Detection of HIV-RNA-positive monocytes in peripheral blood of HIV- positive patients by simultaneous flow cytometric analysis of intracellular HIV RNA and cellular immunophenotype

Bruce K. Patterson*, Victoria L. Mosiman, Luis Cantarero, Manohar Furtado, Mondira Bhattacharya, Charles Goolsby

*Corresponding author for this work

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

Determinations of plasma HIV vital RNA copy numbers help to define the kinetics of HIV-1 infection in vivo and to monitor antiretroviral therapy. However, questions remain regarding the identity of various infected cell types contributing to this free virus pool and to the in vivo lifecycle of HIV during disease progression. Characterization of a novel fluorescence in situ hybridization (FISH) assay employing a pool of labeled oligonucleotide probes directed against HIV RNA was done followed by coupling of the FISH assay with simultaneous surface immunophenotyping to address these questions. In vitro characterizations of this assay using tumor necrosis factor-α stimulated and unstimulated ACH-2 cells demonstrated the ability to detect <5% HIV RNA positive cells with a sensitivity of <30 RNA copies per cell Peripheral blood mononuclear cells from 39 I-HIV-seropositive patients on no, single, combination, or triple drug therapy and 8 HIV-seronegative patients were examined. The majority of HIV-positive patients (24/39) harbored monocytes positive for HIV RNA and a significantly higher fraction of patients with high plasma vital load carried positive monocytes (13/16) than did patients in the low plasma viral load group (11/23). These results demonstrate the effectiveness of a novel FISH assay for identifying and monitoring HIV-infected cell populations in the peripheral blood of HIV- positive patients. In addition, monocytes are a major source of cellular HIV virus in the peripheral blood of HIV patients, even with progression of disease.

Original languageEnglish (US)
Pages (from-to)265-274
Number of pages10
JournalCytometry
Volume31
Issue number4
DOIs
StatePublished - Apr 1 1998

Keywords

  • HIV
  • Immunophenotyping
  • In situ PCR
  • In situ hybridization
  • Monocytes
  • RNA

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

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