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目的探讨β2糖蛋白I(β2GPI)免疫小鼠引起的辅助性T(Th)细胞分化和抗β2GPI抗体产生的机制。方法将健康的BALB/c小鼠随机分为生理盐水组和β2GPI免疫组,每隔2周由尾静脉注射β2GPI或等量生理盐水,共注射2次或4次。采用间接ELISA检测小鼠血清中抗β2GPI抗体的效价;实时荧光定量PCR(qRT-PCR)检测小鼠脾脏细胞GATA结合蛋白3(GATA3)、白细胞介素4(IL-4)、表达于T细胞的T盒(T-bet)、γ干扰素(IFN-γ)、Foxp3的mRNA水平;流式细胞术检测小鼠脾脏淋巴细胞中GATA3阳性细胞和Foxp3阳性细胞的比例。结果 2次免疫后,小鼠血清中出现高效价(1∶100 000)的抗β2GPI抗体,脾脏中Th2特异性指标GATA3、IL-4的mRNA水平和GATA3阳性细胞比例相对生理盐水组升高。4次免疫后,小鼠血清中抗β2GPI抗体效价进一步升高(>1∶100 000),脾脏GATA3、Th1细胞特异性标志物(T-bet、IFN-γ)和调节性T细胞特异性标志Foxp3的mRNA水平下调,IL-4的mRNA水平和2次免疫后无明显差异。此外,脾脏细胞中GATA3阳性细胞的比例进一步上调且伴有Foxp3阳性细胞比例的下调。结论β2GPI免疫小鼠引起Th细胞向Th2细胞优势分化,并伴有Th1细胞和调节性T细胞的抑制,有利于抗β2GPI抗体的产生。
Objective To investigate the mechanism of T (Th) cell differentiation induced by β2 glycoprotein I (β2GPI) immunized mice and the anti-β2GPI antibody production. Methods Healthy BALB / c mice were randomly divided into normal saline group and β2GPI immunization group, and were injected with β2GPI or normal saline intravenously every two weeks for two or four times. The titer of anti-β2GPI antibody in serum was detected by indirect ELISA. GATA3 and IL-4 were detected by real-time quantitative PCR (qRT-PCR) T-bet, IFN-γand Foxp3 mRNA levels were detected by flow cytometry. The proportion of GATA3-positive cells and Foxp3-positive cells in splenic lymphocytes was detected by flow cytometry. Results After two immunizations, anti-β2GPI antibody with high titer (1: 100000) appeared in the serum of mice. The level of GATA3, IL-4 mRNA and GATA3-positive cells in the spleen were increased compared with those in the normal saline group. After 4 immunizations, anti-β2GPI antibody titers were further increased (> 1: 100,000) in mouse serum, spleen GATA3, Th1 cell specific marker (T-bet, IFN- γ) and regulatory T cell specificity The mRNA level of Foxp3 was down-regulated, and the mRNA level of IL-4 and no significant difference after 2 immunizations. In addition, the proportion of GATA3-positive cells in spleen cells is further up-regulated and accompanied by the down-regulation of Foxp3-positive cell ratio. Conclusion The β2GPI immunized mice induced the predominance differentiation of Th cells to Th2 cells with the suppression of Th1 cells and regulatory T cells, which is favorable for the production of anti-β2GPI antibodies.