TLR9激动剂CpG ODN对B细胞分泌细胞因子的影响

Effects of TLR9 agonist CpG ODN on the production of cytokines in B cells

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.