Journal: bioRxiv

Article Title: PD-L1:CD80 Heterodimer Triggers CD28 While Repressing Both PD-1 and CTLA4 Pathways

doi: 10.1101/615138

Figure Lengend Snippet: (A) Representative flow cytometry histograms of CTLA4-Fc binding to the three indicated types of Raji B cells at the indicated concentrations. The MFI of bound CTLA4-Fc labeled by anti-human IgG AF647 was plotted against [Ctla4-Fc] (means ± SEM, n = 3), and fit with “one site – specific binding” model using Graphpad Prism to obtain K D values. (B) A Jurkat–Raji co-culture assay analyzing how PD-L1 interferes with CTLA4 mediated CD80 depletion. Shown on the left are cartoons depicting the co-cultured cells: CTLA4–mGFP transduced Jurkat (CTLA4+) or wild-type Jurkat (CTLA4-) were co-cultured with either PD-L1–mCherry transduced Raji (CD80+/PD-L1+) or wild-type Raji lacking PD-L1(CD80+/PD-L1-) (see Methods ). Shown in the middle are flow cytometry histograms of CD80 expression (anti-CD80 APC) on Raji cells before (0 min) and after co-culture (30 min) and on the right are representative confocal images for the Jurkat–Raji conjugate from three independent replicates. Scale bars:10 μm. (C) An independent Jurkat–Raji conjugation assay examining how PD-L1 and CTLA4 blocking antibodies affect CD80 levels. Experiments were conducted as in (B) except pretreating the indicated cell type with atezolizumab (blocking PD-L1:CD80 cis -interaction) or ipilimumab (blocking CTLA4:CD80 trans -interaction). Shown in the middle and right are representative flow cytometry histograms of CD80 expression (anti-CD80 APC) and confocal images for the Jurkat–Raji conjugate from three independent replicates. Scale bars: 10 μm. (D) Flow cytometry histograms of CD80 and CD86 surface expressions on splenic DCs co-cultured with Treg cells in the presence or absence of a PD-L1 blockade antibody and/or a CTLA4 blockade antibody for 16 hr. Shown are representative flow cytometry histograms for CD80 and CD86 levels on DCs and bar graphs summarizing the result from three independent replicates. See also .

Article Snippet: For blockade treatment in , both SEE-loaded Raji and Jurkat were treated with 2 μg of either atezolizumab (Selleckchem, catalog A2004) or ipilimumab (Selleckchem, catalog A2001) per million cells for 15 minutes prior to mixing, and blockade antibodies were kept in the coculture until the staining step.

Techniques: Flow Cytometry, Binding Assay, Labeling, Co-culture Assay, Cell Culture, Expressing, Co-Culture Assay, Conjugation Assay, Blocking Assay

Journal: Journal of Hematology & Oncology

Article Title: Improvement of the anticancer efficacy of PD-1/PD-L1 blockade via combination therapy and PD-L1 regulation

doi: 10.1186/s13045-022-01242-2

Figure Lengend Snippet: Completed clinical trials of approved PD-1/PD-L1 inhibitors in combination with other approved treatment strategy

Article Snippet: , , Ipilimumab , CTLA-4 antibody , Malignant pleural mesothelioma , Phase 2 , NCT03048474.

Techniques: Clinical Proteomics, Capsules, Virus, Irradiation

Journal: Frontiers in Immunology

Article Title: Combining Immunotherapy with Oncogene-Targeted Therapy: A New Road for Melanoma Treatment

doi: 10.3389/fimmu.2015.00046

Figure Lengend Snippet: Ongoing clinical studies combining immunotherapy and oncogene-targeted therapy .

Article Snippet: Ipilimumab (Yervoy, Bristol-Myers Squibb) is a monoclonal antibody (IgG1) directed against CTLA-4 that was developed for systemic anti-tumor immunotherapy.

Techniques: Biomarker Discovery, Inhibition, Mutagenesis, Injection, Activity Assay, Immunopeptidomics, Capsules, Vaccines, Derivative Assay, Expressing, Amplification, Ex Vivo