6533b870fe1ef96bd12cfb54

RESEARCH PRODUCT

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

Amparo TamaritDavid NavarroJuan AlberolaMaría José Galindo

subject

Human cytomegalovirusCellular immunityvirusesCytomegalovirusEnzyme-Linked Immunosorbent AssayBiologyLymphocyte ActivationPeripheral blood mononuclear cellViruschemistry.chemical_compoundVirologymedicineHumansAcquired Immunodeficiency SyndromeAIDS-Related Opportunistic Infectionsvirus diseasesmedicine.diseaseVirologyDeoxyuridineBromodeoxyuridinechemistryCytomegalovirus InfectionsHIV-1Indicators and ReagentsThymidineLymphoproliferative responseBromodeoxyuridine

description

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.

yearjournalcountryeditionlanguage
2002-09-01Journal of Virological Methods
https://doi.org/10.1016/s0166-0934(02)00107-6