IngentaConnect Abnormal intracellular IL-2 and interferon-gamma (IFN-) productio...

Authors: Westby¹; Marriott¹; Guckian¹; Cookson¹; Hay²; Dalgleish¹

Source: Clinical & Experimental Immunology, Volume 111, Number 2, February 1998 , pp. 257-263(7)

Abstract:

We used three-colour cytometry to analyse intracellular cytokine production in activated whole blood cultures derived from patients with HIV-1 infection. We assessed mitogen-induced IL-2, IL-4 and IFN-γ production from T cells as possible markers of immune dysfunction. The percentages of T cells staining for IL-2 were significantly reduced in stimulated cultures from HIV⁺ individuals relative to normal controls (P < 0.0001); this reduction was observed in both the CD4⁺ and the CD8⁺ subsets. IL-2 production was significantly reduced in CD4⁺ T cells from HIV⁺ individuals clinically classified as symptomatics compared with HIV⁺ asymptomatics (P < 0.001); in addition, production of IL-2 inversely correlated with viral load (r² = 0.832). On the other hand, HIV⁺ individuals showed significantly more T cells staining positive for IFN-γ (P < 0.0001); subset analysis identified these T cells as CD8⁺. Increased IFN-γ production in the CD8⁺ T cell subset of HIV⁺ individuals correlated neither with clinical status nor with plasma viral load. IL-4 staining in activated T cells was low (< 5%) and no differences were observed between HIV⁺ and control groups. Three-colour FACS analysis of whole blood provides a sensitive, rapid and relatively easy means to detect cytokine profiles within T cell subpopulations. Only small volumes of blood are required (0.5 ml), since there is no need for cell isolation, making it more practical than ELISA or reverse transcriptase-polymerase chain reaction (RT-PCR) for the analysis of immune function in HIV⁺ individuals. This technique could therefore play a role in mapping the dynamics and extent of immune recovery in AIDS patients undergoing triple combination therapy.

Keywords: HIV; cell-mediated immunity; cytokines; flow cytometry; whole blood cultures

Document Type: Original article

DOI: 10.1046/j.1365-2249.1998.00505.x

Affiliations: 1: Division of Oncology, St George's Hospital Medical School, 2: Department of Genito-Urinary Medicine, St George's Hospital, London, UK

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Abnormal intracellular IL-2 and interferon-gamma (IFN-γ) production as HIV-1-associated markers of immune dysfunction