HIV-1感染对人CD4^+T淋巴母细胞内ITAMs/ITIMs通路的影响

Effects Of HIV-1 infection on related genes and proteins of ITAMs/ITIMs signaling pathway in human CD4^+T lymphoblast cells

目的探讨感染人免疫缺陷病毒1(HIV-1)后人CD4^+T淋巴母细胞(M8166细胞)内免疫受体酪氨酸激活基序(ITAMs)/免疫受体酪氨酸抑制基序(ITIMs)通路的变化。方法体外培养M8166细胞,随机分为空白组及感染组。空白组常规培养,感染组则以10 TCID50 HIV-1进行感染。用RT-PCR技术检测ITAMs/ITIMs信号通路相关受体基因的表达,以Western blotting法检测ITAMs/ITIMs信号通路相关受体蛋白及磷酸化蛋白的表达。结果与空白组比较,病毒组SHP-1 mRNA相对表达量高(P<0.01),SHIP1 mRNA低,但差异无统计学意义(P>0.05);两组SHP-2、SHIP2、Zap70、Syk mRNA相对表达量比较差异无统计学意义(P均>0.05)。与空白组比较,感染组SHP-1、p-SHP-1蛋白相对表达量高(P均<0.05),SHIP1、p-SHIP1蛋白相对表达量低(P均<0.05),Syk蛋白相对表达量高、p-Syk蛋白相对表达量低,但差异均无统计学意义(P均>0.05);两组SHP-2、SHIP2、p-SHIP2蛋白相对表达量比较差异无统计学意义(P均>0.05);两组均未见p-SHP2、Zap70、p-Zap70蛋白表达。结论 M8166细胞感染HIV-1后,其ITAMs/ITIMs信号通路被部分激活。

Objective To investigate the expression of related genes and proteins of immunoreceptortyrosine-based activation motifs（ ITAMs）/immunoreceptortyrosine-based inhibition motifs（ ITIMs） signaling pathway in human CD4^＋T lymphoblast cells（ M8166 cells） after HIV-1 infection. Methods M8166 cells cultured in vitro were randomly divided into the blank group and infection group. The cells in the blank group were normally cultured without special treatment,while the cells in the infection group were infected with 10 TCID50 HIV-1. The mRNA expression levels of related genes of ITAMs/ITIMs signaling pathway were detected by real-time fluorescent quantitative PCR,and the expression of the protein of related receptors was detected by Western blotting. Results Compared with the blank group,the expression of SHP-1 mRNA in the M8166 cells increased after HIV-1 infection（ P〈0. 01）,and the expression of SHIP-1 mRNA showed a decreasing trend but there was no statistical significance（ P〉0. 05）. No significant difference was found in the mRNA expression of SHP-2,SHIP2,Zap70,and Syk（ all P〉0. 05）. On the other hand,the expression of SHP-1 and P-SHP-1 protein significantly increased after HIV-1 infection（ both P〈0. 05）,and the expression of SHIP1 and P-SHIP1 proteins significantly decreased（ both P〈0. 05）. The expression of Syk was slightly up-regulated and P-Syk was down-regulated,but there was no statistical difference（ both P〈0. 05）. No significant difference was found in the expression of SHP-2,SHIP2 and P-SHIP2 between these two groups（ all P〉0. 05）,and no expression was found in P-SHP2,Zap70,and PZap70. Conclusion The ITAMs/ITIMs signaling pathway of M8166 cells is activated after HIV-1 infection.