Immune signatures and TCR clonality of the expanded Tregs before and after xenotransplantation into NSG-SGM3 mice. (A) CyTOF analysis shows the relative median expression of the 29 markers in HD (n = 3) and AA (n = 4) expanded total Tregs. (B) Heat map of similarity matrix between pre- (n = 4) and postinjection (n = 4) of HD-expanded Tregs based on 29 parameters, computed as Pearson correlation coefficient. (C) Heat map of similarity matrix between pre- (n = 3) and posttransplantation (n = 3) of AA-expanded Tregs into NSG-SGM3 mice based on 29 parameters, computed as Pearson correlation coefficient. (D) Heat map of CyTOF analysis shows the comparison of the median expression of markers that defined Treg-B (CD45RA, CD45RO, CD95, and CCR4) between pre- (n = 4) and posttransplanted (n = 4) HD-expanded Tregs. ***P ≤ .001. (E) Heat map shows the comparison of the median expression of Treg-B–specific markers (CD45RA, CD45RO, CD95, and CCR4) between pre- (n = 3) and posttransplanted (n = 3) AA-expanded Tregs. ***P ≤ .001. (F) Productive clonality of the TCR repertoires of AA-expanded Tregs before (n = 3) and after (n = 3) xenotransplantation into NSG-SGM3 mice. Values near 1 represent samples with 1 or a few predominant rearrangements (monoclonal or oligoclonal samples) dominating the observed repertoire. Clonality values near 0 represent more polyclonal samples. (G) Representative pie charts show the diversity of TCR Vβ CDR3 in AA-expanded Tregs before and after xenotransplantation into NSG-SGM3 mice.