Figure 6.

CRBN controls T_E phenotype and harnesses GVHD pathogenesis. (A) Schematic of GVHD experiments. (B) Effects of GVHD in a model of BALB/c mice receiving C57Bl/6 T-cell–depleted bone marrow, with and without Crbn^+/+ or Crbn^−/− T cells (H2^b) after lethal body irradiation. The percentage of body weight (top) and survival (bottom) of recipient BALB/c H2^d mice was monitored at the indicated intervals. (C) The percentage of IFN-γ–producing CD4⁺ and CD8⁺ T cells in donor Crbn^+/+ or Crbn^−/− T cells derived from the GVHD model after restimulation with phorbol myristate acetate and ionomycin ex vivo on day 14 after transplantation. (D) pERK1/2, total ERK1/2, and Myc protein levels, as determined by western blot analysis in Crbn^+/+ and Crbn^−/− CD8⁺ T cells, with or without 25 μM PD98059 24 hours after activation with anti-CD3ε+anti-CD28. (E) qRT-PCR analysis of Myc, Tbx21, Eomes, Ifng, and Gzmb mRNA levels in Crbn^+/+ vs Crbn^−/− CD8⁺ T cells 24 hours after activation with anti-CD3ε+anti-CD28^+/− cotreatment with 25 μM PD98059. All results are representative of at least 2 independent experiments. n.s., not significant; *P < .05; **P < .01; ****P < .0001.