Truckenmiller M E, Kulaga H, Coggiano M, Wyatt R, Snyder S H, Sweetnam P M
NovaScreen, Division of Nova Pharmaceutical Corporation, Baltimore, MD 21224.
AIDS Res Hum Retroviruses. 1993 May;9(5):445-53. doi: 10.1089/aid.1993.9.445.
HCN-1A is a human cerebral cortical neuronal cell line having properties consistent with cells of immature neuronal origin. This article details evidence for productive low-level infection of HCN-1A cells with human immunodeficiency virus type 1 (HIV-1). In vitro exposure to HCN-1A monolayers to a high titer of either LAV/HTLV-IIIB or HTLV-IIIMN resulted in HIV-1 p24 antigen production and a moderate increase in reverse transcriptase activity in cell-free supernatants. The cells in both LAV/HTLV-IIIB- and HTLV-IIIMN-infected cultures were passaged and proliferated as long as 5 weeks while continuing to express low levels of viral antigen. Virus-positive cells were detected by indirect immunofluorescence, using serum from an individual with acquired immune deficiency syndrome (AIDS) as well as with a gp120 monoclonal antibody. Confirmation of HCN-1A infection was provided by polymerase chain reaction analyses of both nuclear and cytoplasmic DNA and by de novo synthesis of viral proteins as shown by metabolic labeling and immunoprecipitation. Virus in cell-free supernatants from infected HCN-1A cultures was passaged to a permissive human T cell line (A3.01). HCN-1A cells had no detectable surface CD4 protein or CD4 message. However, the cells expressed the membrane glycolipids, galactocerebroside and sulfatide, possible receptors for gp120 on cells of neuronal origin. Undifferentiated HCN-1A cells provide an in vitro model for investigating potential interactions of HIV-1 with a homogeneous population of immature cortical neurons.
HCN - 1A是一种人类大脑皮质神经元细胞系,其特性与未成熟神经元来源的细胞一致。本文详细阐述了1型人类免疫缺陷病毒(HIV - 1)对HCN - 1A细胞进行有效低水平感染的证据。在体外,将高滴度的LAV/HTLV - IIIB或HTLV - IIIMN接种到HCN - 1A单层细胞上,导致HIV - 1 p24抗原产生,并且无细胞上清液中的逆转录酶活性适度增加。在LAV/HTLV - IIIB和HTLV - IIIMN感染的培养物中的细胞传代并增殖长达5周,同时持续表达低水平的病毒抗原。使用获得性免疫缺陷综合征(AIDS)患者的血清以及gp120单克隆抗体,通过间接免疫荧光检测到病毒阳性细胞。通过对核DNA和细胞质DNA的聚合酶链反应分析以及代谢标记和免疫沉淀所示的病毒蛋白的从头合成,证实了HCN - 1A感染。来自感染的HCN - 1A培养物的无细胞上清液中的病毒被接种到允许性人类T细胞系(A3.01)中。HCN - 1A细胞没有可检测到的表面CD4蛋白或CD4信息。然而,这些细胞表达了膜糖脂、半乳糖脑苷脂和硫脂,它们可能是神经元来源细胞上gp120的受体。未分化的HCN - 1A细胞为研究HIV - 1与同质的未成熟皮质神经元群体之间的潜在相互作用提供了一个体外模型。