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  • 钱莉,董改琴,吴梦芸,荣于馨,陈文艳,刘阳,叶枫,刘露.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[J].第二军医大学学报,2018,39(2):182-187    [点击复制]
  • QIAN Li,DONG Gai-qin,WU Meng-yun,RONG Yu-xin,CHEN Wen-yan,LIU Yang,YE Feng,LIU Lu.Immune complex inhibits Toll-like receptor 9-activated JNK and p38 pathways in B lymphocytes[J].Acad J Sec Mil Med Univ,2018,39(2):182-187   [点击复制]
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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
钱莉1*,董改琴2,吴梦芸1,荣于馨1,陈文艳1,刘阳1,叶枫1,刘露1
0
(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
*通信作者)
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CpG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CpG 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(FcγRb)缺陷小鼠脾脏B淋巴细胞,体外用CpG 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(SP600125,50 μmol/L)和p38抑制剂(SB203580,20 mg/L)处理后,流式细胞术检测CpG 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CpG 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CpG ODN诱导的JNK和p38磷酸化水平,但不能抑制FcγRb缺陷小鼠B淋巴细胞JNK和p38的磷酸化水平。SP600125和SB203580处理后,CpG 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CpG ODN诱导的CD40和CD80表达。
关键词:  免疫复合物  Toll样受体  Fc片段  B淋巴细胞  分化群
DOI:10.16781/j.0258-879x.2018.02.0182
投稿时间:2017-08-24修订日期:2017-10-17
基金项目: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.
Immune complex inhibits Toll-like receptor 9-activated JNK and p38 pathways in B lymphocytes
QIAN Li1*,DONG Gai-qin2,WU Meng-yun1,RONG Yu-xin1,CHEN Wen-yan1,LIU Yang1,YE Feng1,LIU Lu1
(1. Institute of Translational Medcine, Medical College of Yangzhou University, Yangzhou 225001, Jiangsu, China;
2. Department of Pediatrics, Affiliated Hospital of Yangzhou University, Yangzhou 225001, Jiangsu, China
*Corresponding author)
Abstract:
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 CpG oligodeoxynucleotide (ODN) in B lymphocytes. Methods The mice were intraperitoneally injected with CpG ODN or IC plus CpG 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 (FcγRb) knockout mice by MACS. After the isolated cells were stimulated with CpG ODN or IC plus CpG ODN in vitro, the phosphorylation levels of related protein kinases were detected in the B lymphocytes by Western blotting. Following CpG 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 CpG ODN-activated B lymphocytes were detected by flow cytometry. Results IC inhibited CD40 and CD80 expressions on the CpG ODN-activated B lymphocytes in vivo (both P<0.05). IC inhibited the phosphorylation levels of JNK and p38 induced by CpG ODN in B lymphocytes, but did not inhibit them in the B lymphocytes from FcγRb-/- mice. The CD40 and CD80 expressions on the CpG ODN-activated B lymphocytes were significantly decreased after treated with SP600125 and SB203580 (both P<0.01). Conclusion IC can inhibit the CD40 and CD80 expressions induced by TLR9 agonist CpG ODN through inhibiting the JNK and p38 pathways in B lymphocytes.
Key words:  immune complex  Toll-like receptor 9  Fc fragment  B lymphocyte  cluster of differentiation