In situ hybridization and immunocytochemistry for improved assessment of human immunodeficiency virus cultures.
In situ hybridization and immunocytochemistry for improved assessment of human immunodeficiency virus cultures.
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原位杂交和免疫细胞化学用于改进人类免疫缺陷病毒培养物的评估。
DOI：
10.1093/ajcp/88.6.673
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发表时间：
1987
影响因子：
3.5
通讯作者：
Vyas,GN
中科院分区：
文献类型：
--
作者：
Busch,MP;Rajagopalan,MS;Gantz,DM;Fu,S;Steimer,KS;Vyas,GN
作者描述了体外接种到敏感的人T细胞系和植物血凝素刺激的外周血单核细胞（PMC）培养物中后，与人类免疫缺陷病毒（HIV）复制有关的早期细胞内事件。感染后24小时内，用HIV DNA探针原位杂交检测到胞浆内的病毒RNA。在随后的2至10天内，通过采用单克隆抗体的免疫细胞化学测定法在感染细胞中检测到病毒核心抗原。几天后，通过逆转录酶测定和p25 gagantagen-capture测定检测无细胞病毒。当这些方法用于监测10个血清反应阳性者的PMC培养物时，病毒复制的类似进展是明显的：在感染后第3天，在PMC中检测到细胞质病毒RNA，随后出现细胞内病毒蛋白（第6-9天）和无细胞病毒（第12-21天）。原位杂交和免疫细胞化学提供了互补的，敏感的，和体外监测获得性免疫缺陷综合征病毒复制早期阶段的具体方法。
The authors characterized the early intracellular events involved in human immunodeficiency virus (HIV) replication afterin vitroinoculation into cultures of susceptible human T-cell lines and phytohemagglutinin-stimulated peripheralblood mononuclear cells (PMCs). Within 24 hours of infection, in situ hybridization with HIV DNA probe detected cytoplasmic viral RNA. Viral core antigen was detected in infected cells over the subsequent two to ten days by means of an immunocytochemical assay employing monoclonal antibodies. Several days later, cell-free virus was detected by both reverse transcriptase assay and a p25gagantigen-capture assay. When these methods were applied to monitor cultures of ten seropositive persons' PMCs, a similar progression of virus replication was apparent: cytoplasmic viral RNA was detected in infected PMCs by day 3, with the subsequent appearance of intracellular viral proteins (days 6-9) and cell-free virus (days 12-21).In situhybridization and immunocytochemistry offer complementary, sensitive, and specific approaches for monitoring the early stages of acquired immune deficiency syndrome virus replicationin vitro.