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2型腺相关病毒在人淋巴细胞中的复制及其与HIV-1的相互作用。

Replication of adeno-associated virus type 2 in human lymphocytic cells and interaction with HIV-1.

作者信息

Mendelson E, Grossman Z, Mileguir F, Rechavi G, Carter B J

机构信息

Central Virology Laboratory, Chaim Sheba Medical Center, Tel-Hashomer, Israel.

出版信息

Virology. 1992 Apr;187(2):453-63. doi: 10.1016/0042-6822(92)90447-w.

Abstract

Adeno-associated virus (AAV) is a nonpathogenic parvovirus which normally requires helper adenovirus or herpes-virus for replication. We examined the growth of AAV type 2 in human lymphocytes and its possible interaction with HIV-1. Three B cell lines (CK-B, HS-2, and UC729) and four T cell lines (Molt-4, Jurkat, HUT78, and HUT78+HIV, which is persistently infected with HIV-1) were infected with AAV either in the presence or in the absence of adenovirus. AAV DNA was found in cells of all the lines following incubation with the virus, indicating absorption. AAV DNA replication occurred in most cell lines without particular preference for B or T cells, but only in the presence of helper virus, either adenovirus or Epstein-Barr virus. Expression of AAV proteins was examined by immunoblotting and ELISA, using sera specific for AAV Rep or capsid proteins. The level of AAV protein synthesis correlated with the efficiency of AAV DNA replication, and both varied between cell lines. The yield of infectious AAV was low in most cases, except in one T4 line (Jurkat), where AAV replication and protein synthesis in the presence of adenovirus were very extensive. In HUT78+HIV cells both adenovirus and AAV (in the presence of Ad2) replicated efficiently. The effects of adenovirus plus AAV coinfections on HIV-1 replication, measured by reverse-transcriptase (RT) activity, were mild. Infection with adenovirus or AAV alone resulted in a 60-70% increase in RT activity, while infection with AAV plus adenovirus resulted in a 20% decrease in RT activity. The yield of infectious AAV in this cell line was very low.

摘要

腺相关病毒(AAV)是一种非致病性细小病毒,通常需要辅助腺病毒或疱疹病毒才能进行复制。我们研究了2型腺相关病毒在人淋巴细胞中的生长情况及其与HIV-1的可能相互作用。在有或无腺病毒存在的情况下,用AAV感染了三种B细胞系(CK-B、HS-2和UC729)和四种T细胞系(Molt-4、Jurkat、HUT78以及持续感染HIV-1的HUT78+HIV)。在与病毒孵育后,在所有细胞系的细胞中都发现了AAV DNA,表明发生了吸附。AAV DNA复制发生在大多数细胞系中,对B细胞或T细胞没有特别偏好,但仅在有辅助病毒(腺病毒或爱泼斯坦-巴尔病毒)存在的情况下发生。使用针对AAV Rep或衣壳蛋白的血清,通过免疫印迹和ELISA检测AAV蛋白的表达。AAV蛋白合成水平与AAV DNA复制效率相关,且在不同细胞系之间有所不同。在大多数情况下,感染性AAV的产量较低,除了一个T4细胞系(Jurkat),在该细胞系中,腺病毒存在时AAV复制和蛋白合成非常广泛。在HUT78+HIV细胞中,腺病毒和AAV(在Ad2存在的情况下)都能高效复制。通过逆转录酶(RT)活性测量,腺病毒和AAV共感染对HIV-1复制的影响较小。单独感染腺病毒或AAV会导致RT活性增加60%-70%,而同时感染AAV和腺病毒会导致RT活性降低20%。该细胞系中感染性AAV的产量非常低。

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