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The accuracy of these phenotypes has been recently reported [19]. Cellular activation in CD4 subsets was characterized by HLA-DR expression [20]. Senescent CD4+ T cells were characterized as CD28??CD57+. Proliferation levels were determined by intracellular immunostaining with Ki67. Cells were permeabilized using the fixation/permeabilization staining set (eBioscience, San Diego, CA, USA) according to the manufacturer��s instructions. Data on a minimum of 100?000 events were acquired. Acquisition was performed on a Cytomics FC500 flow cytometer and analysis used CXP software (Beckman Coulter). All continuous variables were expressed as a median for each variable (interquartile range), and categorical variables as the number of cases and percentages. Differences among categorical variables were analysed using the chi-squared test. The Mann�CWitney S6 Kinase non-parametric U test was used to analyse differences between continuous variables. Correlations between quantitative parameters were explored using the Spearman��s rho correlation coefficient. Statistical analysis was performed using Statistical Package for the Social Sciences software (SPSS 17, Chicago, IL, USA). A p value Lapatinib in vivo characteristic are shown in Table?1. All vertically HIV-infected subjects except one were under HAART at the time of study. HIV-infected and healthy subjects were similar in age and gender. A total of 25 (43.9%) vertically HIV-infected subjects had a detectable HIV viral load. Median CD4 T-cell counts (absolute and percentage) were significantly lower in vertically HIV-infected subjects who presented with a detectable viral load. No further LY294002 in vivo differences in clinical and demographic characteristics were observed between vertically HIV-infected subjects with detectable and undetectable viral loads. Naive and memory CD4+ T cells were significantly increased in vertically HIV-infected subjects when compared with healthy subjects (Table?2 and Fig.?1a,b). Interestingly, similar frequencies of effector memory and TemRA CD4 T cells were observed in both groups (Table?2). Taking advantage of the fact that in subjects who had been vertically infected the age matched the time of HIV infection, we analysed associations with age between vertically HIV-infected and healthy subjects. In this regard, significant negative correlations between naive CD4 T-cell frequency and age were observed in both healthy and vertically HIV-infected subjects (Fig.?2a,b). The opposite was found in memory and effector memory T-cell subsets. No correlation was observed between memory T-cell frequency and age in healthy subjects (Fig.