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. 2022 Aug 5;30(3-4):309–322. doi: 10.1038/s41434-022-00358-x

Fig. 5. Expanded HLA-A*02-specific CAR-Tregs show signs of senescence after extended in vitro culture.

Fig. 5

TX200-TR101 Tregs were cultured with high-dose IL-2 (1000 U/ml), either unstimulated or with TCR chronically stimulated with anti-CD3/CD28-coated dynabeads. 3 Treg donors were used with an average CAR transduction of 47 ± 3.0% (mean ± SEM) (A) Telomerase activity after 1, 22, and 43 days in culture, presented as mean % RTA ± SD. B Telomere length after 1, 22, and 43 days in culture, presented as mean telomere length (bp) ± SD. C Phenotypic analysis of sub-populations of CD4+ Tregs after 54 days of culture with IL-2 only or with anti-CD3/CD28-coated dynabeads plus IL-2 (means ± SEM). D Expression analysis of CD57 and/or KLRG1 cell surface markers on CD4+ Tregs after 54 days culture with IL-2 only or with anti-CD3/CD28-coated dynabeads plus IL-2 (means ± SEM). E Expression of SA-β-Gal activity in CD4+ Tregs after 54 days of culture with IL-2 only or with anti-CD3/CD28-coated dynabeads plus IL-2 (means ± SEM). Bars represent mean ± SD/SEM and each dot represents a Treg batch (n = 3 Treg batches, 5 technical replicates were assessed per batch).