Analysis of Multiply Spliced Transcripts in Lymphoid Tissue Reservoirs of Rhesus Macaques Infected with RT-SHIV during HAART
Analysis of Multiply Spliced Transcripts in Lymphoid Tissue Reservoirs of Rhesus Macaques Infected with RT-SHIV during HAART
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DOI：
10.1371/journal.pone.0087914
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发表时间：
2014-02-05
期刊：
影响因子：
3.7
通讯作者：
North, Thomas W.
中科院分区：
文献类型：
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作者：
Deere, Jesse D.;Kauffman, Robert C.;North, Thomas W.
高效抗逆转录病毒疗法（HAART）可以将感染者体内的人类免疫缺陷病毒1型（HIV-1）水平降低到无法检测的水平，但病毒并没有被根除。HAART期间病毒持续存在的机制尚不清楚，但已经确定了一些储存库，如潜伏感染的静息记忆CD 4（+）T细胞。在潜伏期期间，除了在病毒转录的起始和延伸步骤处的阻断之外，在病毒RNA（vRNA）的输出中存在阻断，导致多剪接转录物在细胞核中的积累。由多重剪接转录物编码的两个基因是达特和Rev，它们在病毒复制周期的早期是必不可少的，并且可能指示给定细胞群体中的感染状态。在这里，多重剪接转录物的水平与HAART治疗的RT-SHIV感染恒河猴的组织样本中含gag的RNA水平进行了比较。在TaqMan PCR测定法的开发过程中鉴定了剪接位点序列变异。在胃肠道和淋巴组织中检测到多剪接转录本，但在胸腺中未检测到。多剪接转录物的水平低于gag RNA的水平，并且两者都与血浆病毒载量相关。在血浆病毒载量的猕猴胃肠道样本中，多重剪接与gag RNA的比例最大
Highly active antiretroviral therapy (HAART) can reduce levels of human immunodeficiency virus type 1 (HIV-1) to undetectable levels in infected individuals, but the virus is not eradicated. The mechanisms of viral persistence during HAART are poorly defined, but some reservoirs have been identified, such as latently infected resting memory CD4(+) T cells. During latency, in addition to blocks at the initiation and elongation steps of viral transcription, there is a block in the export of viral RNA (vRNA), leading to the accumulation of multiply-spliced transcripts in the nucleus. Two of the genes encoded by the multiply-spliced transcripts are Tat and Rev, which are essential early in the viral replication cycle and might indicate the state of infection in a given population of cells. Here, the levels of multiply-spliced transcripts were compared to the levels of gag-containing RNA in tissue samples from RT-SHIV-infected rhesus macaques treated with HAART. Splice site sequence variation was identified during development of a TaqMan PCR assay. Multiply-spliced transcripts were detected in gastrointestinal and lymphatic tissues, but not the thymus. Levels of multiply-spliced transcripts were lower than levels of gag RNA, and both correlated with plasma virus loads. The ratio of multiply-spliced to gag RNA was greatest in the gastrointestinal samples from macaques with plasma virus loads