Fig. 6: Anti-PD-1 treatment increases T cell plaque infiltration and IFNγ production by aortic T cells.
a–k,o–u, IfngYFP/YFPApoe−/− 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. 6 (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 IfngYFP/YFPApoe−/− 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.
