title: Application of a 5-bromo-2'-deoxyuridine ELISA for measuring the lymphoproliferative response to human cytomegalovirus in HIV-1-infected patients.
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.
authors.affiliation: Department of Microbiology, School of Medicine, University of Valencia, Av. Blasco Ibáñez 17, 46010, Valencia, Spain
authors.forename: Amparo
authors.initials: A
authors.lastname: Tamarit
authors.forename: Juan
authors.initials: J
authors.lastname: Alberola
authors.forename: María José
authors.initials: MJ
authors.lastname: Galindo
authors.forename: David
authors.initials: D
authors.lastname: Navarro
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000821:virology; D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology*; D001973:Bromodeoxyuridine; D003587:Cytomegalovirus / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015497:HIV-1; D006801:Humans; D007202:Indicators and Reagents; D008213:Lymphocyte Activation
chemical_list: D007202:Indicators and Reagents; D001973:Bromodeoxyuridine