Enhanced culture assay for detection and quantitation of latently infected, resting CD4+ T-cells carrying replication-competent virus in HIV-1-infected individuals

Methods Mol Biol. 2005;304:3-15. doi: 10.1385/1-59259-907-9:003.

Abstract

Highly active antiretroviral therapy can decrease plasma HIV-1 levels to below the limit of detection. However, HIV-1 persists in latently infected resting-memory CD4+ T-cells carrying an integrated copy of the viral genome. The pool of latently infected cells is extremely stable and represents a major barrier to HIV-1 eradication. Identification and characterization of this reservoir required the development of methods for purifying resting CD4+ T-cells from HIV-1-infected individuals, activating the cells to induce virus production, and detecting and quantitating cells capable of releasing infectious virus. The development of an enhanced viral culture assay to quantitate the number of latently infected cells carrying replication competent virus is described here.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • CD4 Lymphocyte Count / methods
  • CD4-Positive T-Lymphocytes / pathology
  • CD4-Positive T-Lymphocytes / virology*
  • Cell Culture Techniques / methods
  • Cell Separation / methods
  • HIV Infections / immunology
  • HIV Infections / virology*
  • HIV-1 / physiology*
  • Humans
  • Lymphocyte Activation
  • Viral Load
  • Virus Cultivation / methods
  • Virus Latency / physiology*
  • Virus Replication / physiology*