Isolation of human immunodeficiency virus type 1 from peripheral blood monocytes.
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.
Gorry
Paul R
PR
Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Victoria, Australia.
Sonza
Secondo
S
Kedzierska
Katherine
K
Crowe
Suzanne M
SM
D016827:CD8 Antigens / Q000276:immunology; D018920:Coculture Techniques / Q000379:methods; D005434:Flow Cytometry; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000821:virology; D009000:Monocytes / Q000276:immunology / Q000821:virology*
D016827:CD8 Antigens