Fig. 5. Blockade of Aurora kinase A and JAK2 reduces xenogeneic GVHD and preserves the in vivo generation of potent antitumor CTL.

NSG mice received human PBMCs (30 × 10⁶ cells) by intraperitoneal injection, with alisertib (30 mg/kg daily), TG101348 (45 mg/kg twice a day), a combination of alisertib and TG101348, or vehicle administered by oral gavage from day 0 to day +14. (A) Percent survival is shown among the four groups (log-rank test). (B) Graph shows mean GVHD clinical scores ± SEM for each group of mice (P = 0.02 at day +30, vehicle versus combo, Mann-Whitney). Pooled data are from two independent experiments. n = 7 to 8 mice per group. NSG mice were transplanted with human PBMCs as described, with AJI-100 (50 mg/kg daily) or vehicle administered ip from day 0 to day +14. (C and D) Percent survival (log-rank test) and mean GVHD clinical scores ± SEM (Mann-Whitney) are demonstrated. Pooled data are from two independent experiments. n = 8 mice per group. (E) Representative contour plots show expression of H3Ser¹⁰ and STAT3 phosphorylation among human CD3⁺ T cells harvested from recipient spleens at day +14. (F) Bar graph shows the mean % pH3Ser¹⁰⁺ and % pSTAT3⁺ T cells ± SEM among AJI-100–treated and vehicle-treated mice at day +14 (n = 6 mice per group, two independent experiments, Mann-Whitney). (G) Graph depicts mean specific lysis ± SD by human CD8⁺ CTL generated in vivo using NSG mice transplanted with human PBMCs and vaccinated with irradiated U937cells (1 × 10⁷) on days 0 and +7. Results shown are from one of two independent experiments, using a total of seven mice per group. U937 lysis was measured by released fluorescence after 4 hours (vehicle versus AJI-100, not significant, Mann-Whitney). *P < 0.05, **P = 0.001 to 0.01, ***P = 0.0001 to 0.001, ****P < 0.0001.