Journal: Mediators of Inflammation

Article Title: Regulatory Effect of Iguratimod on the Balance of Th Subsets and Inhibition of Inflammatory Cytokines in Patients with Rheumatoid Arthritis

doi: 10.1155/2015/356040

Figure Lengend Snippet: Frequencies of CD4 + T cell subset in RA patients before and after therapy. The proportion of IFN- γ + Th1, IL-17 + Th17, circulating CD4 + CCR7 low PD-1 high Tfh, and CD4 + CD25 + Foxp3 + Treg in CD4 + T cells was analyzed by flow cytometry on 0 w, 24 w, and 52 w. (a) Representative staining data showing the proportion of Th1. (b) Representative staining data showing the proportion of Th17. (c) Representative staining data showing the proportion of Tfh. (d) Representative staining data showing the proportion of Treg. (e) Collated data showing the proportion of Th1. (f) Collated data showing the proportion of Th17. (g) Collated data showing the proportion of Tfh. (h) Collated data showing the proportion of Treg. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 are statistically significant compared to 24 w. # p < 0.05, ## p < 0.01, and ### p < 0.001 are statistically significant compared to 0 w. Group A: iguratimod was administered to patients for 52 weeks. Group B: placebo was administered to patients for the first 24 weeks and iguratimod was administered to patients for the subsequent 28 weeks.

Article Snippet: For Tfh analysis, cells were incubated with Percp cy5.5-conjugated anti-human CD4, FITC-conjugated anti-CCR7 (BD Pharmingen, San Diego, CA, USA) and APC-conjugated anti-PD-1 (BD Pharmingen, San Diego, CA, USA).

Techniques: Flow Cytometry, Staining

Journal: Cancer Management and Research

Article Title: Programmed Cell Death-1 and Its Ligands as Targets for Therapy of Multiple Myeloma Patients

doi: 10.2147/CMAR.S351383

Figure Lengend Snippet: Median Expression of PDCD1, PDCD1LG1, PDCD1LG2 and Their Splicing Variants

Article Snippet: The combination of fluorescent-labeled monoclonal antibodies (Mabs) was used as follows: anti-PD-1-allophycocyanin (APC) (BD Biosciences, San Jose, CA, USA), anti-PD-L1-APC (BD Biosciences, San Jose, CA, USA), anti-PD-L2-APC (BD Biosciences, San Jose, CA, USA), anti-CD45-brilliant violet 421 (BV421) (BD Biosciences, San Jose, CA, USA), anti-CD56-BV510 (BD Biosciences, San Jose, CA, USA), anti-CD19-peridinin-chlorophyll proteins (PerCP)-Cy5.5 (BD Biosciences, San Jose, CA, USA), anti-CD138-fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA, USA), and anti-CD38-phycoerythrin (PE) (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Variant Assay

Journal: Cancer Management and Research

Article Title: Programmed Cell Death-1 and Its Ligands as Targets for Therapy of Multiple Myeloma Patients

doi: 10.2147/CMAR.S351383

Figure Lengend Snippet: The surface expression of PD-1 on plasma cells and B cells in BM (N=98) of MM patients and B cells derived from PB of HVs (N=10). The central line shows the median.

Article Snippet: The combination of fluorescent-labeled monoclonal antibodies (Mabs) was used as follows: anti-PD-1-allophycocyanin (APC) (BD Biosciences, San Jose, CA, USA), anti-PD-L1-APC (BD Biosciences, San Jose, CA, USA), anti-PD-L2-APC (BD Biosciences, San Jose, CA, USA), anti-CD45-brilliant violet 421 (BV421) (BD Biosciences, San Jose, CA, USA), anti-CD56-BV510 (BD Biosciences, San Jose, CA, USA), anti-CD19-peridinin-chlorophyll proteins (PerCP)-Cy5.5 (BD Biosciences, San Jose, CA, USA), anti-CD138-fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA, USA), and anti-CD38-phycoerythrin (PE) (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Derivative Assay

Journal: Cancer Management and Research

Article Title: Programmed Cell Death-1 and Its Ligands as Targets for Therapy of Multiple Myeloma Patients

doi: 10.2147/CMAR.S351383

Figure Lengend Snippet: The comparison of PD-1 expression on CD4+ and CD8+ T cells in BM (N=45) ( B ) and PB (N=21) ( A ) of MM patients. The central line shows the median.

