免疫复合物抑制B淋巴细胞内Toll样受体9激活的JNK和p38通路  

Immune complex inhibits Toll-like receptor 9-activated JNK and p38 pathways in B lymphocytes

作　　者：钱莉 董改琴[2] 吴梦芸 荣于馨 陈文艳 刘阳 叶枫 刘露 

机构地区：[1]扬州大学医学院转化医学研究院,扬州225001 [2]扬州大学附属医院儿科,扬州225001

出　　处：《第二军医大学学报》2018年第2期182-187,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81001308;81373130;81771689);江苏省自然科学基金(BK2010315);扬州大学中青年学术带头人资助项~~

摘　　要：目的考察B淋巴细胞内免疫复合物(IC)对Toll样受体9(TLR9)激动剂Cp G寡脱氧核苷酸(ODN)诱导的CD40和CD80高表达信号通路的抑制作用。方法给小鼠腹腔注射Cp G ODN和IC后,用免疫磁珠法分选小鼠脾脏CD19+B淋巴细胞,流式细胞术检测B淋巴细胞表面CD40和CD80的表达。免疫磁珠法分选野生型和免疫球蛋白G Fcγ段受体Ⅱb(FcγRⅡb)缺陷小鼠脾脏B淋巴细胞,体外用Cp G ODN和(或)IC刺激后,蛋白质印迹法检测细胞内相关蛋白激酶的磷酸化水平。用JNK抑制剂(SP600125,50μmol/L)和p38抑制剂(SB203580,20 mg/L)处理后,流式细胞术检测Cp G ODN活化B淋巴细胞表面CD40和CD80的表达。结果体内实验结果显示,IC抑制Cp G ODN活化B淋巴细胞表面CD40和CD80的表达(P均<0.05)。IC抑制B淋巴细胞内Cp G ODN诱导的JNK和p38磷酸化水平,但不能抑制FcγRⅡb缺陷小鼠B淋巴细胞JNK和p38的磷酸化水平。SP600125和SB203580处理后,Cp G ODN活化B淋巴细胞表面CD40和CD80的表达均下调(P均<0.01)。结论 B淋巴细胞内IC通过抑制JNK和p38通路抑制TLR9激动剂Cp G ODN诱导的CD40和CD80表达。Objective To explore the inhibitory effect of immune complex（IC） on the signal pathways of high-expressed CD40 and CD80 induced by Toll-like receptor（TLR9） agonist Cp G oligodeoxynucleotide（ODN） in B lymphocytes. Methods The mice were intraperitoneally injected with Cp G ODN or IC plus Cp G ODN, and the spleen CD19＋ B lymphocytes were sorted by magnetic-activated cell sorting（MACS）. The expressions of CD40 and CD80 on the B lymphocytes were detected by flow cytometry. The spleen B lymphocytes were isolated from wild type and immunoglobulin G Fcγ receptor Ⅱb（FcγRⅡb） knockout mice by MACS. After the isolated cells were stimulated with Cp G ODN or IC plus Cp G ODN in vitro, the phosphorylation levels of related protein kinases were detected in the B lymphocytes by Western blotting. Following Cp G ODN stimulation, the B lymphocytes were treated with JNK p38 inhibitor SP600125（50 μmol/L） or p38 inhibitor SB203580（20 mg/L）, and then the CD40 and CD80 expression levels on the Cp G ODN-activated B lymphocytes were detected by flow cytometry. Results IC inhibited CD40 and CD80 expressions on the Cp G ODN-activated B lymphocytes in vivo（both P0.05）. IC inhibited the phosphorylation levels of JNK and p38 induced by Cp G ODN in B lymphocytes, but did not inhibit them in the B lymphocytes from FcγRⅡb-/-mice. The CD40 and CD80 expressions on the Cp G ODN-activated B lymphocytes were significantly decreased after treated with SP600125 and SB203580（both P0.01）. Conclusion IC can inhibit the CD40 and CD80 expressions induced by TLR9 agonist Cp G ODN through inhibiting the JNK and p38 pathways in B lymphocytes.

关 键 词：免疫复合物 TOLL样受体 Fc片段 B淋巴细胞 分化群 

分 类 号：R392.1[医药卫生—免疫学]