Fig. 6. expBreg can prolong allograft survival in a humanised mouse model of human skin transplantation.

a Kaplan–Meier survival curve demonstrating human allograft survival in a humanised mouse model of skin transplantation. Balb/c Rag2^−/− cy^−/− mice which were transplanted with human skin allograft are reconstituted with human PBMC alone or ±non-expanded CD19⁺ B cells or expBreg from a different human donor. n = 6 per group per experiment, one of three independent experiments is shown. Experimental schematic is presented in Supplementary Fig. 8a. b Summarised data of number of huCD45⁺CD4⁺ T cells and huCD45⁺CD20⁺ B cells of mice. n = 6 per group per experiment, one of three independent experiments is shown. Each dot is an individual mouse response. Gating strategy and representative histograms of live huCD45⁺ cells are presented in Supplementary Fig. 8b. c Summarised data of % expression of huCD45⁺CD20⁺TIM-1⁺ B cells in mice receiving PBMC + expBreg. n = 6 per group per experiment, one of three independent experiments is shown. Each dot is an individual mouse response. Representative FACS plots of live huCD45⁺CD20⁺TIM-1⁺ B cells are presented in Supplementary Fig. 8d. d Summarised data of serum levels of human IL-10, IL-2 and IL-17a in blood of mice. n = 6 per group per experiment, one of three independent experiments is shown. Each dot is an individual mouse response. e Summarised data of % live huCD45⁺CD4⁺CD25⁺CD127^lo Treg of mice. n = 6 per group per experiment, one of three independent experiments is shown. Each dot is an individual mouse response. Representative FACS plots of live huCD45⁺CD4⁺CD25⁺CD127^lo T cells are presented in Supplementary Fig. 8f. f Summarised data of % inhibition of CD4⁺ T-cell proliferation by human CD4⁺CD25⁺CD127^lo Treg which were generated in the presence or absence of expBreg. Data from experiments performed with cells from different healthy donors (n = 3) are presented. Each dot is an individual response. Representative FACS plots of live CD4⁺CD25⁺CD127^lo T cells are shown in Supplementary Fig. 8g. g Summarised data of % live CD4⁺CD25⁺CD127^lo human Treg on day-5 of suppression assay when autologous CD4⁺ T cells were co-cultured with anti-CD3/CD28 beads ± Isotype-expBreg, CD154-blocked-expBreg, IL-10-blocked-expBreg or TIM-blocked-expBreg. Data from experiments performed with cells from different healthy donors (n = 4) are presented. Each dot is an individual response. In (a–e), each experiment used a different HLA-mismatched human donor pair. Error bars in (b–e) represent Mean ± SEM. Error bars in (f, g) represent Mean ± SD. Log-rank (Mantel–Cox) test (a) and one-way ANOVA with Tukey’s multiple comparisons tests (b–e, g) were used. Source data are provided as a Source Data file.