Abstract
HIV can be recovered from infected patients at all stages of the disease spectrum. Typically, the quantity of biologically active virus, or viral protein, in body tissues is below the level of direct detection by either antigen capture or reverse transcriptase assays. Consequently, the virus must be expanded in culture. This may be achieved by the cocultivation of patient material with mitogen-stimulated peripheral blood mononuclear cells (PBMCs) from normal, healthy donors. These cocultures are then maintained by regularly scheduled interleukin-2 (IL-2) supplemented medium replacement, and the periodic addition of freshly stimulated normal donor PBMCs. During this cocultivation period, culture fluids are harvested at regular intervals and tested for the presence and subsequent replication of HIV. Cultures failing to demonstrate evidence of virus expression within 35 d are usually terminated.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Burke, D. S, Fowler, A.K., Redfield, R.R., Dilworth, S., and Oster, C.N. (1990) Isolation of HIV-1 from the blood of seropositive adults: patient stage of illness and sample inoculum size are major determinants of a positive culture. J. AIDS 3(12), 1159ā1167.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 1999 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Lane, J.R. (1999). Isolation and Expansion of HIV from Cells and Body Fluids by Coculture. In: Michael, N.L., Kim, J.H. (eds) HIV Protocols. Methods in Molecular Medicineā¢, vol 17. Humana Press. https://doi.org/10.1385/0-89603-369-4:3
Download citation
DOI: https://doi.org/10.1385/0-89603-369-4:3
Publisher Name: Humana Press
Print ISBN: 978-0-89603-369-6
Online ISBN: 978-1-59259-601-0
eBook Packages: Springer Protocols