Journal: Cancer cell

Article Title: Selective FcγR Co-engagement on APCs Modulates the Activity of Therapeutic Antibodies Targeting T Cell Antigens

doi: 10.1016/j.ccell.2018.05.005

Figure Lengend Snippet: (A) BALB/c mice with established CT26 tumors (50–80 mm3) were treated with a single 100-μg intraperitoneal (i.p.) dose of anti-CTLA-4, or mIgG2a isotype control mAb. Individual tumor growth rates (n = 9 mice/group) are shown on the right.

Article Snippet: To collect kinetic binding data, recombinant human CTLA-4 his-tagged (Sino Biological) or Fc-fusion (R&D Systems) was injected over the flow cells at concentrations of 20, 6.7, 2.2, and 0.7 nM at a flow rate of 30 μl/minute and at a temperature of 25°C.

Techniques:

Journal: Cancer cell

Article Title: Selective FcγR Co-engagement on APCs Modulates the Activity of Therapeutic Antibodies Targeting T Cell Antigens

doi: 10.1016/j.ccell.2018.05.005

Figure Lengend Snippet: (A and B) Proliferation (Ki67+) of SEB-specific (Vβ8+, black lines) and non-specific (Vβ2+, gray lines) T cells (A) and the percentage of Vβ8+ and Vβ2+ T cells (B) evaluated by flow cytometry in the peripheral blood pre- (day 0) and post-treatment (days 3, 6, and 10) of C57BL/6 mice administered i.p. with 150 μg of SEB together with a 100-μg dose of anti-CTLA-4 mAb or mIgG2a isotype control mAb (n = 5 mice/group). A two-way ANOVA was used to calculate significance.

Article Snippet: To collect kinetic binding data, recombinant human CTLA-4 his-tagged (Sino Biological) or Fc-fusion (R&D Systems) was injected over the flow cells at concentrations of 20, 6.7, 2.2, and 0.7 nM at a flow rate of 30 μl/minute and at a temperature of 25°C.

Techniques: Flow Cytometry

Journal: Cancer cell

Article Title: Selective FcγR Co-engagement on APCs Modulates the Activity of Therapeutic Antibodies Targeting T Cell Antigens

doi: 10.1016/j.ccell.2018.05.005

Figure Lengend Snippet: (A) C57BL/6 mice were administered i.p. with 150 μg of SEB together with a 100-μg i.p. dose of anti-CTLA-4, or a mIgG2a isotype control mAb (n = 5 mice/group). FoxP3+ Treg cell frequency was evaluated in the peripheral blood by flow cytometry pre- (day 0) and post-treatment (days 3, 6, and 10).

Article Snippet: To collect kinetic binding data, recombinant human CTLA-4 his-tagged (Sino Biological) or Fc-fusion (R&D Systems) was injected over the flow cells at concentrations of 20, 6.7, 2.2, and 0.7 nM at a flow rate of 30 μl/minute and at a temperature of 25°C.

Techniques: Flow Cytometry

Journal: Cancer cell

Article Title: Selective FcγR Co-engagement on APCs Modulates the Activity of Therapeutic Antibodies Targeting T Cell Antigens

doi: 10.1016/j.ccell.2018.05.005

Figure Lengend Snippet: (A) IL-2 production (day 4) by human PBMCs stimulated with 100 ng/mL of SEA peptide together with increasing concentrations of anti-CTLA-4, or isotype control mAbs.

Article Snippet: To collect kinetic binding data, recombinant human CTLA-4 his-tagged (Sino Biological) or Fc-fusion (R&D Systems) was injected over the flow cells at concentrations of 20, 6.7, 2.2, and 0.7 nM at a flow rate of 30 μl/minute and at a temperature of 25°C.

Techniques:

Journal: Cancer cell

Article Title: Selective FcγR Co-engagement on APCs Modulates the Activity of Therapeutic Antibodies Targeting T Cell Antigens

doi: 10.1016/j.ccell.2018.05.005

Figure Lengend Snippet: (A) IL-2 production (day 4) by human PBMCs stimulated with 100 ng/mL of SEA peptide together with anti-CTLA-4 mAb variants or corresponding hIgG1 isotype control mAbs.

Article Snippet: To collect kinetic binding data, recombinant human CTLA-4 his-tagged (Sino Biological) or Fc-fusion (R&D Systems) was injected over the flow cells at concentrations of 20, 6.7, 2.2, and 0.7 nM at a flow rate of 30 μl/minute and at a temperature of 25°C.

Techniques:

Journal: Cancer cell

Article Title: Selective FcγR Co-engagement on APCs Modulates the Activity of Therapeutic Antibodies Targeting T Cell Antigens

doi: 10.1016/j.ccell.2018.05.005

Figure Lengend Snippet: (A) Enforced cell surface expression of CTLA-4 by Jurkat IL-2-luc T cells and endogenous expression of CD80 and CD86 on Raji APCs as compared with isotype control mAbs (gray).

Article Snippet: To collect kinetic binding data, recombinant human CTLA-4 his-tagged (Sino Biological) or Fc-fusion (R&D Systems) was injected over the flow cells at concentrations of 20, 6.7, 2.2, and 0.7 nM at a flow rate of 30 μl/minute and at a temperature of 25°C.

Techniques: Expressing

Journal: Cancer cell

Article Title: Selective FcγR Co-engagement on APCs Modulates the Activity of Therapeutic Antibodies Targeting T Cell Antigens

doi: 10.1016/j.ccell.2018.05.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: To collect kinetic binding data, recombinant human CTLA-4 his-tagged (Sino Biological) or Fc-fusion (R&D Systems) was injected over the flow cells at concentrations of 20, 6.7, 2.2, and 0.7 nM at a flow rate of 30 μl/minute and at a temperature of 25°C.

Techniques: Generated, Recombinant, Staining, Expressing, Software

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) A Jurkat-Raji co-culture assay analyzing how PD-L1 interferes with CTLA-4-mediated CD80 depletion. Cartoons on the left depict the co-cultured cells. On the immediate right are representative flow-cytometry histograms of CD80 expression (anti-CD80 allophycocyanin) on Raji cells before (0 h) and after co-culture (0.5 h). Further on the right are representative confocal images for the Jurkat-Raji conjugate (scale bars, 10 μm). Rightmost is a bar graph showing CD80 MFI of Raji at 0.5 h, normalized to CD80 MFI at 0 h (mean ± SEM, n = 4).

Article Snippet: This home-made SNAP–CTLA-4–His 6 and human CD80–His, purchased from Sino Biological, were subjected to gel filtration chromatography, and monomeric fractions of each collected and used in the SPR assays.

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

Journal: Immunity

Article Title: PD-L1:CD80 Cis -Heterodimer Triggers the Co-stimulatory Receptor CD28 While Repressing the Inhibitory PD-1 and CTLA-4 Pathways

doi: 10.1016/j.immuni.2019.11.003

Figure Lengend Snippet: (A) Representative flow-cytometry histograms of CTLA-4-huFc staining of the indicated types of Raji cells. Bound CTLA-4-huFc was labeled by AF647 anti-human IgG Fc, the MFI of which was plotted against (CTLA-4-huFc). Shown in gray are Raji (CD80+CD86−) cells stained by isolated huFc domain. Means ± SEM, n ≥ 3.

Article Snippet: This home-made SNAP–CTLA-4–His 6 and human CD80–His, purchased from Sino Biological, were subjected to gel filtration chromatography, and monomeric fractions of each collected and used in the SPR assays.

Techniques: Flow Cytometry, Staining, Labeling, Isolation