Detection and enumeration of HIV-1-producing cells by ELISPOT (enzyme-linked immunospot) assay

J Acquir Immune Defic Syndr Hum Retrovirol. 1999 Apr 15;20(5):442-7. doi: 10.1097/00042560-199904150-00005.

Abstract

The enzyme-linked immunospot (ELISPOT) assay was adapted to detect and enumerate HIV-1-producing cells at the single cell level. With CEM cells or peripheral blood mononuclear cells (PBMC) infected in vitro with HIV-1, the ELISPOT assay detected cells that produced HIV-1 antigens and showed that between 5.4% and 9.5% of the p24 antigen-positive CEM cells and 11.1% to 23.6% of the p24 antigen-positive PBMC were productively infected. In HIV-1-infected patients in early stage of the disease and without antiretroviral therapy, up to 4.54 HIV-1-producing cells per 10(6) CD4+ T lymphocytes were detected in peripheral blood and up to 277.75 HIV-1-producing cells per 10(6) CD4+ T lymphocytes were detected in splenic lymphoid tissue. Our results indicate that the ELISPOT assay could represent a new tool to study HIV-1 replication in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • CD4-Positive T-Lymphocytes / virology*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • HIV Infections / blood*
  • HIV-1*
  • Humans
  • Reagent Kits, Diagnostic*
  • Tumor Cells, Cultured

Substances

  • Reagent Kits, Diagnostic