The effect of fresh lymphocytes on increased sensitivity of hiv-1 isolation: A multicenter study

Homayoon Farzadegan, David Imagawa, Phalguni Gupta, Moon H. Lee, Lisa Jacobson, Alfred Saah, Kathie Grovit, Charles R. Rinaldo, B. Frank Polk

Research output: Contribution to journalArticlepeer-review

Abstract

A multicenter study was undertaken to determine the sensitivity and reproducibility of markers for human immunodeficiency virus type 1 (HIV-1) viral growth and the effect of various preparations of lymphocytes on the sensitivity of standard and routinely used procedures for HIV-1 isolation. In phase 1, cocultivated culture supernatants obtained from 10 HIV-1 cultures were transported to three Multicenter AIDS Cohort Study (MACS) Virology Laboratories. Three commercial HIV-p24 antigen capture (AC) tests and two reverse transcriptase (RT) assays were used to ascertain the replication of HIV-1. The Du Pont and Abbott AC assays were found to be most sensitive (85-100%), and the RT assay with 24-h incubation period had comparable sensitivity (75-100%). In phase II, the sensitivity of standard cocultivation procedure for HIV-1 isolation was compared using freshly phytohemaggluti-nin-P (PHA-P)-stimulated, stimulated-frozen, and frozen-thawed and then stimulated normal human peripheral blood mononuclear cells (PBMCs) as co-cultivating cells. Blood samples from 13 HIV-1 infected individuals with various CD4+ cell counts were cocultivated in each of the three MACS laboratories using one of the aforementioned normal PBMCs. The PHA-P-stimulated fresh normal PBMC showed a maximum isolation rate of 100% (13 of 13) with an average of 8 days to positivity. This rate of isolation was significantly greater than other rates using any one of the other PBMC preparations. These findings demonstrated that the use of freshly PH A-P stimulated PBMCs maximized HIV-1 isolation from blood when a sensitive HIV-1 p24 AC assay or RT assay with overnight incubation is employed for the detection of HIV in culture supernatant.

Original languageEnglish (US)
Pages (from-to)981-986
Number of pages6
JournalJournal of acquired immune deficiency syndromes
Volume3
Issue number10
StatePublished - Oct 1990

Keywords

  • HIV-1
  • Isolation procedures
  • Lymphocytes

ASJC Scopus subject areas

  • Infectious Diseases
  • Pharmacology (medical)

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