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

Article Title: Costimulatory pathways in multiple sclerosis: distinctive expression of PD-1 and PD-L1 in patients with different patterns of disease.

doi: 10.4049/jimmunol.0901038

Figure Lengend Snippet: FIGURE 1. MBP-stimulated CD4 and CD8 expressing PD-1. Representative results of MBP-stimulated PBMC of patients with either AMS or SMS are shown. Top panels, PD-1-expressing MBP-stimulated CD4 T lymphocytes. Bottom panels, PD-1-expressing MBP-stimulated CD8 T lymphocytes. In the upper right corners, the percentage of CD4/PD-1 and CD8/PD-1 T cells relative to the total number of lymphocytes are indicated. Summary results obtained in AMS and SMS patients are presented in A and B. The boxes stretch from the 25th to the 75th percentile; the lines across the boxes indicate the median values; the lines stretching from the boxes indicate extreme values. Statistical significance is shown.

Article Snippet: The following mAbs were used in this study: anti-human CD4 (clone 13B8.2; mouse IgG2a isotype), anti-human CD14 (clone 116; mouse IgG1 isotype), anti-human CD19 (clone J4.119; mouse IgG1 isotype) coupled to R-PE-Cyanine 5 Tandem (PE-Cy5; Caltag Laboratories); anti-human CD8 (clone SFCI21thy2D3; mouse IgG1 isotype) PE-Cyanin-7 (BeckmanCoulter); anti-human CD86 (clone BU63; mouse IgG1 isotype), antihuman CD80 (clone 3H5; mouse IgG1 isotype; Serotec) coupled to FITC; anti-human PD-L1 (gift from Dr. L. Chen) and anti-mouse IgG (H L chain) coupled to FITC (eBioscience); anti-human-B7H3 (gift from Dr. L. Chen) coupled FITC anti-human PD-1 (clone MIH4) coupled to PE (mouse-IgG1 isotype; eBioscience); and anti-human annexin V coupled to FITC (mouse-IgG1 isotype; Beckman Coulter).

Techniques: Expressing

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

Article Title: Costimulatory pathways in multiple sclerosis: distinctive expression of PD-1 and PD-L1 in patients with different patterns of disease.

doi: 10.4049/jimmunol.0901038

Figure Lengend Snippet: FIGURE 4. Annexin V- and PD-1-expressing MBP-stimulated CD4 and CD8 T lymphocytes. Representative results of MBP-stimulated PBMC of patients with either AMS or SMS are shown. Top panels, MBP-stimulated CD4 T lymphocytes expressing annexin V and PD-1. Bottom panels, MBP-stimulated CD8 T lymphocytes expressing annexin V and PD-1. Summary results obtained in AMS and SMS patients are presented in A and B. The boxes stretch from the 25th to the 75th percentile; the lines across the boxes indicate the median values; the lines stretching from the boxes indicate extreme values. Statistical significance is shown.

Article Snippet: The following mAbs were used in this study: anti-human CD4 (clone 13B8.2; mouse IgG2a isotype), anti-human CD14 (clone 116; mouse IgG1 isotype), anti-human CD19 (clone J4.119; mouse IgG1 isotype) coupled to R-PE-Cyanine 5 Tandem (PE-Cy5; Caltag Laboratories); anti-human CD8 (clone SFCI21thy2D3; mouse IgG1 isotype) PE-Cyanin-7 (BeckmanCoulter); anti-human CD86 (clone BU63; mouse IgG1 isotype), antihuman CD80 (clone 3H5; mouse IgG1 isotype; Serotec) coupled to FITC; anti-human PD-L1 (gift from Dr. L. Chen) and anti-mouse IgG (H L chain) coupled to FITC (eBioscience); anti-human-B7H3 (gift from Dr. L. Chen) coupled FITC anti-human PD-1 (clone MIH4) coupled to PE (mouse-IgG1 isotype; eBioscience); and anti-human annexin V coupled to FITC (mouse-IgG1 isotype; Beckman Coulter).

Techniques: Expressing

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

Article Title: Costimulatory pathways in multiple sclerosis: distinctive expression of PD-1 and PD-L1 in patients with different patterns of disease.

doi: 10.4049/jimmunol.0901038

Figure Lengend Snippet: FIGURE 5. pAkt-expressing MBP-stimulated CD4 and CD8 T lymphocytes. Representative results of MBP-stimulated PBMC of patients with either AMS or stable SMS are shown. Top panels, MBP-stimulated CD4 T lymphocytes expressing pAkt. Bottom panels, MBP-stimulated CD8 T lymphocytes expressing pAkt. Summary results obtained in AMS and SMS patients are presented in A and B. The boxes stretch from the 25th to the 75th percentile; the lines across the boxes indicate the median values; the lines stretching from the boxes indicate extreme values. Statistical significance is shown.

