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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 observe whether APOBEC3G mRNA is downregulated in human cells after in vitro infection of human lymphocyte H9 with HIV-1, and to elucidate whether HIV-1 has an anti-viral mechanism against APOBEC3G protein. Methods: After H9 cells were infected with HIV-1, the cells and culture supernatants were collected at different time points to extract the total RNA of the cells. Real-time quantitative PCR was used to detect the level of APOBEC3G mRNA. Detection of HIV-1 p24 antigen content in supernatant Verify that H9 cells are infected with HIV-1. Results: (1) The accuracy of ?? APCTEC3G mRNA was detected by RT-PCR and the accuracy was higher than 94%. (2) There was no significant change in the level of APOBEC3G mRNA in H9 cells infected with HIV at the in vitro level (P> 0.05). CONCLUSION: HIV-1 may resist its antiviral effect mainly by decreasing the level of APOBEC3G protein, which is faster and more effective than the anti-virus effect of APOBEC3G by reducing mRNA level.