Wang Cong-Cong, Zhang Min, Li Heng, Li Xiao-Li, Yue Long-Tao, Zhang Peng, Liu Ru-Tao, Chen Hui, Li Yan-Bin, Duan Rui-Sheng
Department of Neurology, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan 250014, PR China.
Central Laboratory, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan 250014, PR China.
Neurosci Lett. 2017 Aug 24;656:169-176. doi: 10.1016/j.neulet.2017.05.040. Epub 2017 Jul 22.
We have previously demonstrated that Cysteinyl aspartate-specific proteinase-1 (caspase-1) inhibitor ameliorates experimental autoimmune myasthenia gravis (EAMG) by inhibited cellular immune response, via suppressing DC IL-1 β, CD4 T and γdT cells IL-17 pathways. In this study, we investigated the effect of caspase-1 inhibitor on humoral immune response of EAMG and further explore the underlying mechanisms. An animal model of MG was induced by region 97-116 of the rat AChR α subunit (R97-116 peptide) in Lewis rats. Rats were treated with caspase-1 inhibitor Ac-YVAD-cmk intraperitoneally (i.p.) every second day from day 13 after the first immunization. Flow cytometry, western blot, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) were performed to evaluate the neuroprotective effect of caspase-1 inhibitor on humoral immune response of EAMG. The results showed that caspase-1 inhibitor reduced the relative affinity of anti-R97-116 IgG, suppressed germinal center response, decreased follicular helper T cells, and increased follicular regulatory T cells and regulatory B cells. In addition, we found that caspase-1 inhibitor inhibited humoral immunity response in EAMG rats via suppressing IL-6-STAT3-Bcl-6 pathways. These results suggest that caspase-1 inhibitor ameliorates EAMG by regulating humoral immune response, thus providing new insights into the development of myasthenia gravis and other autoimmune diseases therapies.
我们之前已经证明，半胱氨酸天冬氨酸特异性蛋白酶-1（caspase-1）抑制剂通过抑制细胞免疫反应，经由抑制树突状细胞（DC）白细胞介素-1β、CD4 T细胞和γδT细胞白细胞介素-17通路，改善实验性自身免疫性重症肌无力（EAMG）。在本研究中，我们研究了caspase-1抑制剂对EAMG体液免疫反应的影响，并进一步探讨其潜在机制。用大鼠乙酰胆碱受体（AChR）α亚基的97-116区域（R97-116肽）在Lewis大鼠中诱导重症肌无力动物模型。从首次免疫后第13天起，每隔一天给大鼠腹腔注射（i.p.）caspase-1抑制剂Ac-YVAD-cmk。进行流式细胞术、蛋白质免疫印迹、免疫荧光和酶联免疫吸附测定（ELISA），以评估caspase-1抑制剂对EAMG体液免疫反应的神经保护作用。结果表明，caspase-1抑制剂降低了抗R97-116 IgG的相对亲和力，抑制生发中心反应，减少滤泡辅助性T细胞，并增加滤泡调节性T细胞和调节性B细胞。此外，我们发现caspase-1抑制剂通过抑制白细胞介素-6-信号转导和转录激活因子3-B细胞淋巴瘤-6（IL-6-STAT3-Bcl-6）通路，抑制EAMG大鼠的体液免疫反应。这些结果表明，caspase-1抑制剂通过调节体液免疫反应改善EAMG，从而为重症肌无力和其他自身免疫性疾病的治疗发展提供了新的见解。
