Figure 7.

Tolerized mice have fewer OVA-responsive T cells than control mice or RGMb mAb–treated mice. B cell–depleted splenocytes prepared on day 19 from mice treated as in Fig. 6 a were labeled with CFSE and cultured with OVA for 4 d. Cells were stained for CD3 and CD4 and subgated on CD3⁺CD4⁺ cells for analysis of CFSE dilution. (a) Flow cytometry plots show data from one representative mouse and (b) shows mean ± SEM of three mice/group. (c) On day 4, cultures were harvested as in panel a and supernatants were collected and assayed for cytokine production. Total IL-4, IL-5, and IFN-γ in culture supernatants is shown. (d) Cytokine per cell was calculated as total cytokine amount in supernatant divided by the number of CFSE low cells. Data are mean ± SEM; **, P < 0.01; ***, P < 0.005; n = 3, Student’s t test. Data are representative of two experiments.