Fig. 4: γδ T cells respond to C. neoformans (Cn) Δsgl1 via the production of IFNγ and IL-17A.

a–d. Splenocytes from uninfected TCRβ^−/− mice were processed and cultured ex vivo with plate-bound anti-TCRγδ. These cultured cells were stimulated with PBS, live C. neoformans WT, C. neoformans Δsgl1, or C. neoformans Δcap59Δsgl1 (a, b) or PBS, heat-killed (HK) C. neoformans WT, HK C. neoformans Δsgl1, or HK C. neoformans Δcap59Δsgl1 (c, d). On days 1, 3, and 5 post stimulation, supernatants were collected and assessed for production of IFNγ (a, c) or IL-17A (b, d) via ELISA. e, f. γδ T cells were purified from the spleens of uninfected C57BL/6 mice via MACS separation kit and cultured ex vivo with (+) or without (−) plate-bound anti-TCRγδ, stimulated with PBS, live C. neoformans WT, C. neoformans Δsgl1, or C. neoformans Δcap59Δsgl1, and assessed for IFNγ (e) and IL-17A (f) production on days 1, 3, and 5 post stimulation. Graphed data represents the mean ± SD and are representative of 2 independent experiments (n = 3 mice/group/timepoint for each biological replicate) (a–d) or a single replicate of n = 3 mice/group/timepoint (e, f). Significance was determined by a two-way ANOVA using Šídák’s multiple comparisons test for P value adjustment, and significance is denoted as *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001.