Fig. 5.

Zoledronate (ZOL)-stimulated γδ T cells obtain APC functions via the HSP90-MyD88 pathway. A Resting γδ T cells were treated with 1 μM ZOL for indicated days. The expression level of MyD88 was detected by western blot. B–E Resting γδ T cells were treated with ZOL or ZOL plus MyD88 inhibitor T6167923 (ZOL + T61) for 3 days. B, C The expression levels of MHC molecules on γδ T cells were measured using flow cytometry. D, E The expression levels of CD86 on γδ T cells were measured using flow cytometry. F, G Resting γδ T cells were treated with ZOL or ZOL + T61 for 7 days before MAGEA3 incubation. Then, the γδ T cells were washed and co-cultured with CFSE-labeled CD8⁺ T cells for 10 days at an APC/responder cell ratio of 1:10. Proliferating CFSE⁻ CD8⁺ T cells were measured by flow cytometry. H–K Resting γδ T cells were treated with ZOL or ZOL + T61 for 7 days before MAGEA3 incubation. Then, the γδ T cells were washed and co-cultured with pan T cells for 7 days at an APC/responder cell ratio of 1:10. After that, the cells in each group received the same stimulation as the previous one for 4 h followed by further analysis. H, I Perforin expression levels in CD8⁺ T cells were measured by flow cytometry. J, K IFN-γ expression levels in CD8⁺ T cells were measured by flow cytometry. L Resting γδ T cells were treated, respectively, with ZOL or ZOL + LUM for 7 days. The expression level of MyD88 was detected by western blot. M Resting γδ T cells were treated, respectively, with ZOL or ZOL + T61 for 7 days. The expression levels of HSP90α and HSP90β were detected by western blot. Representative data were shown from 2 to 3 independent experiments. All the values were presented as mean ± SD. *p < 0.05, ***p < 0.001 versus Control, ##p < 0.01, ###p < 0.001. NS, not significant vs. Control