HIV-INFECTED PROMYELOCYTIC CELL LINE

HIV感染的早幼粒细胞系

• 批准号: 3753443
• 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/3753443
• 关键词: HIV infections; cellular pathology; human immunodeficiency virus 1; immunofluorescence technique; leukopoiesis; neutrophil; tissue /cell culture; virus replication

The principal target cells of in vivo HIV infection are the CD4-positive T-cells and cells of the monocyte lineage. However, little is known about the role of other phagocytic cells (i.e. neutrophils) in AIDS. The neutrophil is a primary cellular component of host defense against bacterial infection (e.g. periodontal disease) and neutrophil dysfunction does occur in AIDS. The goal of this study was to examine the biological properties of the neutrophil-precursor HL-60 cell line after infection with HIV-1. To accomplish this, a chronically infected HL-60 culture was established using the HIIV-IIIb isolate. Productive infection with HIV was measured using a HeLa-CD4-ltr-beta gal indicator (MAGI) cell assay system and p24 production. Analysis of the infected HL-60 culture revealed that only 25% of the cells were infectious. Immunofluorescence (IF) studies suggested the presence of HIV proteins in about 50% of cells in this culture. Limiting dilution allowed for further isolation of two HIV-infected HL-60 subclones (see Table). Subclone #1 displayed a high level of infection by IF, but did not produce infectious virus. Subclone #22, as confirmed by methods of detection, appeared to be nearly 100% infected and capable of production/transmission of infectious HIV particles. Electron microscopy was used to show the presence of intact HIV particles. In situ hybridization (ISH) studies, using HIV-specific digoxigenin-labeled RNA probes, demonstrated high levels of viral mRNA intracellularly in the "parent' as well as in the two subclones. Culture MAGI%* p24(ug/ml)* IF(%)* ISH(%)* parent HL-60 25 138 50 60 subclone #1 0 127 90 60 subclone#22 100 400 95 100 *percentage of total cells estimated to be position. In conclusion, subclones #1 and #22 may serve as useful in vitro analogues for the study of the altered phagocytic response in the HIV-infected host. The establishment of the parameters concerning the HIV infection of this promyeloctic cell line may provide new knowledge into the cellular mechanisms of HIV pathogenesis in the host. Key words: HIV infection, HIV pathogenesis, HL-60, neutrophils, phagocytes

体内HIV感染的主要靶细胞是CD4阳性细胞 T细胞和单核细胞系的细胞。然而，人们对此知之甚少 关于其他吞噬细胞(即中性粒细胞)在艾滋病中的作用。 中性粒细胞是宿主防御的主要细胞成分。 细菌感染(如牙周病)和中性粒细胞 艾滋病患者确实会出现功能障碍。这项研究的目标是检查 中性粒细胞前体细胞株HL-60的生物学特性 在感染HIV-1之后。为了做到这一点，一个慢性感染的人 用HIIV-IIIB分离株建立HL-60细胞培养体系。多产 使用HeLa-CD4-ltr-beta Gal指示剂测量HIV感染情况 (MAGI)细胞分析系统和p24的产生。对感染者的分析 HL-60细胞培养显示，只有25%的细胞是有感染性的。 免疫荧光(IF)研究表明存在HIV蛋白 在该培养物中约50%的细胞中。允许的限制稀释 进一步分离两个感染艾滋病毒的HL-60亚克隆(见表)。 亚克隆1通过IF表现出高水平的感染，但没有 产生传染性病毒。亚克隆#22，由以下方法确认 检测到，似乎几乎100%被感染，并能够 产生/传播具有传染性的艾滋病毒颗粒。电子 显微镜被用来显示完整的HIV颗粒的存在。在……里面 应用HIV特异性地高辛标记的原位杂交(ISH)研究 RNA探针显示，在细胞内有高水平的病毒mRNA “亲本”以及两个亚克隆中的。 培养MAGI%*p24(微克/毫升)*IF(%)*ISH(%)* 家长HL-60 25 138 50 60 亚克隆#1 0 127 90 60 亚克隆#22 100 400 95 100 *估计为仓位的单元格总数的百分比。在……里面 结论：亚克隆#1和亚克隆#22在体外可作为有用的类似物 用于研究HIV感染者的吞噬反应改变 主持人。HIV感染相关参数的建立 这一早幼粒细胞系的建立可能为进一步研究 HIV在宿主中致病的细胞机制。 关键词：HIV感染，HIV发病机制，HL-60，中性粒细胞， 吞噬细胞