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The role of T regulatory cells (Tregs) in human immunodeficiency virus (HIV)-1 infection, although not entirely clear, has recently been highlighted. Despite their lack of specificity, fluorochrome-labeled CD4 and CD25 antibodies are common flow cytometric reagents used to identify these cells with immunosuppressive potential. Cryopreservation has previously been shown to alter the proportions of lymphocytes with certain phenotypes expressed in peripheral blood mononuclear cells (PBMCs). The aim of this study was to assess the effect of cryopreservation on CD4+ CD25+ T cells in PBMCs from HIV-1+ individuals to guide the design of future studies on Tregs. We recruited 30 HIV-1+ individuals and nine healthy controls. CD25 expression in CD4+ T cells was compared between fresh and frozen/thawed PBMC samples from the same time point. In this study, cryopreservation significantly decreased the proportion of CD4+ CD25+ T cells in PBMC samples from HIV-1 infected subjects. This finding suggests that studies of CD4+ CD25+ T cells should be carried out on fresh samples to avoid bias introduced by cryopreservation.

Original publication

DOI

10.1002/jcla.20234

Type

Journal article

Journal

Journal of clinical laboratory analysis

Publication Date

01/2008

Volume

22

Pages

153 - 158

Addresses

Nuffield Department of Medicine, Oxford University, Oxford, United Kingdom.

Keywords

Leukocytes, Mononuclear, CD4-Positive T-Lymphocytes, Humans, HIV Infections, Cryopreservation, Flow Cytometry, Immunophenotyping, Cell Survival, Adolescent, Adult, Middle Aged, Child, Female, Male, T-Lymphocytes, Regulatory, Interleukin-2 Receptor alpha Subunit, Cold Temperature