Department of Microbiology and Medical Zoology, University of Puerto Rico, Medical Sciences Campus, San Juan, Puerto Rico.
J Leukoc Biol. 2010 Apr;87(4):633-6. doi: 10.1189/jlb.0809556. Epub 2009 Dec 30.
HIV-infected PM show restricted replication as compared with MDM. We aimed to determine at what point in the viral replication cycle this restriction occurs in PM as compared with MDM. We performed Alu-LTR PCR for proviral DNA to detect differences in HIV integration, real-time RT-PCR to measure env and gag mRNA levels, and Western blot analysis to detect differences in viral protein expression. PM and MDM were infected with HIV-1 BaL, and DNA was extracted after 24 h and at 6 days p.i. for real-time PCR studies. At 6 and 12 days p.i., cells were lysed for Western blot analyses. We found no difference in viral integration between PM and MDM but significantly lower levels of viral protein gp120 in PM than in MDM. Real-time RT-PCR analyses revealed 24-fold less env mRNA and tenfold less gag mRNA in PM. These results suggest that HIV-1 restriction in PM occurs at the level of transcription. This study is significant, as it advances our understanding of HIV-1 infection in PM and its contribution to decreased in utero vertical transmission.
与 MDM 相比,HIV 感染的 PM 显示出受限的复制。我们旨在确定与 MDM 相比,PM 中病毒复制周期的哪个阶段会发生这种限制。我们进行了 Alu-LTR PCR 以检测前病毒 DNA 中的 HIV 整合差异,进行了实时 RT-PCR 以测量 env 和 gag mRNA 水平,并进行了 Western blot 分析以检测病毒蛋白表达的差异。PM 和 MDM 感染了 HIV-1 BaL,并在感染后 24 小时和 6 天提取 DNA 进行实时 PCR 研究。在感染后 6 天和 12 天,裂解细胞进行 Western blot 分析。我们发现 PM 和 MDM 之间的病毒整合没有差异,但 PM 中的病毒蛋白 gp120 水平明显低于 MDM。实时 RT-PCR 分析显示,PM 中的 env mRNA 减少了 24 倍,gag mRNA 减少了 10 倍。这些结果表明,PM 中的 HIV-1 限制发生在转录水平。这项研究意义重大,因为它增进了我们对 PM 中 HIV-1 感染及其对降低宫内垂直传播的贡献的理解。