The median number of CD3+ events captured ex vivo was 867.5 (IQR 280 -1955) and was similar to those captured at 37 °C, 4 °C and at room temperature, but higher than those captured after thawing (p=0.007). selleck chemicals Statistical analyses were performed using GraphPad Prism 5 (San Diego, California, USA). Shapiro–Wilks test for normality was applied to determine the distribution of the grouped samples. Mann–Whitney U test was applied for nonparametric independent sample comparisons and Wilcoxon

signed rank tests were applied to matched samples for nonparametric comparison. Kruskal–Wallis ANOVA tests were used for non-parametric assessments of variation between groups, with Dunn’s post test applied to test for the effect of multiple comparisons. For comparison of frequencies, the X2 test was used to compare groups. All tests were two-tailed and p-values of < 0.05 were considered significant. Cervical cytobrush samples from 183 HIV-infected, therapy naïve women were included in this study to compare alternative conditions for transporting and storage of cervical cytobrushes from field clinic to laboratory to preserve cervical cell yields, viability and function. Table 1 describes the cohort and conditions evaluated. Of these 183 cervical cytobrushes, 113/183 were evaluated immediately (Group 1 ex vivo;

within 6 h of sampling at the clinic) while 70/183 were randomly assigned into four groups to investigate the effect of mock transport or storage on cell recovery and function. Groups 2–4 cytobrushes were incubated at BYL719 concentration 37 °C (27/183), 4 °C (5/183) or room

temperature (25/183) for 24 h prior to processing and analysis. Group 5 cytobrushes were processed and immediately frozen in liquid nitrogen (13/183). The median age of the women was 34 years (IQR 31–39) and there was no significant difference in the ages of the women in each of the five groups (p = 0.74). The median CD4 count of the HIV-infected women was 434 cells/mm3 (IQR 312–608.8) and the median log plasma viral load of the HIV-infected women was 3.7 (IQR 1.7–4.7). There was no significant difference in CD4 counts and plasma viral load between the groups. CD3 T cell yields from cervical cytobrush specimens processed immediately were compared with those processed after 24 h (Groups 2–4; Table 2). A median of 65 416 (IQR 23 424–14 4720) CD3+ T cells Avelestat (AZD9668) were obtained from cytobrushes processed ex vivo. Cervical CD3+ T cell counts obtained from cytobrushes processed after 24 h and maintained at 37 °C, 4 °C, or room temperature did not differ significantly from T cell counts measured ex vivo (p = 0.10), indicating that T cell numbers were relatively stable over 24 h. Furthermore, none of the cytobrushes evaluated in the delayed processing experiments became contaminated during the 24 h of study. Cervical cytobrush-derived CD3+ T cells retained a median of 99.5% (IQR 96.2–100.0%) viable cells at isolation (Table 2).