Extended Data Fig. 8: Gut inflammation accelerates TCR^7B8-mediated CNS inflammation.

(A-G) Integrin β7-deficient TCR^7B8 CD4 T cells fail to accumulate in the gut, resulting in less inflammation in both the gut and CNS. Comparison between Itgb7 WT and Itgb7-KO TCR^7B8 CD4 T cells detected in mix-transferred Rag2^−/− hosts. Total CD4 T cell numbers detected in the brain, spinal cord, ileum, colon, and spleen. (A) Schematic representation of Itgb7-sufficient and -deficient naïve TCR^7B8 CD4 T cell transfer to Rag2^−/− hosts. (B) Represented FACS plots of the detection of two different donor CD4 T cells (Itgb7 ^{+/+ or + /−} as CD45.1/CD45.2, and Itgb7^−/− as CD45.2/CD45.2) (Top). Frequency of CD4 T cells in the indicated tissues (Bottom) (n = 7). (C) Total CD4 T cell numbers detected in the spinal cord. (D) Representative FACS plots of IFNγ-, IL-17A-, and TNFα-producing Itgb7 WT and KO TCR^7B8 CD4 T cells detected in the spinal cord. (E-G) Cell numbers of Itgb7 WT and KO TCR^7B8 CD4 T cells producing IFNγ/IL-17A in the spinal cord (E), TNFα in the spinal cord (F), and IL-17A in the Spinal cord (G). Independent experiments were conducted twice using multiple littermate controls (Itgb7 WT TCR^7B8 transferred (n = 13) and Itgb7 KO TCR^7B8 transferred (n = 12) (C, E, F, G). (H) Transferring dysregulated TCR^7B8 CD4 T cells collected from inflamed colon of TCR^7B8 Rag2^−/− mice into the GF WT hosts. (H) Schematic representation of dysregulated TCR^7B8 T cell transfer. CD4 T cells were sorted from the inflamed colons of TCR^7B8 Rag2^−/− mice. The sorted 1 × 10⁶ CD4 T cells were incubated with 300 μg/ml of Anti-CTLA4 and Anti-PD-1 antibodies or control antibodies for 30 min at room temperature and then R.O. transferred into the GF WT recipients and maintained for 19 days (left). ICB were weekly i.p. injected (300 μg of Anti-CTLA4 and Anti-PD-1 antibodies or control antibodies). Their neurological scores were monitored every day following the transfer (right). (I-J) In vitro polarized TCRTg Th17 cell transfer. Sorted naïve CD4 T cells from TCRTg mice were pathogenically or homeostatically polarized in vitro with IL-6/TGFβ/IL-23/IL1β for pathogenic and IL-6/TGFβ/Anti-IFNγ ab/Anti-IL-4 ab. (K) Neurological score (not EAE score) of immunized SPF WT mice with MOG peptide or SFB 7B8 peptide. Two independent experiments were combined. Results are presented as the mean ± SEM, with dots representing individual mice. (B, C, E, F, G) Two-Tailed-Student’s t-test, (H, I, J, K) Two-way ANOVA.

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