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. 2022 Feb 18;10(4):437–452. doi: 10.1158/2326-6066.CIR-20-0973

Figure 4.

Figure 4. IFNγ hypersecretion by Cbl-b KO CD8+ T cells induces Treg resistance. A, Cytokine secretion profile of activated WT and Cbl-b KO CD8+ T cells. Supernatants were collected on day 3 after stimulation, and LEGENDplex cytokine array was performed (n = 3). B, IFNγ secretion by WT and Cbl-b KO T cells in the presence of Tregs. Supernatants from the Treg suppression assay were collected on day 3 after stimulation, and IFNγ ELISA was performed (n = 3). C, Proliferation of WT T cells cocultured with or without Tregs in the presence of IFNγ or anti-IFNγ (n = 3). D, The percentage suppression of WT CD4+ and CD8+ T cells for each stimulatory condition (n = 3). E, Proliferation of Cbl-b KO CD8+ T cells in the presence of Tregs, with or without anti-IFNγ (n = 3). F, The percentage suppression of WT and Cbl-b KO CD8+ T cells stimulated with or without anti-IFNγ (n = 3). G, Treg suppression assay comparing percentage suppression of WT, Cbl-b KO, and Cbl-b/IFNγ double KO CD8+ T cells in response to Tregs (n = 3). Statistical analyses were performed using Student t test (A and F), and ANOVA with Holm–Sidak test (B, D, and G; *, P < 0. 05; **, P < 0.01; ns, not significant).

IFNγ hypersecretion by Cbl-b KO CD8+ T cells induces Treg resistance. A, Cytokine secretion profile of activated WT and Cbl-b KO CD8+ T cells. Supernatants were collected on day 3 after stimulation, and LEGENDplex cytokine array was performed (n = 3). B, IFNγ secretion by WT and Cbl-b KO T cells in the presence of Tregs. Supernatants from the Treg suppression assay were collected on day 3 after stimulation, and IFNγ ELISA was performed (n = 3). C, Proliferation of WT T cells cocultured with or without Tregs in the presence of IFNγ or anti-IFNγ (n = 3). D, The percentage suppression of WT CD4+ and CD8+ T cells for each stimulatory condition (n = 3). E, Proliferation of Cbl-b KO CD8+ T cells in the presence of Tregs, with or without anti-IFNγ (n = 3). F, The percentage suppression of WT and Cbl-b KO CD8+ T cells stimulated with or without anti-IFNγ (n = 3). G, Treg suppression assay comparing percentage suppression of WT, Cbl-b KO, and Cbl-b/IFNγ double KO CD8+ T cells in response to Tregs (n = 3). Statistical analyses were performed using Student t test (A and F), and ANOVA with Holm–Sidak test (B, D, and G; *, P < 0. 05; **, P < 0.01; ns, not significant).