Fig 4.

(a to h) SMEZ-2 M1 fails to stimulate cytokine production or in vivo expansion of Vβ11⁺ T cells. (a) Human PBMCs were incubated with 100 ng/ml SMEZ-2 wild type (wt), 100 ng/ml M1, or no antigen. The cytokine response was quantified in culture supernatants after 72 h at 37°C. Results are shown as the mean concentration ± SD of five donors. (b to d) The serum cytokine response was measured 3 h after intraperitoneal injection with 10 μg SMEZ-2, M1, or PBS in age-matched wild-type C57BL/6 (b), DR3-DQ2 (c), and DR4-DQ8 (d) mice. Data are presented as the mean ± SD for 2 mice per treatment per strain; a total of 18 mice were used per study. These data are representative of two independent experiments. (e to h) Three days after injection of 10 μg SMEZ-2, SMEZ-2 M1, or PBS, T cell populations were enumerated by combining data from whole-spleen-cell counts with flow cytometric analysis: CD4⁺ Vβ8⁺ (e), CD8⁺ Vβ8⁺ (f), CD4⁺ Vβ11⁺ (g), and CD8⁺ Vβ11⁺ (h). The fold change in T cell numbers was determined relative to the PBS control group. Data were combined from two independent experiments (n = 4 per treatment group; n = 36 mice in total). Each point represents a single mouse, and the horizontal bar is the median value. P values were calculated using a one-tailed Mann-Whitney test.