CD127 expression and regulation are altered in the memory CD8 T cells of HIV-infected patients--reversal by highly active anti-retroviral therapy (HAART) - PubMed (original) (raw)

Ex vivo expression of CD127 by memory CD8 lymphocyte taken from viraemic and highly active anti-retroviral therapy (HAART)-treated HIV-infected patients. Peripheral blood mononucear cells (PBMC) were incubated with antibody mixtures containing anti-CD8-APC (clone DK25, IgG1k, Dako Cytomation A/S, Glostrup, Denmark), anti-CD27-FITC (clone M-T27, IgG1k, Dako Cytomation S/A), anti-CD45RA Cy5 (clone HI 100, IgG2b, Pharmingen, San Diego, CA, USA) and anti-CD127-RPE (R34·34, IgG1, Immunotech, Marseille, France). The binding of anti-CD127 monoclonal antibodies (mAb) R34·34 to interleukin (IL)-7R alpha was not exposed to competition from IL-7 under our experimental conditions. After 30 min at 4°C, the cells were washed and fixed in phosphate-buffered saline (PBS) paraformaldehyde (1%). An isotype control was performed for each reagent. Staining analyses were performed on a fluorescence-activated cell sorter (FACSCalibur) flow cytometer running

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version 3·1 software (Becton Dickinson, Mountain View, CA, USA). Data were expressed as medians and interquartile ranges. Statistical analyses were performed by the Mann–Whitney _U_-test. The main comparisons were performed between healthy donors and viraemic patients, and between viraemic and HAART patients. (a) CD127 expression by CD8+ CD27+ lymphocytes. Naive cells (CD45RA+) and memory cells (CD45RA−). Results are shown for one representative individual in each group. CD45RA labelling delineated naive cells (CD45RA+) from memory CD8 lymphocytes (CD45RA−). (b) CD45RA and CD27 were used to delineate naive (CD45RA+ CD27+), memory (CD45RA− CD27+), effector (CD45RA− CD27−) and terminally differentiated effector lymphocytes (CD45RA+ CD27−). Proportion of CD127+ lymphocytes [mean fluorescence intensity (MFI) > 50] in the four CD8 lymphocyte subsets in healthy individuals, viraemic patients and HAART patients; n = 10 for each group. (c) Absolute counts per cubic millimetre for total CD127− lymphocytes (MFI < 50) in naive, memory, effector and terminally differentiated CD8 lymphocytes from each study group. □, Healthy donors; ▪ viraemic patients; , HAART patients. *P < 0·05 when compared to healthy donors; **P < 0·05 when compared to untreated viraemic patients.