Effects of Proteasome inhibitor bortezomib on apoptosis of murine alloreactive T cells in vitro

Di-san junyi daxue xuebao

YANG Jiacai¹  HUANG Chibing²  YANG Qishun²  WANG Xu²  SONG Yajun² 

[1] State Key Laboratory of Trauma, Burn and Combined Injury, Institute of Burn Research, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038, China;Department of Urology, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037;

关键词: bortezmoib;    t cells;    proliferation;    cell cycle;    apoptosis;    cytokines;   

DOI  :  10.16016/j.1000-5404.201905204

来源: DOAJ

【 摘 要 】

Objective To observe the effects of the proteasome inhibitor bortezomib (BTZ) on the apoptosis of murine alloreactive T cells in vitro. Methods In a one-way mixed lymphocyte culture system, the T lymphocytes from C57BL/6 mice were co-cultured with the splenocytes of BALB/c mice for 24 h. After treatment of the cells with different concentrations of bortezomib for 12 h, the changes in the cell proliferation were assessed using CCK-8 assay. The responses of the co-cultured cells to different culture times and a 12 h treatment with 0.01 μmol/L bortezomib were examined by detecting the changes in the cell apoptosis, cell cycle and expression of the activation-associated markers CD25 and CD69 using flow cytometry; the expression of cleaved caspase-3 and cyclin D1 was detected using Western blotting, and the levels of interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA). Results Compared with the control cells, the bortezomib-treated T cells showed an obviously decreased proliferative activity. Treatment with 0.01 μmol/L bortezomib significantly increased the percentage of apoptotic T cells and enhanced the expression of cleaved caspase-3 in positive correlation with the co-culture time prior to bortezomib treatment (P < 0.05). The proportion of T cells at G1 phase increased and the expression of cyclin D1 decreased significantly in bortezomib-treated cells (P < 0.05), which also showed down-regulated expressions of CD25 and CD69 and lowered levels of IL-4, TNF-α and IFN-γ in the supernatant (P < 0.05). Conclusion Bortezomib can inhibit the proliferation of murine alloreactive T cells in vitro, induce apoptosis and reduce cytokine production in the activated T cells.

【 授权许可】

Unknown