Extended Data Fig. 4: CAR promotes effector T cell persistence in the presence of small intestinal bile acids.
From: CAR directs T cell adaptation to bile acids in the small intestine
a, Percentages of live CD44hi wild-type (B6; CD45.1+; blue) or CAR-deficient (B6.Nr1i3−/−; CD45.1−; red) Teff cells, determined by flow cytometry and gated as in Extended Data Fig. 6a, in tissues from reconstituted B6.Rag1−/− mice over time. Numbers indicate percentages; representative of five mice per tissue and time point. b, Fitness, defined as mean ± s.e.m. log2[fold change (F.C.)] of CAR-deficient (B6.Nr1i3−/−) versus wild-type (B6) Teff cell percentages (n = 5) in tissues from congenically co-transferred Rag1−/− mice over time, determined by flow cytometry as in a. c, Percentage of wild-type (B6, CD45.1+; blue) and CAR-deficient (B6.Nr1i3−/−, CD45.1−; red) naive (CD62Lhi) CD4+ T cells after sorting and mixing, and before in vivo transfer into Rag1−/− mice (input Tnaive); representative of three mixtures used for analysing resulting Teff cells at 2, 4 or 6 weeks after transfer. d, Equal numbers of CD45.1 wild-type (B6; blue) and CD45.2 CAR-deficient (B6.Nr1i3−/−; red) naive CD4+ T cells were transferred together into co-housed Rag1−/− mice with or without the ileal bile acid reuptake transporter ASBT (gene symbol Slc10a2). The resulting Teff cells from siLP were analysed two weeks after T cell transfer via flow cytometry. e, Percentages of live CD44hi wild-type (B6; CD45.1+; blue) or CAR-deficient (B6.Nr1i3−/−; CD45.1−; red) Teff cells, determined by flow cytometry and gated as in Extended Data Fig. 6a, in siLP from week-2 reconstituted B6.Rag1−/− mice. Numbers indicate percentages; representative of 8–10 mice analysed over two independent experiments. f, Mean ± s.e.m. absolute numbers of live CD45.1 wild-type (B6; left) or CD45.2 CAR-deficient (B6.Nr1i3−/−; right) Teff cells, determined by ex vivo flow cytometry as in e, from siLP two weeks after mixed T cell transfer into control (Slc10a2+/+; blue; n = 8) or ASBT-deficient (Slc10a2−/−; red; n = 10) Rag1−/− recipients. Fold changes in cell numbers recovered from ASBT-deficient versus control recipients, as well as P values (two-tailed unpaired Student’s t-test), are indicated.
