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目的观察TLR3激动剂Poly I:C对B细胞功能的影响,包括细胞增殖、共刺激分子表达、细胞因子和抗体的分泌情况。方法利用免疫磁珠法分选小鼠脾脏CD19+B细胞,RT-PCR法证实其表达TLR3。体外用Poly I:C刺激B细胞一定时间后,CFSE分裂法检测B细胞增殖,流式细胞术(flow cytometry,FCM)检测B细胞表面共刺激分子表达,CBA(cytometric bead ar-ray)法或ELISA法检测培养上清中细胞因子含量,CBA法检测B细胞分泌Ig的亚型。结果 Poly I:C可以促进B细胞增殖,上调B细胞表面CD40、CD80、CD86和MHC II类分子的表达,促进IL-6和TNF-α的高分泌,诱导IgG1κ抗体的产生。结论 Poly I:C可以通过促进增殖、细胞因子分泌,诱导抗体产生和上调共刺激分子表达等多方面调节B细胞功能。
Objective To observe the effect of TLR3 agonist Poly I: C on B cell function, including cell proliferation, costimulatory molecule expression, cytokine and antibody secretion. Methods The spleen CD19 + B cells were sorted by immunomagnetic beads method. The expression of TLR3 was confirmed by RT-PCR. After B cells were stimulated with Poly I: C for a certain period of time, the proliferation of B cells was detected by CFSE, the expression of costimulatory molecules on the surface of B cells was detected by flow cytometry (FCM), the CBA (cytometric bead ar-ray) The cytokines contents in culture supernatants were detected by ELISA and the subtypes of Ig secreted by B cells were detected by CBA method. Results Poly I: C can promote the proliferation of B cells, upregulate the expressions of CD40, CD80, CD86 and MHC class II molecules on B cells, promote the secretion of IL-6 and TNF-α and induce the production of IgG1 κ antibodies. Conclusion Poly I: C can regulate B cell function by promoting proliferation, secretion of cytokines, inducing antibody production and up-regulation of costimulatory molecules.