A sensitive microtiter infectivity assay for macrophage-tropic and primary isolates of HIV-1 based on the transactivation of a stably integrated gene for the green fluorescent protein.

Important note: Information in this article was accurate in 1997. The state of the art may have changed since the publication date.

A sensitive microtiter infectivity assay for macrophage-tropic and primary isolates of HIV-1 based on the transactivation of a stably integrated gene for the green fluorescent protein.

Conf Adv AIDS Vaccine Dev. 1997 May 4-7;:19. Unique Identifier : AIDSLINE MED/97927011
Tan GT; Honnen WJ; Kayman SC; Pinter A; Laboratory of Retroviral Biology, Public Health Research Institute,; New York, NY. Fax: (212) 578-0804.

Abstract: Reporter cell lines for the quantitative assay of HIV infectivity were developed using HOS cells rendered susceptible to HIV infection by transfection of genes for CD4, and either CCR5 or CXCR4 as the respective coreceptor for macrophage-tropic and T cell tropic HIV isolates. This 96-well microtiter assay is based on the transactivation of a stably integrated HIV-1 LTR-green fluorescent protein (GFP) transcription unit. Upon HIV-1 entry into these HOS target cells, Tat expression increased the HIV LTR-directed transcription of the GFP gene as demonstrated by the increased fluorescence of detergent lysates of infected cells relative to that of uninfected controls. The CCR5-containing cell line efficiently supported infection by a diverse array of macrophage-tropic viruses and clinical isolates. While a four-day assay was sufficiently sensitive in detecting infection mediated by concentrations of HIV-1 (Ba-L) as low as 1 ng/ml p24, detectable responses for 7 clinical isolates required 5 ng/ml p24. The assay was also adequately quantitative to reveal subtle differences in the infectivity levels of various virus isolates; primary isolates (10 ng/ml p24) typically induced a 2-3 fold fluorescence signal enhancement in four days, whereas an equivalent concentration of HIV-1(Ba-L), L consistently produced a 6-fold signal amplification. The assay's utility was demonstrated by the sensitive and accurate assessment of several HIV neutralizing monoclonal antibodies which gave predictable responses that paralleled those obtained in traditional PBMC systems. Thus, this assay provides a reproducible and standardized means for the quantitation of HIV-1, and the rapid and inexpensive evaluation of humoral immune responses against clinically relevant HIV-1 isolates.

Keywords: *HIV-1/PATHOGENICITY *Luminescent Proteins/GENETICS *Macrophages/VIROLOGY *Tropism
971130
M97B1205

AEGiS is a 501(c)3, not-for-profit, tax-exempt, educational corporation. AEGiS is made possible through unrestricted funding from Boehringer Ingelheim, Bridgestone/Firestone Charitable Trust, Bristol-Myers Squibb Company, Elton John AIDS Foundation, Gill Foundation, the National Library of Medicine, Quest Diagnostics, Roche and Trimeris, and donations from users like you. Always watch for outdated information. This article first appeared in 1997. This material is designed to support, not replace, the relationship that exists between you and your doctor.

AEGiS presents published material, reprinted with permission and neither endorses nor opposes any material. All information contained on this website, including information relating to health conditions, products, and treatments, is for informational purposes only. It is often presented in summary or aggregate form. It is not meant to be a substitute for the advice provided by your own physician or other medical professionals. Always discuss treatment options with a doctor who specializes in treating HIV.

Copyright ©1980, 1997. AEGiS. All materials appearing on AEGiS are protected by copyright as a collective work or compilation under U.S. copyright and other laws and are the property of AEGiS, or the party credited as the provider of the content. .