人淋巴细胞H9体外感染HIV-1后APOBEC3G mRNA水平的监测

目的：观察人淋巴细胞H9体外感染HIV-1后,在其细胞内APOBEC3G蛋白水平降低的同时,APOBEC3GmRNA的水平是否也下降,阐明HIV-1抵抗APOBEC3G蛋白抗病毒作用的机制。方法：H9细胞感染HIV-1后,在不同时间点收集细胞和培养上清,提取细胞总RNA,应用实时定量PCR检测APOBEC3G mRNA的水平。检测上清液的HIV-1 p24抗原含量验证H9细胞是否感染了HIV-1。结果：（1）采用？？CT相对定量PCR方法检测APOBEC3G mRNA水平,其准确性高于94%。（2）在体外水平,H9细胞在感染HIV后,APOBEC3G mRNA水平没有显著变化（P＆gt;0.05）。结论：HIV-1可能主要通过降低APOBEC3G蛋白的水平而抵抗其抗病毒作用,这比通过降低mRNA水平来抵抗APOBEC3G的抗病毒作用更为快速、有效。

Monitoring of the APOBEC3G mRNA level of human lymphocyte H9 with HIV-1 infection in vitro

Objective：To find out whether APOBEC3G mRNA levels of H9 cells decline with the decrease in protein levels after human lymphocyte H9 was infected with HIV-1 in vitro, and to elucidate the mechanism by which HIV-1 count- er acts the antiviral effect of APOBEC3G. Methods：After H9 cells were infected with HIV-l,cells and culture supernatant were collected at different time points. Total RNA was extracted. Real-time PCR was used to test APOBEC3G mRNA lev- els, In order to determine whether 1-19 cells were infected by HIV-1, the concentration of HIV-1 p24 antigen in the superna- rant was detected. Results： （1） The accuracy of measuring APOBEC3G mRNA level using △△ CT relative quantification method was over 94%. （2）The APOBEC3G mRNA level of H9 cell did not change after HIV-1 infection in vitro（P 〉 0.05）. Conclusion：It is possible that HIV-1 counteracts the antiviral effect of APOBEC3G mainly by down-regulating the APOBEC3G protein level. This is more quick and effective than down-regulating the APOBEC3G mRNA level.