Fig. 2.

HAMSB diet does not alter the T_H17 response in CIA mice

(a–c) IFN-γ⁺ and IL-17A⁺ producing CD4⁺T cells in the hind knee joints of CIA mice fed the control HAMS or HAMSB diet two weeks after the initial immunization. Cells were restimulated with PMA and ionomycin for 6 h before staining. Representative flow cytometry contour plots of intracellular IFN-γ and IL-17A staining within CD4⁺TCRβ⁺ gate (a), and the frequency (b) and total number (c) of IL-17A⁺, IFN-γ⁺, and IFN-γ⁺IL-17A⁺ cells (n= 4).

(d–f) RORγt⁺ and IL-17A⁺ cells in the draining lymph nodes (DLNs) of CIA mice fed the control HAMS or HAMSB diet two weeks after the initial immunization. Cells were restimulated with PMA and ionomycin for 6 h. Representative flow cytometry contour plots of intracellular RORγt and IL-17A staining within CD4⁺TCRβ⁺ gate (d), and the frequency (e) and total number (f) of RORγt⁺IL-17A⁺ and RORγt⁻IL-17A⁺ cells (n= 4).

(g, h) IL-17A concentration in the hind knee joint extract (g, n= 6) and serum (h, n= 12) of CIA mice fed the control HAMS or HAMSB diet three weeks after the booster immunization.

(i, j) The frequency in CD4⁺T cells (i) and total number (j) of Nrp-1⁻Foxp3⁺ pT_REG and Nrp-1⁺Foxp3⁺ tT_REG cells in the spleen (SP), DLNs and colonic patches (CoPs) of CIA mice fed control HAMS or HAMSB diet two weeks after the initial immunization (n= 10). Gating strategy is depicted in Figure S4.

Results show one representative experiment of at least two experiments. (Welch's t-test or unpaired two-tailed Student's t-test).