Fig. 3. CD45RA/CD276 depletion significantly delayed the onset of GVHD symptoms and remarkably alleviated their severity, leading to the rescue of NSG-Ab°DR4 mice from GVHD mortality in HLA-DR-mismatched setting.

NSG-Ab°DR4 mice received one of the three different grafts: HLA-DR4^negative or HLA-DR4^positive donor’s bulk CD4⁺ T cells (black), or CD45RA-depleted memory CD4⁺ T cells (blue), or CD45RA/CD276-depleted memory CD4⁺ T cells (pink). Mice were scored daily individually for signs of GVHD. a Survival, mean + SD of total GVHD scores in each cohort, b total scores or respective symptom score before euthanization, c schedule of onset of GVHD (days), and d the maximum frequency of human T cells among whole PBMCs. (n = 6 (bulk CD4⁺ T cells), n = 5 (memory CD4⁺ T cells, CD276-depleted memory CD4⁺ T cells)). One mouse in the cohort of memory CD4⁺ T-cell transplantation was excluded from analysis due to sacrification for non-GVHD related complication (uterus prolapse). Shown are the representative data. We performed preliminary experiments at least twice by using the identical donors with consistent results. The Kaplan–Meier method was used to assess survival rate using the log-rank test (a). One-way ANOVA followed by Tukey’s multiple comparison test was used (b, c). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.