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目的 :评估去乙酰化酶(HDAC)抑制剂丙戊酸(VPA)对小鼠树突状细胞(DC)免疫调节的作用。方法:利用C57BL/6小鼠骨髓前体细胞体外诱导DC生成,评估0.5、1 mmol/L VPA对DC分化、抗原递呈能力及细胞因子释放的调控作用,及DC通过该免疫调节表型在混合淋巴细胞反应中对同种反应性CD8+、CD4+T淋巴细胞增殖分化的影响。结果:0.5、1 mmol/L VPA明显干扰DC分化(P<0.05),0.5、1、2、4 mmol/L VPA处理的成熟DC与磷酸盐缓冲液(PBS)处理的对照组DC其早期及晚期凋亡比例比较差异无统计学意义。经VPA处理后DC抗原递呈能力无明显改变,其促炎因子白介素(IL)-12 p70在基因(1 mmol/L组基因相对表达量峰值620.03±58.82、55.22±5.02,对照组2349.30±346.34、147.26±8.29,P<0.001)及蛋白水平(1 mmol/L组蛋白分泌量26.90±5.22,对照组979.98±57.22,P<0.001)显著下调。混合淋巴细胞反应体系中VPA处理的DC免疫调节效应体现在对同种反应性CD4+T细胞增殖抑制(羟基荧光素二醋酸盐琥珀酰亚胺酯低表达群体为13.15%±6.26%,对照组23.54%±9.81%,P<0.05)及诱导凋亡作用(膜联蛋白Ⅴ16.75%±3.06%,7-氨基放线菌素7.38%±1.88%,对照组为4.33%±2.14%、1.16%±0.88%,P<0.01及P<0.05),而对初始CD4+T淋巴细胞向各亚群的分化无显著影响。与经VPA处理DC共培养的CD8+T细胞其细胞毒作用轻度受抑。结论:0.5、1 mmol/L VPA可诱导小鼠DC产生低表达促炎因子IL-12,可诱导同种反应性CD4+T淋巴细胞增殖抑制和凋亡增多的免疫耐受表型。
AIM: To assess the effect of valproate (VPA), a deacetylase (HDAC) inhibitor, on the immunoregulation of mouse dendritic cells (DCs). Methods: DCs were induced by C57BL / 6 mouse bone marrow progenitor cells in vitro. The effects of 0.5 and 1 mmol / L VPA on DC differentiation, antigen presenting ability and cytokine release were evaluated. The immunoregulatory phenotype Effect of Mixed Lymphocyte Reaction on Proliferation and Differentiation of Alloreactive CD8 + and CD4 + T Lymphocytes. Results: 0.5 and 1 mmol / L VPA significantly interfered with DC differentiation (P <0.05). Compared with DCs treated with 0.5, 1, 2 and 4 mmol / L VPA, mature DCs treated with VPA and phosphate buffered saline (PBS) There was no significant difference in late apoptotic ratio. After VPA treatment, there was no significant change in DC antigen presenting ability. The expression of proinflammatory cytokine interleukin (IL) -12 p70 in gene (1 mmol / L, 620.03 ± 58.82, 55.22 ± 5.02, 2349.30 ± 346.34 , 147.26 ± 8.29, P <0.001) and protein level (1 mmol / L histone secretion 26.90 ± 5.22, control group 979.98 ± 57.22, P <0.001). The immunomodulatory effect of VPA-treated DC in mixed lymphocyte reaction system was shown to be inhibited proliferation of alloreactive CD4 + T cells (13.15% ± 6.26% of the low-expression group of hydroxyfluorescein diacetate succinimidyl ester, control (23.54% ± 9.81%, P <0.05) and induced apoptosis (annexin Ⅴ16.75% ± 3.06%, 7.38% ± 1.88%, control group was 4.33% ± 2.14% 1.16% ± 0.88%, P <0.01 and P <0.05), but had no significant effect on the differentiation of naive CD4 + T lymphocytes to each subpopulation. CD8 + T cells co-cultured with VPA-treated DCs were mildly inhibited. CONCLUSION: 0.5,1 mmol / L VPA can induce DCs to produce low expression proinflammatory cytokine IL-12, which can induce the proliferation of allogeneic CD4 + T lymphocytes and increase the immune tolerance phenotype.