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. 2016 Oct 20;11(10):e0164543. doi: 10.1371/journal.pone.0164543

Fig 1. Increase of ex vivo CD4+ and CD8+ T subsets from BL patients at T1R.

Fig 1

(A) Methodology for analysis of lymphocytic subsets isolated from newly obtained peripheral blood (time zero, T0) in FACSAria flow cytometer. Exclusion of dead cells (kit Live/Dead) and determination of lymphocyte region (FSC-A versus SSC-A; first dot plot at left). Exclusion of cell clumps by FSC-H X FSC-A (second dot plot at left). Determination of CD3+/CD4+ or CD3+/CD8+ T cells regions, by using specific monoclonal antibodies and isotype control (third dot plot) and subset analysis by marking with anti-CD45RA Alexa Fluor 488 versus anti-CCR7 PerCP antibodies (first dot plot at right). Ex vivo frequencies of CD4+ (B) and CD8+ T subsets (C). Significantly increased ex vivo frequency of double positive CD4+/CLA+ and CD8+/CLA+ T cells in T1R patients (D). Results represent a median ± SEM of isolated experiments from T1R (n = 12), BL (n = 10) and HV (n = 10). Significance levels are shown by the graphs, being **p<0.01 and ***p<0.001. Mann–Whitney test; comparison between groups.