Fig. 5
From: Role of germinal center and CD39highCD73+ B cells in the age-related tonsillar involution
B-cells metabolically adapted to catabolize ATP increase with increasing age. (A) Fresh TMC were stained for surface CD20, CD3, CD73, CD39 and a dead/live stain. Samples were subsequently analyzed by FACS. The gating strategy to identify the B-cell population analyzed, is partially illustrated. Singlets were gated by plotting FSC-H vs. FSC-A for each sample (not shown). Within the singlets population, dead cells were determined by a viability dye (not shown). Within the viable gate, lymphoid gate was determined through SSC-A vs. FSC-A as well as the CD20 gate (not shown). Dot plots depicting the percentage of the tonsillar B cells co-expressing CD39high and CD73 (P1), CD20+CD39IntCD73− (P2) and CD20+CD39highCD73− (P3) populations of selected individuals, from the children (left) and teenagers group (right). (B) Histograms presenting the mean percentage ± SD of the CD20+CD39highCD73+ population frequencies determined as in A), from 69 individuals, distributed according their age. p value was calculated through unpaired t test, ***p < 0.005. Frequencies of the P2 and P3 cell populations were not recorded for this paper. (C) Fresh TMC were stained for surface CD20, CD4, CD8, CD73, CD39 and intra-nuclear Ki-67. Middle left hand panel: analysis was performed as in A), the P1, P2 and P3 cell populations from a donor are depicted. Right hand panels: representative dot plots of proliferating cell populations within the P1, P2 and P3 gates as indicated by the respective lines. Percentages designate frequencies of the populations indicated relative to their respective gate. Gates were manually adjusted due to the changes experienced by the cells in culture upon each treatment