Application of a 5-bromo-2'-deoxyuridine ELISA for measuring the lymphoproliferative response to human cytomegalovirus in HIV-1-infected patients

J Virol Methods. 2002 Sep;105(2):247-51. doi: 10.1016/s0166-0934(02)00107-6.

Abstract

Assessment of the lymphoproliferative response to human cytomegalovirus (HCMV) may help to identify human immunodeficiency virus (HIV)-1-infected patients at high risk of developing HCMV end-organ disease. The tritiated thymidine ([3H]-TdR)-incorporation assay is the gold standard for measuring lymphoproliferative responses, though it is unsuitable as a routine laboratory procedure. An alternative non-radioactive technique, a 5-bromo-2'-deoxyuridine (BrdU) enzyme-linked immunosorbent assay, was applied for measuring T-cell proliferation in response to HCMV. Stimulation of either 1 x 10(5) or 5 x 10(4) peripheral blood mononuclear cells (PBMCs)/well with 10 PFU/well (before inactivation) of inactivated HCMV (AD169 strain) virions during 5 days, followed by an 18 h period of pulsing with BrdU (10 microM) proved to be the optimal laboratory conditions. The assay is simple, economical and feasible for monitoring the lymphoproliferative response to HCMV in HIV-1-infected patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS-Related Opportunistic Infections / virology
  • Acquired Immunodeficiency Syndrome / immunology*
  • Bromodeoxyuridine*
  • Cytomegalovirus / immunology
  • Cytomegalovirus / isolation & purification*
  • Cytomegalovirus Infections / diagnosis*
  • Cytomegalovirus Infections / immunology
  • Enzyme-Linked Immunosorbent Assay / methods*
  • HIV-1*
  • Humans
  • Indicators and Reagents
  • Lymphocyte Activation*

Substances

  • Indicators and Reagents
  • Bromodeoxyuridine