocytes was determined by fluorescence-activated cell sorting in whole blood of controls, patients with axSpA, and patients with rheumatoid arthritis (RA). The percentage of classic, intermediate, and non-classic monocytes was determined among monocytes which were gated according to forward (FSC) and side (SSC) scatter (R1) and HLA-DR expression (R2). b The percentage of classic, intermediate, and non-classic monocytes (individual measurements and median) is shown from measurements in 12 controls (C), 20 patients with axSpA (SpA), and 12 patients with RA (RA). * P

    Article Snippet: The remaining cells were stained with the respective antibodies against CD14 (clone M5E2 PerCPcy5.5; BD), CD16 (clone eBioCB16 APC; ebioscience, Frankfurt, Germany), HLA-DR (clone L243 fluorescein isothiocyanate (FITC); BD Heidelberg, Germany), CD64 (clone 10.1 PE; Dako, Glostrup, DK), CD80 (clone 2D10 PE; biolegend, San Diego, CA, USA), CD115 (clone 12-3A3-1B10 PE; ebioscience), and CD163 (clone GHI/61PE; BD).

    Techniques: Fluorescence, FACS, Expressing

    UC-MSC phenotype analysis. MSCs derived from umbilical cord were detached with trypsin-EDTA and stained with fluorescence antibodies against surface molecules indicated and analyzed by flow cytometry present in histogram plots. Appropriate isotype controls were performed (for each antibody isotype) to assess cell auto-fluorescence and background staining. MSCs were positive for CD73 (99%), CD90 (99%), CD105 (95%), CD166 (98%), negative for hematopoietic marker CD34 (1.7%), leukocyte common antigen CD45 (2.2%), HLA-DR (1.3%) and monocyte marker CD14 (1.5%).

    Journal: Journal of Translational Medicine

    Article Title: Laminin 411 acts as a potent inducer of umbilical cord mesenchymal stem cell differentiation into insulin-producing cells

    doi: 10.1186/1479-5876-12-135

    Figure Lengend Snippet: UC-MSC phenotype analysis. MSCs derived from umbilical cord were detached with trypsin-EDTA and stained with fluorescence antibodies against surface molecules indicated and analyzed by flow cytometry present in histogram plots. Appropriate isotype controls were performed (for each antibody isotype) to assess cell auto-fluorescence and background staining. MSCs were positive for CD73 (99%), CD90 (99%), CD105 (95%), CD166 (98%), negative for hematopoietic marker CD34 (1.7%), leukocyte common antigen CD45 (2.2%), HLA-DR (1.3%) and monocyte marker CD14 (1.5%).

    Article Snippet: UC-MSCs were resuspended in PBS, stained with fluorescence-labeled monoclonal antibodies CD34, HLA-DR, CD105, CD73, CD90, and CD45 (BD, Franklin Lakes, NY, USA), and then incubated for 30 min at 4°C in the dark.

    Techniques: Derivative Assay, Staining, Fluorescence, Flow Cytometry, Cytometry, Marker

    Confocal analysis of CD1a and human leucocyte antigen D-related (HLA-DR) expression in monocyte-derived immature dendritic cells (iDC). (a) Images of monocyte-derived DC obtained by confocal microscopy with CD1a in green, HLA-DR in red and nucleus in

    Journal: Clinical and Experimental Immunology

    Article Title: Alteration of CD1 expression in multiple sclerosis

    doi: 10.1111/j.1365-2249.2012.04586.x

    Figure Lengend Snippet: Confocal analysis of CD1a and human leucocyte antigen D-related (HLA-DR) expression in monocyte-derived immature dendritic cells (iDC). (a) Images of monocyte-derived DC obtained by confocal microscopy with CD1a in green, HLA-DR in red and nucleus in

    Article Snippet: Cells were stained for 30 min at 4°C with the following fluorochrome-conjugated mouse monoclonal antibodies (mAbs): CD1a (clone HI149), CD1b (clone M-T101), CD1d (clone CD1d42), human leucocyte antigen D-related (HLA-DR) (clone L243), CD14 (clone MjP9) and CD209 (clone DCN46) obtained from BD-Pharmingen (San Diego, CA, USA) and CD1c (BDCA-1) from Miltenyi Biotec (Bergisch Gladbach, Germany).

    Techniques: Expressing, Derivative Assay, Confocal Microscopy

    Hypoxia potentiates LPS-induced TNF-α secretion ( A – C ) Cell viability of moDCs during atmospheric (20% O 2 ) and hypoxic (1% O 2 ) oxygen levels in presence or absence of LPS, determined by Zombie Violet as analyzed by flow cytometry. Representative dot plot (A), histograms (B), and quantitation for three donors ((C); average ± S.E.M.) are shown. ( D – I ) LPS-induced maturation of moDCs cultured at atmospheric or hypoxic oxygen levels, determined by expression levels of maturation markers CD83 (D,G), CD86 (E,H), and HLA-DR (F,I). Representative histograms showed quantitation for five donors. ( J , K ) Secretion by ELISA of the pro-inflammatory cytokines TNF-α (J) and IL-6 (K) from moDCs cultured at atmospheric or hypoxic oxygen levels. Quantitation from ten donors (average ± S.E.M.). * P

    Journal: Bioscience Reports

    Article Title: Hypoxia potentiates monocyte-derived dendritic cells for release of tumor necrosis factor α via MAP3K8

    doi: 10.1042/BSR20182019

    Figure Lengend Snippet: Hypoxia potentiates LPS-induced TNF-α secretion ( A – C ) Cell viability of moDCs during atmospheric (20% O 2 ) and hypoxic (1% O 2 ) oxygen levels in presence or absence of LPS, determined by Zombie Violet as analyzed by flow cytometry. Representative dot plot (A), histograms (B), and quantitation for three donors ((C); average ± S.E.M.) are shown. ( D – I ) LPS-induced maturation of moDCs cultured at atmospheric or hypoxic oxygen levels, determined by expression levels of maturation markers CD83 (D,G), CD86 (E,H), and HLA-DR (F,I). Representative histograms showed quantitation for five donors. ( J , K ) Secretion by ELISA of the pro-inflammatory cytokines TNF-α (J) and IL-6 (K) from moDCs cultured at atmospheric or hypoxic oxygen levels. Quantitation from ten donors (average ± S.E.M.). * P

    Article Snippet: The following directly labeled antibodies were used: CD11c-FITC (BD Biosciences, Franklin Lakes, U.S.A.), CD14-PE-Cy7 (Miltenyi Biotec, Bergisch Gladbach, Germany), CD16-APC (Miltenyi Biotec), CD11b-PE-Cy7 (Beckman Coulter, Brea, U.S.A.), CD68-APC (BioLegend), HLA-DR-PE (BD Biosciences), CD83-FITC (BD Biosciences), CD86-APC (BD Biosciences), TLR2-FITC (BioLegend), TLR4-APC (BioLegend).

    Techniques: Flow Cytometry, Cytometry, Quantitation Assay, Cell Culture, Expressing, Enzyme-linked Immunosorbent Assay