GvHD severity is reduced when recipients lack Asc or Nlrp3. (A and B) Asc^−/− or Asc^+/− C57BL/6 mice (n = 6 per group) underwent TBI followed by allo-HCT with C3H BM plus T cells. As control groups, WT C57BL/6 mice that received TBI alone (n = 6) or TBI plus C3H BM (n = 6) were used. Graphs show survival curves after transplantation (A) and histopathological scoring (apoptosis, inflammation) of the GvHD target organs (small intestine, large intestine and liver) on d10 after transplantation (B). The experiment was performed three times with similar results. One representative experiment is shown. (C–E) Nlrp3^−/− or Nlrp3^+/+ C57BL/6 mice (n = 10–12 per group) underwent TBI followed by allo-HCT with BALB/c BM alone or BALB/c BM plus T cells. T cell doses were 10⁵ (C), 10⁶ (D), or 3 × 10⁵ (E) as indicated. Graphs show survival curves after transplantation (C and D) and histopathological scoring (apoptosis, inflammation) of the GvHD target organs (small intestine, large intestine, and liver) on day 10 after transplantation (E). The experiment was performed twice, and the resulting survival data were pooled. (F) C57BL/6 mice underwent TBI, followed by transplantation with BALB/c BM alone (n = 10) or BALB/c BM plus T cells and were monitored for survival. Treatment was performed with PBS (n = 10), the Nlrp3 inhibitor glibenclamide (n = 16), or glibenclamide plus the TNF inhibitor Etanercept (n = 10) as indicated. Treatment with glibenclamide was started on day 0, and treatment with Etanercept was started on day 5. The experiment was performed twice, and the resulting data were pooled. (G) BALB/c mice were conditioned with FLU/CY based chemotherapy, followed by transplantation with C57/BL6 BM plus T cells, and then monitored for survival. Treatment was performed with vehicle (DMSO, n = 10), glibenclamide (n = 10), or Anakinra (n = 10). ***, P < 0.0001; **, P < 0.005; *, P < 0.05.