Article Snippet: The following mAbs were used in this study: anti-human CD4 (clone 13B8.2; mouse IgG2a isotype), anti-human CD14 (clone 116; mouse IgG1 isotype), anti-human CD19 (clone J4.119; mouse IgG1 isotype) coupled to R-PE-Cyanine 5 Tandem (PE-Cy5; Caltag Laboratories); anti-human CD8 (clone SFCI21thy2D3; mouse IgG1 isotype) PE-Cyanin-7 (BeckmanCoulter); anti-human CD86 (clone BU63; mouse IgG1 isotype), antihuman CD80 (clone 3H5; mouse IgG1 isotype; Serotec) coupled to FITC; anti-human PD-L1 (gift from Dr. L. Chen) and anti-mouse IgG (H L chain) coupled to FITC (eBioscience); anti-human-B7H3 (gift from Dr. L. Chen) coupled FITC anti-human PD-1 (clone MIH4) coupled to PE (mouse-IgG1 isotype; eBioscience); and anti-human annexin V coupled to FITC (mouse-IgG1 isotype; Beckman Coulter).

Techniques: Expressing

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

Article Title: Costimulatory pathways in multiple sclerosis: distinctive expression of PD-1 and PD-L1 in patients with different patterns of disease.

doi: 10.4049/jimmunol.0901038

Figure Lengend Snippet: FIGURE 6. Blockade of the PD-1-PD-L1 pathway. pAKT-expressing, MBP-stimulated CD4 and CD8 T lymphocytes.

Article Snippet: The following mAbs were used in this study: anti-human CD4 (clone 13B8.2; mouse IgG2a isotype), anti-human CD14 (clone 116; mouse IgG1 isotype), anti-human CD19 (clone J4.119; mouse IgG1 isotype) coupled to R-PE-Cyanine 5 Tandem (PE-Cy5; Caltag Laboratories); anti-human CD8 (clone SFCI21thy2D3; mouse IgG1 isotype) PE-Cyanin-7 (BeckmanCoulter); anti-human CD86 (clone BU63; mouse IgG1 isotype), antihuman CD80 (clone 3H5; mouse IgG1 isotype; Serotec) coupled to FITC; anti-human PD-L1 (gift from Dr. L. Chen) and anti-mouse IgG (H L chain) coupled to FITC (eBioscience); anti-human-B7H3 (gift from Dr. L. Chen) coupled FITC anti-human PD-1 (clone MIH4) coupled to PE (mouse-IgG1 isotype; eBioscience); and anti-human annexin V coupled to FITC (mouse-IgG1 isotype; Beckman Coulter).

Techniques: Expressing

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

Article Title: Armored Inducible Expression of IL-12 Enhances Antitumor Activity of Glypican-3-Targeted Chimeric Antigen Receptor-Engineered T Cells in Hepatocellular Carcinoma.

doi: 10.4049/jimmunol.1800033

Figure Lengend Snippet: FIGURE 6. GPC3-m28Z-mNFAT-mIL-12 T cells improved antitumor immunity against established murine HCC tumor. (A) Left panel, Experimental scheme of in vivo immunocompetent model. Murine T cells at different doses were transferred i.v. into female C57BL/6 mice 8 d after tumor cell in- oculation (n = 6). Right panel, Tumor growth curve for Hepa1-6-GPC3 xenografts treated with indicated murine T cells. Arrow indicates murine T cells infusion. (B) The tumor weight was measured at the study end point. (C) Body weight of each group was measured every 3–4 d (baseline = 100%). (D) The amounts of mIFN-g and mIL-12 (p70) in the sera of treated mice 8 d after therapy were assessed by ELISA. Data shown are the mean 6 SEM. (E) CAR copy numbers in genomic DNA of residual tumors 8 d after therapy were measured by real-time PCR. (F) The sections of formalin-fixed, paraffin- embedded tumor tissue were immunostained with anti-mouse CD8a Ab or anti-mouse Foxp3 Ab. The images were obtained under original magnification 3400. Data shown are from three independent experiments. *p , 0.05, **p , 0.01, ***p , 0.001.

Article Snippet: To detect murine CD8+ T cells and regulatory T cells (Tregs), the sections of formalin-fixed, paraffin-embedded tumor tissue were immunostained with anti-mouse CD8a Ab (Cell Signaling Technology) or anti-mouse Foxp3 mAb (eBioscience), followed by goat anti-rat IgG-HRP (Santa Cruz Biotechnology) or HRP-conjugated rabbit anti-mouse IgG (H&L) (KangChen Biotech).

