HIV-INFECTED PROMYELOCYTIC CELL LINE

HIV感染的早幼粒细胞系

• 批准号: 3775566
• 负责人: Michael C Lynch
• 金额: --
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3775566
• 关键词: HIV infections; cell cell interaction; cell population study; electron microscopy; enzyme linked immunosorbent assay; flow cytometry; genetic strain; helper T lymphocyte; human immunodeficiency virus; inflammation; mixed tissue /cell culture; myelocyte; virus antigen; virus replication

Characterization of an HIV-infected Promyelocytic Cell Line Previous studies have demonstrated the relative abilities of various HIV isolates to infect different cell lines. Here we describe the abilities of lymphocytropic HIV-IIIB and monocytropic HIV-MN strains to infect the promyelocytic HL-60 cell line and the maintenance of chronically infected cultures of low virus productivity with no apparent loss of viability. Using HIV- directed activation of galactosidase indicator cells (MAGI assay), monolayers of HeLa-CD4-LTR-~-gal cells revealed up to 15% of HL-60 cells were infected and this coculture produced large, multinuclear syncytia. Using p24 antigen ELISA, we observed significant core antigen production by day 12 post-infection for IIIB and by day 20 post-infection for MN. The HL-60/IIIB cultures, by flow cytometric analysis, revealed ability in anti-gp 120 binding, a decrease in anti-CD4 binding, an increase in FMLP receptor activity and that these infected cells were of smaller size and increased granularity. In higher titers of cell-associated virus samples, destruction of the cell monolayer was apparent, whereas little was noted in cell-free virus titers; this suggests a cell-cell virus transmission. Electron microscopy revealed a fusion-like event of cells in infected culture as well as the presence of large cytoplasmic vacuoles of unknown function. In the presence of dibutyryl cAMP, HIV-infected HL-60 cells were differentiated into neutrophil-like cells and this differentiation had no effect on infectivity of cultures. This may represent a useful in vitro model system for the study alterations in HIV-infected inflammatory cells. Future plans include the use of in situ hybridization techniques to detect early neff gene expression as a marker of infection for comparison to standard detection techniques, since studies have suggested tnis gene product, when detected in the absence of late viral proteins, may represent a restricted viral infection incapable of producing infective virus. Also, we plan to select for a clone to establish cultures of 100% infection in order to better study unique functional and morphologic changes characteristic of HIV-infection. Finally, development of a model system to examine cell-cell interactions between HIV-infected cells and endothelial cell cultures is planned. This may help to provide insight to the possibility of a role for HIV-infected cell-mediated tissue destruction in inflammatory processes. Key Words: HIV; HL-60, Inflammation

一株HIV感染的早幼粒细胞系的鉴定 之前的研究已经证明了 不同的HIV分离株感染不同的细胞系。 这里我们 描述了嗜淋巴细胞的HIV-IIIB和单核细胞的能力， HIV-MN株感染早幼粒细胞HL-60细胞系， 低病毒慢性感染培养物的维持 生产力，没有明显的活力损失。 使用艾滋病毒- 半乳糖苷酶指示细胞的定向活化（MAGI测定）， 单层HeLa-CD 4-LTR-~-gal细胞显示高达15%的HL-60 细胞被感染，这种共培养产生了大的，多核的 合体 使用p24抗原ELISA，我们观察到显著的核心 IIIB感染后第12天和第20天的抗原产生 感染后的MN。 流式细胞仪检测HL-60/IIIB细胞系 分析显示抗gp 120结合的能力， 抗CD 4结合、FMLP受体活性增加以及 这些受感染的细胞体积较小， 粒度 在较高滴度的细胞相关病毒样品中， 细胞单层的破坏是明显的， 在无细胞病毒滴度中注意到;这表明细胞病毒 传输 电子显微镜显示了一个融合样事件， 感染培养物中的细胞以及大的 功能不明的细胞质空泡。 存在下 二丁酰cAMP，HIV感染的HL-60细胞分化为 嗜中性粒细胞样细胞和这种分化没有影响 文化的感染力。 这可能代表了一种有用的体外 研究艾滋病毒感染者改变的模型系统 炎症细胞 未来的计划包括在原地使用 杂交技术检测早期neff基因表达， 用于与标准检测进行比较的感染标志物 技术，因为研究表明tnis基因产物，当 在不存在晚期病毒蛋白的情况下检测到，可能代表 不能产生传染性病毒的限制性病毒感染。 此外，我们计划选择一个克隆，以建立100%的文化， 感染，以便更好地研究独特的功能和 HIV感染的形态学变化特征。 最后， 开发一个模型系统来检查细胞-细胞相互作用 HIV感染细胞和内皮细胞培养物之间的关系， 计划好了 这可能有助于深入了解 HIV感染细胞介导的组织破坏在 炎症过程。 关键词：HIV; HL-60;炎症