Department of Neurology, Medical University of Lodz, 22 Kopcinskiego str, 90-153 Lodz, Poland.
Cells. 2019 Jul 29;8(8):790. doi: 10.3390/cells8080790.
NK cells (natural killer cells) being a part of the innate immune system have been shown to be involved in immunoregulation of autoimmune diseases. Previously we have shown that HINT1/Hsp70 treatment induced regulatory NK cells ameliorating experimental autoimmune encephalomyelitis (EAE) course and CD4+ T cells proliferation. NK cells were isolated from mice treated with HINT1/Hsp70 and co-cultured with proteolipid protein (PLP)-stimulated CD4+ T cells isolated from EAE mice. Cell proliferation was assessed by thymidine uptake, cytotoxicity by lactate dehydrogenase (LDH) release assay and fluorescence activated cell sorting (FACS) analysis, protein expression by Western blot, mRNA by quantitative RT-PCR. Gene related to anergy in lymphocytes (GRAIL) expression was downregulated by specific siRNA and GRAIL overexpression was induced by pcDNA-GRAIL transfection. HINT1/Hsp70 pretreatment of EAE SJL/J mice ameliorated EAE course, suppressed PLP-induced T cell proliferation by enhancing T cell expression of GRAIL as GRAIL downregulation restored T cell proliferation. HINT1/Hsp70 treatment induced immunoregulatory NK cells which inhibited PLP-stimulated T cell proliferation not depending on T cell necrosis and apoptosis. This immunoregulatory NK cell function depended on NK cell expression of GRAIL as GRAIL downregulation diminished inhibition of NK cell suppression of T cell proliferation. Similarly GRAIL overexpression in NK cells induced their regulatory function. HINT1/Hsp70 treatment generated regulatory NK cells characterized by expression of GRAIL.
自然杀伤细胞（NK 细胞）作为先天免疫系统的一部分，已被证明参与自身免疫性疾病的免疫调节。此前我们已经表明，HINT1/Hsp70 治疗诱导调节性 NK 细胞改善实验性自身免疫性脑脊髓炎（EAE）病程和 CD4+T 细胞增殖。从接受 HINT1/Hsp70 治疗的小鼠中分离 NK 细胞，并与从 EAE 小鼠中分离的蛋白脂质蛋白（PLP）刺激的 CD4+T 细胞共培养。通过胸苷摄取评估细胞增殖，通过乳酸脱氢酶（LDH）释放测定和荧光激活细胞分选（FACS）分析评估细胞毒性，通过 Western blot 评估蛋白质表达，通过定量 RT-PCR 评估 mRNA。通过特异性 siRNA 下调与淋巴细胞无反应性相关的基因（GRAIL）的表达，并通过 pcDNA-GRAIL 转染诱导 GRAIL 过表达。HINT1/Hsp70 预处理 SJL/J 型 EAE 小鼠可改善 EAE 病程，通过增强 T 细胞 GRAIL 的表达来抑制 PLP 诱导的 T 细胞增殖，因为 GRAIL 下调恢复了 T 细胞增殖。HINT1/Hsp70 治疗诱导的免疫调节性 NK 细胞抑制 PLP 刺激的 T 细胞增殖，而不依赖于 T 细胞坏死和凋亡。这种免疫调节性 NK 细胞功能依赖于 NK 细胞 GRAIL 的表达，因为 GRAIL 下调减弱了抑制 NK 细胞抑制 T 细胞增殖的作用。同样，NK 细胞中 GRAIL 的过表达诱导其调节功能。HINT1/Hsp70 治疗产生了具有 GRAIL 表达特征的调节性 NK 细胞。
Zotero 插件
