Journal: Nature Cardiovascular Research
Article Title: Progenitor exhausted PD-1 + T cells are cellular targets of immune checkpoint inhibition in atherosclerosis
doi: 10.1038/s44161-025-00713-2
Figure Lengend Snippet: a – k , o – u , Ifng YFP/YFP Apoe − / − mice were fed an HCD for 10 weeks before administering biweekly intraperitoneal injections of a murine blocking anti-PD-1 antibody or isotype IgG control ( n = 12 per group). Mice received a total of 12 injections over 6 weeks and were euthanized after a total of 16 weeks of diet. b , Quantification of average plaque area of aortic subvalvular cross-sections. c , d , Quantification and representative immunohistochemical staining of T cells (CD3 + ) in aortic root plaques (**** P = 5 × 10 −5 ). e – i , Representative flow cytometry plots and quantification of frequency and numbers of PD-1 subsets of aortic memory CD4 (*** P = 0.0001) and CD8 ( h **** P = 7.4 × 10 −7 , i **** P = 5.2 × 10 −6 ) T cells. j , k , Flow cytometric analysis of frequency of Tim3 + PD-1 + memory CD4 and CD8 aortic T cells (** P = 0.0024) and frequency of IFNγ production of memory CD4 and CD8 aortic T cells (** P = 0.0023, 0.0022). l , m , Representative immunohistochemical images of and quantification of presence of at least one T cell foci in the plaque and/or surrounding adventitia (* P = 0.024). For study details of 3-week anti-PD-1 cohort, see Extended Data Fig. (ctrl IgG n = 12, anti-PD-1 n = 13). n , Impact of presence of tumor (MC38 tumor) during anti-PD-1 therapy ( n = 7 per group) on T cell (CD3 + ) infiltration into subvalvular aortic plaques (** P = 0.0079, * P = 0.026) compared to ctrl IgG ( n = 5 per group), determined by immunohistochemistry. o – q , Quantification and representative immunohistochemical staining of neutrophils (Ly6G + ) in aortic root plaques (** P = 0.0070) and adventitia (**** P = 9.5 × 10 −5 ) in Ifng YFP/YFP Apoe − / − mice treated with PD-1 blockade or isotype IgG ctrl for 6 weeks ( n = 12 per group). r , Quantification of immunohistochemical staining of macrophages (CD68 + ) in aortic root plaques. s , Gene expression analysis of aortic subvalvular cross-sections of ctrl IgG-treated or anti-PD-1-treated mice. Genes are reported as upregulated or downregulated in anti-PD-1-treated mouse compared to isotype IgG control. Adjusted P value is adjusted for FDR. t , u , Concentration of plasma cytokines CXCL9 (** P = 0.0063) and CXCL10 (* P = 0.037). b , c , i , n , p – r , u , Bars denote median, analyzed with two-sided Mann–Whitney U -test. t , Bars denote mean, analyzed with two-sided unpaired t -test. f – h , j , k , Bars denote median, analyzed with two-sided Mann–Whitney U -test or two-sided unpaired t -test. m , Analyzed with one-sided Fisher’s exact test. adj., adjusted; Adv., adventitia; Avg., average; ctrl, control; FC, fold change; i.p., intraperitoneal; w, week.
Article Snippet: Mice were randomly assigned to receive intraperitoneal injections of either murinized and effector-less anti-PD-1 antibody (clone: RMP1-14, anti-mPD-1-mIgG1e3; InvivoGen, mpd1-mab15-50) or isotype control IgG2a (anti-mPD-1-mIgG1e3, InvivoFit; InvivoGen) biweekly for 6 weeks at 10 mg kg −1 ( n = 12 per group).
Techniques: Blocking Assay, Control, Immunohistochemical staining, Staining, Flow Cytometry, Immunohistochemistry, Gene Expression, Concentration Assay, Clinical Proteomics, MANN-WHITNEY