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. 2021 Jul 19;17(7):e1009732. doi: 10.1371/journal.ppat.1009732

Fig 7. Specific ADA-1 blockade attenuates healthy tonsillar GC-Tfh cell function.

Fig 7

We sorted GC-Tfh, pre-Tfh and autologous GC-B cells from the tonsils of healthy individuals and co-cultured GC-Tfh:GC-B as well as pre-Tfh:GC-B cells in a 1:1 ratio in the presence of SEB (SEB) with or without 10uM of the specific ADA-1 inhibitor EHNA (EHNA). (A-B) We collected culture supernatants 5 days after co-culture and analyzed the levels of total IgG in ng/ml by ELISA. (A) Total IgG level in supernatant from GC-Tfh:GC-B cell co-cultures. (B) Total IgG level in supernatant from pre-Tfh:GC-B cell co-cultures. Results are from 2 independent experiments (n = 6) and are represented as mean as well as before and after treatment lines. Data was analyzed with the two-tailed paired non-parametric Student’s t-test using the Wilcoxon matched-pairs signed rank test. (C-L) We collected culture supernatants 5 days after co-culture and analyzed GC-Tfh and pre-Tfh cell cytokine and chemokine levels by the Luminex assay. Levels of (C) BLC, (D) IL-21, (E) IL-10, (F) IL-4, (G) IFN-γ, (H) IL-17A, (I) IL-2, (J) IP-10, (K) MIP-1α and (L) MIP-1ß are shown in pg/ml. Results are from 2 independent experiments (n = 6) and are represented as mean ± SEM. Data was analyzed with the two-tailed unpaired non-parametric Student’s t-test using the Mann-Whitney test. Nominal p-values p<0.05 were considered statistically significant. * p<0.05, ** p<0.01 and *** p<0.001.