Highly productive infection with pseudotyped human immunodeficiency virus type 1 (HIV-1) indicates no intracellular restrictions to HIV-1 replication in primary human astrocytes
Highly productive infection with pseudotyped human immunodeficiency virus type 1 (HIV-1) indicates no intracellular restrictions to HIV-1 replication in primary human astrocytes
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DOI：
10.1128/jvi.75.17.7925-7933.2001
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发表时间：
2001-09-01
影响因子：
5.4
通讯作者：
Volsky, DJ
中科院分区：
文献类型：
--
作者：
Canki, M;Thai, JNF;Volsky, DJ
人类星形胶质细胞在体内和体外都可以感染人类免疫缺陷病毒I型（HIV-1），但与T淋巴细胞和巨噬细胞相比，病毒的表达效率低下。为了研究人类胎儿星形细胞中HIV-1的生命周期，我们用两性性小鼠白血病病毒或水泡性口炎病毒的包膜糖蛋白感染HIV-1假型细胞。这两种假病毒的感染都是高效且持久的，在病毒接种后7天达到峰值，感染细胞68%，每ml培养上清中有1.7杯病毒p24，然后在7周的随访中病毒表达水平逐渐下降。相比之下，在星形胶质细胞感染天然HIV-1的高峰期，不到0.1%的HIV-1抗原阳性细胞和400 pg / ml的细胞外p24。感染细胞和对照细胞的细胞活力和生长动力学相似。Northern blot分析显示，在感染后2、14和28天，暴露于假型（而非野生型）HIV-1的星形胶质细胞中存在9、4和2 kb的主要HIV-1 RNA种类。与生产性感染一致，在4周的随访中，假型HIV-1感染的星形胶质细胞中9- kb和4-kb的病毒转录本与2-kb的mRNA一样丰富，并且通过免疫印迹或细胞染色在感染细胞中检测到结构和调节病毒蛋白。这些细胞释放的子代病毒具有传染性。这些结果表明，原代星形胶质细胞感染HIV-1的主要屏障是病毒进入，星形胶质细胞对HIV-1的有效复制没有内在的胞内限制。
Human astrocytes can be infected with human immunodeficiency virus type I (HIV-1) in vitro and in vivo, but, in contrast to T lymphocytes and macrophages, virus expression is inefficient. To investigate the HIV-1 life cycle in human fetal astrocytes, we infected cells with HIV-1 pseudotyped with envelope glycoproteins of either amphotropic murine leukemia virus or vesicular stomatitis virus. Infection by both pseudotypes was productive and long lasting and reached a peak of 68% infected cells and 1.7 mug of viral p24 per ml of culture supernatant 7 days after virus inoculation and then continued with gradually declining levels of virus expression through 7 weeks of follow-up. This contrasted with less than 0.1% HIV-1 antigen-positive cells and 400 pg of extracellular p24 per ml at the peak of astrocyte infection with native HIV-1. Cell viability and growth kinetics were similar in infected and control cells. Northern blot analysis revealed the presence of major HIV-1 RNA species of 9, 4, and 2 kb in astrocytes exposed to pseudotyped (but not wild-type) HIV-1 at 2, 14, and 28 days after infection. Consistent with productive infection, the 9- and 4-kb viral transcripts in astrocytes infected by pseudotyped HIV-1 were as abundant as the 2-kb mRNA during 4 weeks of follow-up, and both structural and regulatory viral proteins were detected in infected cells by immunoblotting or cell staining. The progeny virus released by these cells was infectious. These results indicate that the major barrier to HIV-1 infection of primary astrocytes is at virus entry and that astrocytes have no intrinsic intracellular restriction to efficient HIV-1 replication.