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation

Article Title: Functional divergence of antigen-specific T-lymphocyte responses in syngeneic graft-versus-host disease.

doi: 10.1016/j.bbmt.2004.05.003

Figure Lengend Snippet: Figure 2. Flow cytometric selection of CLIP antigen-specific T-cell subsets. Splenic T lymphocytes harvested on day 14 after cessation of cyclosporine treatment were subjected to 3-color immunofluorescence. The cells were stained with the biotinylated soluble MHC class II immunoglobulin construct loaded with the N- or C-terminal CLIP variants (counterstained with CyChrome/streptavidin) and with fluorescein isothiocyanate (FITC)–and phycoerythrin (PE)–conjugated antibodies to CD8 and CD25, respectively. The percentage of cells staining with the construct is shown in the histogram; the dot plot displays CD8 and CD25 expression of the cells reacting with the peptide-loaded construct.

Article Snippet: The shaded area, however, repr etween N- and C-terminal flanking domain dependence. eric molecule loaded with unrelated peptides was [ B & M T neffective in isolating CLIP-specific T cells [23]. low cytometric multicolor analysis was used to isoate the antigen-specific T cells and to confirm expresion of the CD4, CD8, and CD25 cell-surface markrs (phycoerythrin- or fluorescein isothiocyanate– onjugated mouse anti-rat CD8, anti-CD4, and CD25 onoclonal antibodies; Serotec, Harlan Bioproducts or Science, Indianapolis, IN).

Techniques: Selection, Staining, Construct, Expressing

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

Article Title: Glucocorticoids regulate TCR-induced elevation of CD4: functional implications.

doi: 10.4049/jimmunol.164.12.6213

Figure Lengend Snippet: FIGURE 1. CD4 expression increases after T cell activation indepen- dently of the stimulus used to activate CD41 T cells. A, Cells were acti- vated with either plate-bound anti-TCR or anti-CD3 mAb (1 mg/well) or the soluble stimulants SEA (1 mg/ml) and Con A (1.5 mg/ml). Cells were cultured for 24–120 h and analyzed by double-immunofluorescence stain- ing with PE-conjugated anti-CD25 mAb and FITC-conjugated anti-CD4 mAb. Unrelated PE- or FITC-conjugated IgG1 mAb were used as controls. Results (representative of four to six independent experiments) are ex- pressed as the mean fluorescence intensity (MFI). Mean CD4 fluorescence of freshly isolated cells was 580 vs 9 for mAb control. Similar results were obtained after staining with OX35 (data not shown). B, FACS plot of SEA-activated CD41 T cells (as shown in A) after 24 h (solid lines) or 96 h (dashed lines) days of incubation. The histograms on the left show control staining with unrelated FITC-conjugated IgG1 mAb. C, CD4 expression by CD251 vs CD252, SEA-treated CD41 T cells (as described in A) after 24–72 h of culture.

Article Snippet: For flow cytometry, FITC-conjugated mouse anti-rat TCRab IgG1 mAb, FITC-conjugated mouse anti-rat CD4 IgG1 mAb (clone W3/25, which recognizes an epitope of domain 1 of CD4), FITCconjugated mouse anti-rat CD8a IgG1 mAb (clone OX8), PE-conjugated anti-CD4 mAb (W3/25), PE-conjugated anti-CD8a mAb (OX8), and PEconjugated mouse anti-rat CD25 IgG1 mAb (clone OX39) were purchased from Serotec.

Techniques: Expressing, Activation Assay, Cell Culture, Staining, Isolation, Control, Incubation

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

Article Title: Glucocorticoids regulate TCR-induced elevation of CD4: functional implications.

doi: 10.4049/jimmunol.164.12.6213

Figure Lengend Snippet: FIGURE 6. Activation-induced TCR down-regulation is not affected by CORT. A, Cells were activated as described in Fig. 1 and cultured for 24–120 h. TCR expression was analyzed by double-immunofluorescence staining with PE-conjugated anti-CD4 mAb and FITC-conjugated anti- TCR mAb. Unrelated PE- or FITC-conjugated IgG1 mAb were used as controls. Results (representative of four independent experiments) are ex- pressed as the percent MFI of TCR expression by fresh cells (5100%; MFI 5 389.8; MFI for mAb control, 8). B, Cells were incubated in the absence or the presence of CORT (1026 M) and were simultaneously ac- tivated with plate-bound anti-TCR mAb (1 mg/well). Cell culture and TCR analysis were performed as described in A.

Article Snippet: For flow cytometry, FITC-conjugated mouse anti-rat TCRab IgG1 mAb, FITC-conjugated mouse anti-rat CD4 IgG1 mAb (clone W3/25, which recognizes an epitope of domain 1 of CD4), FITCconjugated mouse anti-rat CD8a IgG1 mAb (clone OX8), PE-conjugated anti-CD4 mAb (W3/25), PE-conjugated anti-CD8a mAb (OX8), and PEconjugated mouse anti-rat CD25 IgG1 mAb (clone OX39) were purchased from Serotec.

Techniques: Activation Assay, Cell Culture, Expressing, Staining, Control, Incubation