DEVELOPMENT OF A SENSITIVE QUANTITATIVE FOCAL ASSAY FOR HUMAN IMMUNODEFICIENCY VIRUS INFECTIVITY
DEVELOPMENT OF A SENSITIVE QUANTITATIVE FOCAL ASSAY FOR HUMAN IMMUNODEFICIENCY VIRUS INFECTIVITY
复制标题
DOI：
10.1128/jvi.62.10.3779-3788.1988
复制
发表时间：
1988-10-01
影响因子：
5.4
通讯作者：
WEHRLY, K
中科院分区：
文献类型：
--
作者：
CHESEBRO, B;WEHRLY, K
传染性人类免疫缺陷病毒（HIV）的准确和灵敏的定量一直难以实现。在这份报告中，一个定量局灶免疫测定（FIA）的HIV开发使用人HeLa细胞，使易受HIV感染的CD 4基因通过逆转录病毒载体的引入。通过使用人抗HIV抗体或对HIV特异性的小鼠单克隆抗体以及对小鼠或人免疫球蛋白特异性的第二荧光素或过氧化物酶缀合抗体来鉴定感染的细胞。该试验鉴定了感染野生型或培养适应性HIV分离株的细胞，并能够在106个细胞中检出1个阳性细胞。FIA在检测无细胞HIV方面也是有效的，与使用A3.01、CEM和其他人类白血病细胞的检测相反，FIA检测到大多数野生型HIV分离株。HIV中和作用可以通过使用FIA来确定，并且发现与HIV gp 120反应的两种单克隆抗体仅中和HIV的LAV-IIIB株。这些单克隆抗体以及获得性免疫缺陷综合征患者血清样本中的抗体能够抑制人类淋巴细胞悬浮培养物中HIV感染的传播，但不能抑制粘附在塑料培养皿上的CD 4阳性HeLa细胞中HIV感染的传播。
Accurate and sensitive quantitation of infectious human immunodeficiency virus (HIV) has been difficult to achieve. In this report, a quantitative focal immunoassay (FIA) for HIV was developed using human HeLa cells rendered susceptible to HIV infection by introduction of the CD4 gene via a retrovirus vector. Infected cells were identified by using human anti-HIV antibodies or mouse monoclonal antibodies specific for HIV together with secondary fluorescein- or peroxidase-conjugated antibody specific for mouse or human immunoglobulins. The assay identified cells infected with either wild-type or culture-adapted HIV isolates and was capable of detecting 1 positive cell in 106 cells. The FIA was also effective at detecting cell-free HIV, and in contrast to assays using A3.01, CEM, and other human leukemia cells, the FIA detected most wild-type HIV isolates. HIV neutralization could be determined by using the FIA, and two monoclonal antibodies reactive with HIV gp120 were found to neutralize only the LAV-IIIB strain of HIV. These monoclonal antibodies, as well as antibodies in serum samples from patients with acquired immune deficiency syndrome, were able to inhibit the spread of HIV infection in human lymphocyte suspension cultures but not in CD4-positive HeLa cells growing attached to plastic dishes.