Article Snippet: The combination of fluorescent-labeled monoclonal antibodies (Mabs) was used as follows: anti-PD-1-allophycocyanin (APC) (BD Biosciences, San Jose, CA, USA), anti-PD-L1-APC (BD Biosciences, San Jose, CA, USA), anti-PD-L2-APC (BD Biosciences, San Jose, CA, USA), anti-CD45-brilliant violet 421 (BV421) (BD Biosciences, San Jose, CA, USA), anti-CD56-BV510 (BD Biosciences, San Jose, CA, USA), anti-CD19-peridinin-chlorophyll proteins (PerCP)-Cy5.5 (BD Biosciences, San Jose, CA, USA), anti-CD138-fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA, USA), and anti-CD38-phycoerythrin (PE) (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Programmed Cell Death-1 and Its Ligands as Targets for Therapy of Multiple Myeloma Patients

doi: 10.2147/CMAR.S351383

Figure Lengend Snippet: The surface expression of PD-1 on CD8+ ( A ) and CD4+ ( B ) T cells in PB (N=21) and BM (n=45) of MM patients and HVs (N=30). The central line shows the median.

Article Snippet: The combination of fluorescent-labeled monoclonal antibodies (Mabs) was used as follows: anti-PD-1-allophycocyanin (APC) (BD Biosciences, San Jose, CA, USA), anti-PD-L1-APC (BD Biosciences, San Jose, CA, USA), anti-PD-L2-APC (BD Biosciences, San Jose, CA, USA), anti-CD45-brilliant violet 421 (BV421) (BD Biosciences, San Jose, CA, USA), anti-CD56-BV510 (BD Biosciences, San Jose, CA, USA), anti-CD19-peridinin-chlorophyll proteins (PerCP)-Cy5.5 (BD Biosciences, San Jose, CA, USA), anti-CD138-fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA, USA), and anti-CD38-phycoerythrin (PE) (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: Programmed Cell Death-1 and Its Ligands as Targets for Therapy of Multiple Myeloma Patients

doi: 10.2147/CMAR.S351383

Figure Lengend Snippet: The comparison of PD-1-expression T cells determined by mean fluorescence intensity (MFI) in peripheral blood of RRMM patients before and after 5 cycles of daratumumab treatment. The central line shows the median.

Article Snippet: The combination of fluorescent-labeled monoclonal antibodies (Mabs) was used as follows: anti-PD-1-allophycocyanin (APC) (BD Biosciences, San Jose, CA, USA), anti-PD-L1-APC (BD Biosciences, San Jose, CA, USA), anti-PD-L2-APC (BD Biosciences, San Jose, CA, USA), anti-CD45-brilliant violet 421 (BV421) (BD Biosciences, San Jose, CA, USA), anti-CD56-BV510 (BD Biosciences, San Jose, CA, USA), anti-CD19-peridinin-chlorophyll proteins (PerCP)-Cy5.5 (BD Biosciences, San Jose, CA, USA), anti-CD138-fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA, USA), and anti-CD38-phycoerythrin (PE) (BD Biosciences, San Jose, CA, USA).

Techniques: Expressing, Fluorescence

Journal: Clinical and Developmental Immunology

Article Title: Cytotoxic Activity of Dendritic Cells as a Possible Mechanism of Negative Regulation of T Lymphocytes in Pulmonary Tuberculosis

doi: 10.1155/2012/628635

Figure Lengend Snippet: The number of freshly isolated PD-1 + CD4 + and PD-1 + CD8 + cells in PPD-anergic TB patients was higher than that of healthy donors. The numbers of PD-1 + CD4 + (M ± S.E.) cells (a) and of PD-1 + CD8 + cells (b) were detected in the peripheral blood of healthy donors: PPD-reactive and PPD-anergic TB patients.

Article Snippet: To evaluate PD-1 expression on CD4 + and CD8 + T-lymphocytes, MNCs (either freshly-isolated or stimulated in allo-MLC) were incubated with the following antibodies: FITC-labeled anti-CD3, PE-labeled anti-CD4 and anti-CD8 (Sorbent, Moscow), and APC-labeled anti-PD-1 (Becton Dickinson).

Techniques: Isolation

Journal: Clinical and Developmental Immunology

Article Title: Cytotoxic Activity of Dendritic Cells as a Possible Mechanism of Negative Regulation of T Lymphocytes in Pulmonary Tuberculosis

doi: 10.1155/2012/628635

Figure Lengend Snippet: Addition of neutralizing anti-PD-1 antibodies in 3-day MLC stimulated with healthy donor and TB patient DCs was accompanied by decrease in CD3 + CD4 + and CD3 + CD8 + T-cell apoptosis. The number of apoptotic CD3 + CD4 + (M ± S.E.) (a) and CD3 + CD8 + (b) T-cells was determined in allo-MLC stimulated with healthy donor and TB patient DCs in the absence and presence of anti-PD-1 antibodies (5 μ g/mL).

Article Snippet: To evaluate PD-1 expression on CD4 + and CD8 + T-lymphocytes, MNCs (either freshly-isolated or stimulated in allo-MLC) were incubated with the following antibodies: FITC-labeled anti-CD3, PE-labeled anti-CD4 and anti-CD8 (Sorbent, Moscow), and APC-labeled anti-PD-1 (Becton Dickinson).

