Isolation and propagation of HIV-1 on peripheral blood mononuclear cells
Isolation and propagation of HIV-1 on peripheral blood mononuclear cells
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DOI：
10.1038/nprot.2008.3
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Kootstra, Neeltje A.
中科院分区：
文献类型：
--
作者：
van 't Wout, Angelique B.;Schuitemaker, Hanneke;Kootstra, Neeltje A.
人类免疫缺陷病毒1型（HIV-1）感染的特征是CD 4 + T细胞和T细胞功能的逐渐丧失以及持续的高水平病毒复制。HIV-1逆转录酶的高复制率和易出错的性质在整个感染过程中产生了不同的病毒准种。为了研究HIV-1准种的生物学特性与临床感染过程的关系，体外保存病毒的表型特征是必不可少的。在这里，我们描述了散装分离的HIV-1准种的方法和有限稀释病毒分离协议，通过该协议，可以获得单个共存的HIV-1变体使用外周血单核细胞从健康供体作为靶细胞。此外，还提供了HIV-1的增殖和滴定方法。
Human immunodeficiency virus type 1 (HIV-1) infection is characterized by a gradual loss of CD4+ T cells and T-cell function and an ongoing high level of virus replication. The high replication rate and the error-prone nature of HIV-1 reverse transcriptase create a diverse viral quasispecies throughout infection. To study biological properties of HIV-1 quasispecies in relation to the clinical course of infection, the in vitro preservation of phenotypical characteristics of the virus is essential. Here, we describe the method for bulk isolation of the HIV-1 quasispecies and a limiting dilution virus isolation protocol by which single coexisting HIV-1 variants can be obtained using peripheral blood mononuclear cells from a healthy donor as target cells. In addition, methods for propagation and titration of HIV-1 are provided.