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

Article Title: Helper B cells promote cytotoxic T cell survival and proliferation independently of antigen presentation through CD27/CD70 interactions.

doi: 10.4049/jimmunol.180.3.1362

Figure Lengend Snippet: FIGURE 5. Function of TILs/B cell couplets. A, Representative expression of CD27, CD28, CD70, and CD80 in TILs, and in fresh CD19 cells. B, F.I. of TILs after a 4-day expansion alone or in the presence of fresh heterologous B cells. TILs, or CD19 cells, were blocked at the beginning of the culture with anti-CD70 (dashed bar), and anti-CD80 (diamond bar) mAbs, respectively, or with the proper Ig control (light-gray bar is the control for CD70 block, dark-gray bar for CD80 block) (n 4). C, F.I. of CTL after 9 days culture alone or after cross-linking of human recombinant CD27, anti-CD70 mAb, or a combination of the two. The anti-CD19 mAb used as an isotype matched negative control, whereas the pan T cell stimulator OKT3 mAb was used as positive control. PBMC were expanded in presence of rIL-2 with (CD8Flu, black bars) or without (CD8 alone, white bars) Ag-specific stimulation with the Flu peptide. Student’s t test p values refer to significant differences between OKT-3 stimulated cultures and CD8 alone cultures (* 0.05, ** 0.01); all other conditions did not yield significant differences compared with CD8 alone cultures.

Article Snippet: The specificity of the requirements for CD27 stimulation of T cell proliferation by B cells through CD27 was tested by cross-linking human recombinant CD27 (R&D Systems) 10 ng/ml, anti-human CD70 mAb (Ancell), or antiCD3/OKT3 mAb (Ortho Biotech) to culture plates.

Techniques: Expressing, Control, Blocking Assay, Recombinant, Negative Control, Positive Control

Journal: Clinical Cancer Research

Article Title: Preclinical Characterization of SGN-70, a Humanized Antibody Directed against CD70

doi: 10.1158/1078-0432.ccr-08-0493

Figure Lengend Snippet: Fig. 5. SGN-70 does not bind mouse or rat CD70 but binds nonhuman primate CD70. A, murine and rat splenocytes isolated from immunocompetent animals were treated with anti-IgM + anti-CD40 to activate B cells and induce CD70 expression. To evaluate B cells, activated splenocytes were stained with species specific anti-B220 antibodies and histograms gated on B220+ cells. Left, cells were stained with AF488-conjugated SGN-70. Right, murine cells were stained with biotinylated anti-mouse CD70 (clone FR70) and detected with AF488-conjugated avidin; rat CD70 was detected using a recombinant human CD27-Fc fusion protein followed by a fluorochrome-conjugated anti-Fc reagent. B, left, cynomolgus splenocytes and human peripheral blood mononuclear cells were stimulated with anti-CD3 for 5 d to induce CD70 expression. Cells were stained with anti-CD2 and histograms gated on CD2+ cells. Right, 293 cells were transfected with CD70 cDNA isolated from cynomolgus monkey peripheral blood mononuclear cells activated to express CD70. Cells in B were stained with biotinylated SGN-70 and detected with AF488-conjugated avidin. A and B, CD70 (filled histograms) and isotype control (open histograms).

Article Snippet: A biotinylated goat anti-human CD27 antibody (R&D Systems) was used at 5 Ag/mL and detected using Bond Intense R Detection kit (Leica Biosystems) with 3,3¶-diaminobenzidine as substrate.

Techniques: Isolation, Expressing, Staining, Avidin-Biotin Assay, Recombinant, Transfection, Control