Objectives

Elite controllers (EC) are a rare group of individuals living with HIV-1 who naturally control HIV-1 replication to levels below the limit of detection without antiretroviral therapy (ART) and rarely progress to AIDS. The mechanisms contributing to this control remain incompletely elucidated. In the present study, we have assessed whether cellular host factors could modulate HIV-1 replication post-entry in a controller-discordant couple living with HIV-1.

Methods

CD4 T cells from a controller-discordant couple, one partner being an EC and the other an HIV-1 progressor (PR), and healthy controls (HC) were isolated, activated and infected with VSV-G pseudotyped yellow fluorescent protein-encoding single-round HIV-1 virus (HIV-YFP). Viral reverse transcripts, 2-LTR circles and integrated proviral HIV-1 DNA were monitored by quantitative PCR (qPCR) and integration sites were analysed. We further measured LEDGF/p75 and p21 mRNA expression levels by qPCR.

Results

Infection of activated CD4 T cells with HIV-YFP was reduced in EC compared with the PR partner, and HC. Evaluation of viral DNA forms suggested a block after entry and during the early steps of HIV-1 reverse transcription in EC. The integration site distribution pattern in EC, PR and HC was similar. The p21 expression in CD4 T cells of EC was elevated compared with the PR or HC, in line with previous work.

Conclusions

Our study suggests a reduced permissiveness to HIV-1 infection of CD4 T cells from EC due to a block of HIV-1 replication after entry and before integration that might contribute to the EC phenotype in our patient.

中文翻译：

---

控制者不一致夫妇中离体HIV-1感染的分析

目标

精英控制者（EC）是一群罕见的HIV-1感染者，他们自然将HIV-1复制控制在低于未使用抗逆转录病毒疗法（ART）的检测限之下的水平，并且很少发展为AIDS。仍未完全阐明造成这种控制的机制。在本研究中，我们评估了在携带HIV-1的控制者不一致的夫妇中，细胞宿主因素是否可以调节进入后的HIV-1复制。

方法

分离，激活并感染了VSV-G伪型黄色荧光蛋白编码单株，从一个控制器不一致的夫妇中分离出CD4 T细胞，其中一个伴侣是EC，另一个伴侣是HIV-1进展剂（PR），而健康对照（HC）全面的HIV-1病毒（HIV-YFP）。通过定量PCR（qPCR）监测病毒逆转录本，2-LTR环和整合的原病毒HIV-1 DNA，并分析整合位点。我们通过qPCR进一步测量了LEDGF / p75和p21 mRNA的表达水平。

结果

与PR伴侣和HC相比，EC减少了HIV-YFP对活化的CD4 T细胞的感染。病毒DNA形式的评估表明，进入后以及EC-1中HIV-1逆转录的早期过程中都存在阻断作用。EC，PR和HC中的整合站点分布模式相似。与先前的研究相一致，与PR或HC相比，EC的CD4 T细胞中的p21表达升高。

结论

我们的研究表明，由于进入后和整合前HIV-1复制受阻，可能会导致EC表型降低，因此EC对CD4 T细胞的HIV-1感染的允许性降低。