Generation of representative primary virus isolates from blood plasma after isolation of HIV-1 with CD44 MicroBeads
Generation of representative primary virus isolates from blood plasma after isolation of HIV-1 with CD44 MicroBeads
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DOI：
10.1007/s00705-010-0806-x
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发表时间：
2010-12-01
影响因子：
2.7
通讯作者：
van der Kuyl, Antoinette C.
中科院分区：
文献类型：
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作者：
Cornelissen, Marion;Heeregrave, Edwin J.;van der Kuyl, Antoinette C.
用从艾滋病毒感染者身上分离的无细胞病毒感染细胞培养物是出了名的困难，并导致病毒变异的丧失。在这里，我们描述了来自PBMC、U87.CD4.CCR5和U87.CD4.CXCR4细胞培养的病毒序列，并将它们与使用CD44微球从12名患者的血浆中分离出的病毒颗粒的病毒序列进行比较。在PBMC和U87、CD4、CCR5两种培养物中，培养后血浆病毒株回收率均为66%。此外，根据env-V3序列预测了辅助受体的使用，并在表达CCR5或CXCR4的U87细胞中进行了测试。使用CXCR4的病毒的回收率较低。只有50%的预测使用CXCR4的病毒簇可以从细胞培养中提取，而71%的使用CCR5的毒株在U87中发现。因此，用CD44微球分离原代病毒在细胞培养中能够很好地表现体内的差异。
Infection of cell cultures with cell-free virus isolated from HIV-infected patients is notoriously difficult and results in a loss of viral variation. Here, we describe viral sequences from PBMC, U87.CD4.CCR5 and U87.CD4.CXCR4 cell cultures and compare them to those from blood plasma from 12 patients from whom virus particles were isolated using CD44 MicroBeads. In both PBMC and U87.CD4.CCR5 cultures, 66% of the plasma viral strains were retrieved after culturing. In addition, coreceptor use was predicted based on the env-V3 sequence and tested in U87.CD4 cells expressing either CCR5 or CXCR4. Recovery was lower for the CXCR4-using viruses. Only 50% of the virus clusters predicted to use CXCR4 could be retrieved from cell cultures, while 71% of CCR5-using strains were found in U87.CCR5 cultures. Therefore, isolation of primary viruses with CD44 MicroBeads results in a good representation in cell culture of the in vivo divergence.