Isolation of human immunodeficiency virus Type 1 from peripheral blood monocytes

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Human immunodeficiency virus type 1 (HIV-1) can infect circulating peripheral blood monocytes and resting CD4+ T lymphocytes despite sustained suppression of plasma viremia to undetectable levels. These persistently infected cell populations pose a barrier for virus eradication by highly active antiretroviral therapy (HAART), and are a significant reservoir of HIV-1 contributing to viral rebound following cessation or failure of HAART. This chapter provides a protocol for isolating replication-competent HIV-1 from peripheral blood monocytes of HIV-1-infected individuals, including those with sustained plasma HIV-1 RNA levels below 50 copies/mL, by co-culture with CD8-depleted, phytohemagglutinin-activated donor peripheral blood mononuclear cells. In our laboratory, this protocol has the sensitivity to achieve a success rate of positive HIV-1 isolation in approx 70 of cases. The study of HIV-1 strains harbored by peripheral blood monocytes of patients undergoing HAART will contribute to the understanding of viral persistence in cellular reservoirs that impede effective HAART.
Original languageEnglish
Title of host publicationMethods in Molecular Biology - Human Retrovirus Protocols: Virology and Molecular Biology
EditorsTuofu Zhu
Place of PublicationUSA
PublisherHumana Press
Pages25 - 33
Number of pages9
ISBN (Print)9781588294951
Publication statusPublished - 2005

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