Journal: Journal for immunotherapy of cancer

Article Title: XTX101, a tumor-activated, Fc-enhanced anti-CTLA-4 monoclonal antibody, demonstrates tumor-growth inhibition and tumor-selective pharmacodynamics in mouse models of cancer.

doi: 10.1136/jitc-2023-007785

Figure Lengend Snippet: Figure 4 In vitro function of XTX101. (A,B) In vitro inhibition of human CTLA-4 binding to CD80 (A) and CD86 (B) by XTX100 (black), XTX101 (red), and activated XTX101 (blue), as assessed by ELISA. (C) In vitro activity of intact (red) and activated XTX101 (blue) compared with XTX100 (black) in antibody-dependent cellular cytotoxicity (ADCC) reporter bioassay. (D) IL-2 production of SEB-stimulated human peripheral blood mononuclear cells (PBMCs) incubated with XTX100 (black), XTX101 (red), and activated XTX101 (blue), as measured by ELISA. Fold-change from isotype at highest mAb concentration is reported. ND, not determined.

Article Snippet: Serial dilutions of test articles were added to the washed ELISA plates followed by addition of a 2.6 μg/ mL solution of recombinant human CD80 or CD86 (9050- B1- 100 or 9090- B2- 100, R&D Systems).

Techniques: In Vitro, Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Bioassay, Incubation, Concentration 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 CD28-huFc staining of the indicated types of Raji cells. Bound CD28-huFc was labeled by AF647 anti-human IgG Fc, the MFI of which was plotted against (CD28-huFc. Shown in gray are Raji (CD80+CD86−) cells stained by isolated huFc domain. Means ± SEM, n = 3.

Article Snippet: The SLBs were then overlaid with 200 μL of either 3 nM human PD-1-His (Sino Biological, 10377-H08H), 3 nM human CD80–His (Sino Biological, 10698-H08H), or 3 nM human CD86–His (Sino Biological, 10699-H08H).

Techniques: Flow Cytometry, Staining, Labeling, Isolation

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 TIRF images of PD-L1 LUVs captured by PD-1 SLB, CD80 SLB, or CD86 SLB; each LUV is registered as a green spot. Bar graph summarizes the fluorescence intensity (FI) of the LUV channel under indicated conditions, normalized to the intensity of the condition with PD-1 SLBs. Data are means ± SEM, n = 3. Scale bars, 5 μm.

Article Snippet: The SLBs were then overlaid with 200 μL of either 3 nM human PD-1-His (Sino Biological, 10377-H08H), 3 nM human CD80–His (Sino Biological, 10698-H08H), or 3 nM human CD86–His (Sino Biological, 10699-H08H).

Techniques: Fluorescence

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: The SLBs were then overlaid with 200 μL of either 3 nM human PD-1-His (Sino Biological, 10377-H08H), 3 nM human CD80–His (Sino Biological, 10698-H08H), or 3 nM human CD86–His (Sino Biological, 10699-H08H).

Techniques: Flow Cytometry, Staining, Labeling, Isolation

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) On the left are flow-cytometry histograms of CD80 and CD86 surface levels on DCs and macrophages (Macs) isolated from tumor tissues of 4T1 implanted BALB/C female mice treated with anti-PD-L1 (magenta traces), anti-PD-1 (cyan traces), or control IgG (black traces). On the right are bar graphs summarizing the MFI of CD80 and CD86 staining under the indicated conditions. Data are shown as mean ± SEM, n ≥ 3 mice.

Article Snippet: The SLBs were then overlaid with 200 μL of either 3 nM human PD-1-His (Sino Biological, 10377-H08H), 3 nM human CD80–His (Sino Biological, 10698-H08H), or 3 nM human CD86–His (Sino Biological, 10699-H08H).

Techniques: Flow Cytometry, Isolation, Staining

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: KEY RESOURCES TABLE

Article Snippet: The SLBs were then overlaid with 200 μL of either 3 nM human PD-1-His (Sino Biological, 10377-H08H), 3 nM human CD80–His (Sino Biological, 10698-H08H), or 3 nM human CD86–His (Sino Biological, 10699-H08H).

Techniques: Recombinant, Isolation, Enzyme-linked Immunosorbent Assay, Cell Isolation, Software, Imaging