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: CD4 + CXCR5 + PD-1 + T Follicular Helper Cells Play a Pivotal Role in the Development of Rheumatoid Arthritis

doi: 10.12659/MSM.914868

Figure Lengend Snippet: Frequency of circulating follicular helper T (Tfh) cells was increased and associated positively with the levels of autoantibodies and inflammation markers in new-onset rheumatoid arthritis (RA) patients. Peripheral blood mononuclear cells (PBMCs) from RA patients and healthy controls were stained with labelled antibodies as previously described, and analyzed by flow cytometry. ( A ) Percentage of CD4+ T lymphocytes in RA patients and healthy controls. ( B ) Percentage of CD4+ CXCR5+ PD-1+ T cells in RA patients and healthy controls (* P <0.01). ( C ) The percentage of Tfh cells was correlated positively with the level of serum RF (rheumatoid factor) in new-onset RA (r=0.962, P <0.0001). ( D ) The percentage of Tfh cells was correlated positively with the level of serum ACPA (anti-cyclic citrullinated peptide antibodies) in new-onset RA (r=0.966, P <0.0001). ( E ) The percentage of Tfh cells was correlated positively with the value of ESR (erythrocyte sedimentation rate) in new-onset RA (r=0.968, P <0.0001). ( F ) The percentage of Tfh cells was correlated positively with the level of serum CRP (C-reactive protein) in new-onset RA (r=0.953, P <0.0001).

Article Snippet: PBMCs at 2×10 6 /tube were stained with Percpcy5.5 anti-human CD4 (Cat. No. 560650), AF488 anti-human CXCR5 (Cat. No. 558112), and APC anti-human PD-1 (Cat. No. 558694) (BD Bioscience, San Diego, CA, USA).

Techniques: Staining, Flow Cytometry, Sedimentation

Journal: Frontiers in Physiology

Article Title: Regulatory Effect of Sishen Pill on Tfh Cells in Mice With Experimental Colitis

doi: 10.3389/fphys.2020.00589

Figure Lengend Snippet: Flow cytometry to detect CD4 + CXCR5 + BCL-6 + , CD4 + CXCR5 + Blimp-1 + , CD4 + CXCR5 + PD-1 + , and CD4 + CXCR5 + PD-L1 + cell levels. (A) Total lymphocytes in peripheral blood. (B) Gating strategy for CD4 + cells. (C) C1–C5: CD4 + CXCR5 + BCL-6 + cell levels. (D) D1–D5: CD4 + CXCR5 + Blimp-1 + cell levels. (E) E1–E5: CD4 + CXCR5 + PD-1 + cell levels. (F) F1–F5: CD4 + CXCR5 + PD-L1 + cell level. Data were presented as means ± SEM ( n = 8). # p < 0.05 and ## p < 0.01 versus the normal group; * p < 0.05 and ** p < 0.01 versus the DSS group.

Article Snippet: The following monoclonal antibodies were used: APC-H7 anti-mouse CD4 (No. 560181) (1:200), FITC Anti-Mouse CXCR5 (No. 560577) (1:100), PE Anti-Mouse ICOS (No. 565669) (1:100), PerCP-Cy5.5 Rat Anti-Mouse FOXP3 (No. 563902) (1:100), APC Anti-Mouse IL-10 (No. 554468) (1:200), APC-Cy7 Anti-Mouse IL-17 (No. 560821) (1:200), APC Anti-Mouse PD-1 (No. 562671) (1:200), PE Anti-Mouse PD-L1 (No. 558091) (1:200), and PerCP-Cy5.5 Anti-Mouse BCL-6 (No. 563582) (1:200) (BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Flow Cytometry

Journal: Leukemia

Article Title: Expression of PD-L1, PD-L2, PD-1 and CTLA4 in myelodysplastic syndromes is enhanced by treatment with hypomethylating agents

doi: 10.1038/leu.2013.355

Figure Lengend Snippet: (A) Increased mRNA expression of PD-L1, PD-L2, PD-1 and CTLA4 genes in CD34+ cells from MDS, CMML and AML patients by real-time PCR analysis, normal CD34+ cells were used as normal control. N = number of patients. MDS, myelodysplastic syndromes; CMML, chronic myelomonocytic leukemia; AML, acute myeloid leukemia. (B) Increased mRNA expression of PD-L1, PD-L2, PD-1 and CTLA4 in PBMNCs from MDS, CMML and AML patients, normal PBMNCs were used as normal controls. PBMNC, peripheral blood mononuclear cells. (C) Comparisons of mRNA expression levels of PD-L1, PD-L2, PD-1 and CTLA4 between CD34+ cells and PBMNC from MDS, CMML and AML patients.

