Abstract

Rapid culture and quantitation of human immunodeficiency virus type 1 from patient cells without the use of mitogen-stimulated donor cells. P K Kim , S He and J L HO Division of International Medicine, Cornell University Medical College, New York, New York 10021, USA. ABSTRACT We report the development of a rapid, sensitive virus culture method for direct quantitation of human immunodeficiency virus (HIV) in peripheral blood mononuclear cells (PBMCs). This new method involves culturing 10(7) PBMCs from HIV-seropositive persons in 10 ml of medium containing phorbol 12-myristate 13-acetate and interleukin-2. Both agents stimulate cell activation and hence viral replication. Cell-associated virus and free virus are quantitated by a commercially available HIV p24 antigen capture enzyme immunoassay. Detection of cell-associated p24 antigen by flow cytometry was less sensitive than by the enzyme immunoassay. In this preliminary study, HIV was detected in 20 of 23 HIV-seropositive patients and in none of the 11 HIV seronegative low-risk individuals. One HIV-seronegative person with Guillain-Barré syndrome following high-risk activity was found to be rapid-HIV-culture positive. The overall sensitivity and specificity were 87 and 100%, respectively. By comparing the quantity of virus produced in infected cells with the amount of virus produced in chronically infected U1 monocytes and ACH-2 lymphocytes stimulated with phorbol 12-myristate 13-acetate and interleukin-2, the approximate number of infected cells per sample is calculated. In the same patient specimens, quantitation of the number of HIV infected cells by the HIV rapid-culture method correlated with the results of the 21-day cell dilution coculture assay (correlation coefficient r = 0.5; 95% confidence interval, 0.07 to 0.77). Advantages of the rapid HIV culture include no requirement for donor PBMCs or change of media, shortened culture time, and the ability to detect p24 viral antigen from cell-associated virus for quantitation of viral load. CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? « Previous | Next Article » Table of Contents This Article Clin Vaccine Immunol November 1994 vol. 1 no. 6 660-666 » Abstract PDF Services Email this article to a colleague Similar articles in ASM journals Alert me when this article is cited Alert me if a correction is posted Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of CVI Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Load citing article information Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Kim, P. K. Articles by HO, J. L. Search for related content PubMed PubMed citation Articles by Kim, P. K. Articles by HO, J. L. Related Content Load related web page information Social Bookmarking CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? current issue November 2011, volume 18, issue 11 Alert me to new issues of CVI About CVI Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy CVI RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 1556-6811 Online ISSN: 1556-679X Copyright © 2011 by the American Society for Microbiology. For an alternate route to CVI .asm.org, visit: http://intl- CVI .asm.org | More Info» var gaJsHost = (("https:" == document.location.protocol) ? "https://ssl." : "http://www."); document.write(unescape("%3Cscript src='" + gaJsHost + "google-analytics.com/ga.js' type='text/javascript'%3E%3C/script%3E")); var pageTracker = _gat._getTracker("UA-5821458-5"); pageTracker._trackPageview();