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作 者:秦宏超[1] 芮程磊 陈文艳[1] 傅奕[1] 贾筱琴[1] 田芳[1] 龚卫娟[1] 季明春[1] 钱莉[1]
出 处:《扬州大学学报(农业与生命科学版)》2016年第2期17-21,共5页Journal of Yangzhou University:Agricultural and Life Science Edition
基 金:国家自然科学基金资助项目(81001308;81373130;81172785;81273214;81101265);国家级大学生创新创业训练计划项目(2013-12);扬州大学"新世纪人才工程"项目(2012-12)
摘 要:利用免疫磁珠法分选小鼠脾脏CD19+B细胞,体外用CpG寡脱氧核苷酸(ODN)刺激24h后,流式细胞术(FCM)检测B细胞表面共刺激分子的表达;CBA法检测培养上清中IL-6、IL-10、IL-12p70、IFN-γ和TNF浓度;利用相关的信号转导抑制剂检测参与B细胞内细胞因子分泌的信号转导通路,探讨TLR9激动剂CpG ODN刺激小鼠B细胞后,对B细胞表面共刺激分子表达和细胞因子分泌的影响。结果表明:CpG ODN可以上调B细胞表面CD40、CD80、CD86和MHC II类分子的表达,促进IL-6、IL-10和TNF的高分泌。JNK、p38和NF-κB信号转导通路调控B细胞IL-6、IL-10和TNF的分泌。说明CpG ODN可以通过上调B细胞共刺激分子表达和促进细胞因子分泌等多方面调节B细胞功能。Freshly purified splenic B cells were stimulated with or without CpG ODN. After 24 h, cells were analyzed for costimulatory molecules expression by FCM and culture supernatants were collected for the measurement of IL-6, IL-10, IL-12p70, IFN-y and TNF by CBA. PD98059 (ERK inhibitor), SP600125 (JNK inhibitor), SB203580 (p38 inhibitor) and PDTC (NF-tcB inhibitor) were used to study the signal pathways for the induced IL-6, IL-10 and TNF secretion. To study the effects of CpG ODN (TLR9 agonist) onthe expression of co-stimulatory molecules and cytokines production in B cells. Results showed that CpG ODN promoted up-regulated the expression of CD40, CD80, CD86 and MHC class II, and promoted preferential IL-6, IL-10 and TNF secretion by B cells. Furthermore, activation of JNK, p38 and NF-tcB was essential for IL-6, IL-10 and TNF production by B cells after CpG ODN stimulation. These results suggest that TLR9 agonist has pleiotropic effects on B cells involving cytokine secretion and expression of costimulatory molecules.
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