Article Snippet: APC mouse anti-human PD-1 and PE mouse anti-human PD-L1 antibodies were obtained from BD Biosciences (San Jose, CA).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Leukemia

Article Title: Expression of PD-L1, PD-L2, PD-1 and CTLA4 in myelodysplastic syndromes is enhanced by treatment with hypomethylating agents

doi: 10.1038/leu.2013.355

Figure Lengend Snippet: (A) Illustrations of PD-L1 membranous expression in bone marrow biopsy samples from MDS, CMML and AML patients (X1000): a, Representative case show strong PD-L1 membranous expression in blasts; b, Representative case show negative PD-L1 membranous expression in blasts. (B) Illustrations of PD-1 membranous expression in bone marrow biopsy samples from MDS, CMML and AML patients (X1000): a, Representative case show PD-1 membranous expression in blasts; b, Representative case show negative PD-1 membranous expression in blasts, but positive PD-1 expression in stroma/non-blast compartment. (C) Illustrations of PD-L1 membranous expression in CD34+ cell cytospin from MDS, CMML and AML patients (X1000): a, Representative case show strong PD-L1 membranous expression; b, Representative case show negative PD-L1 membranous expression. (D) Illustrations of PD-1 membranous expression in CD34+ cell cytospin from MDS, CMML and AML patients (X1000): a, Representative case show partial PD-1 membranous expression; b, Representative case show negative PD-1 membranous expression.

Article Snippet: APC mouse anti-human PD-1 and PE mouse anti-human PD-L1 antibodies were obtained from BD Biosciences (San Jose, CA).

Techniques: Expressing

Journal: Leukemia

Article Title: Expression of PD-L1, PD-L2, PD-1 and CTLA4 in myelodysplastic syndromes is enhanced by treatment with hypomethylating agents

doi: 10.1038/leu.2013.355

Figure Lengend Snippet: (A) Summary of dynamics of PD-L1, PD-L2, PD-1 and CTLA4 mRNA expression in 61 patients treated with different forms of epigenetic therapy, expression on day 0 was baseline expression before treatment, most patients received 5 days of hypomethylating agent treatment. (B) Induction of PD-L1, PD-L2, PD-1 and CTLA4 expression in patients treated in a phase 2 trial of vorinostat in combination with azacitidine. C=course, D=days on therapy. Resistance: patients had no response to treatment; Responing: patients acquired complete remission after treatment. (C) Overall survival by comparison of PD-L2 upregulation after treatment in group of patients from a phase 2 trial of vorinostat in combination with azacitidine. Group 0: group of patients without PD-L2 expression induction (≥2 fold) after theatment; Group 1: group of patients acquired PD-L2 expression induction (≥2 fold) after theatment.

Article Snippet: APC mouse anti-human PD-1 and PE mouse anti-human PD-L1 antibodies were obtained from BD Biosciences (San Jose, CA).

Techniques: Expressing

Journal: Leukemia

Article Title: Expression of PD-L1, PD-L2, PD-1 and CTLA4 in myelodysplastic syndromes is enhanced by treatment with hypomethylating agents

doi: 10.1038/leu.2013.355

Figure Lengend Snippet: (A) Induction of PD-L1, PD-1 and CTLA4 mRNA expression in KG-1 treated with different concentrations of decitabine and cytarabine. (B) Induction of PD-L1 and PD-L2 mRNA expression in THP1 treated with different concentrations of decitabine and cytarabine. (C) Flow cytometry analysis of PD-1 and PD-L1 protein expression levels in KG-1 treated with different concentrations of decitabine.

Article Snippet: APC mouse anti-human PD-1 and PE mouse anti-human PD-L1 antibodies were obtained from BD Biosciences (San Jose, CA).

Techniques: Expressing, Flow Cytometry

Journal: Leukemia

Article Title: Expression of PD-L1, PD-L2, PD-1 and CTLA4 in myelodysplastic syndromes is enhanced by treatment with hypomethylating agents

doi: 10.1038/leu.2013.355

Figure Lengend Snippet: (A) PD-1 methylation in leukemia cell lines, AML patients and normal controls. (B) Dynamics of PD-1 methylation in KG-1 leukemia cell line treated with decitabine by pyrosequencing analysis, N=Number. (C) Bisulfite sequencing analysis of dynamics of PD-1 methylation with decitabine treatment of KG-1. (D) Dynamics of PD-1 methylation in AML and MDS patients from group of patients treated with vorinostat in combination with azacitidine, C=Course; D=days on therapy.

Article Snippet: APC mouse anti-human PD-1 and PE mouse anti-human PD-L1 antibodies were obtained from BD Biosciences (San Jose, CA).

Techniques: Methylation, Methylation Sequencing