人淋巴细胞H9体外感染HIV-1后APOBEC3G mRNA水平的监测  

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

作　　者：刘永健[1] 李林[1] 鲍作义[1] 刘思扬[1] 庄道民[1] 李韩平[1] 李敬云[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《军事医学科学院院刊》2009年第4期308-310,343,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金项目资助(30471496)

摘　　要：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>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。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.

关 键 词：HIV-1 APOBEC3G MRNA水平 

分 类 号：R512.91[医药卫生—内科学]