High-dose XRT followed by adjuvant α-OX40 most effectively suppressed local tumor growth and reduced lung metastases. A, Mice injected in the right hind legs with anti-PD1-resistant 344SQ tumors (n = 6 per group) were treated with intratumoral α-OX40 (three 200-μg injections) before XRT (induction); during XRT (concurrent); or after XRT (adjuvant). Tumor growth volumes were plotted. 12Gyx3 + α-OX40 was superior to 12Gyx2 + α-OX40 (Fig. 1C) (two-way ANOVA, P=0.0265); 12Gyx3 vs. 12Gyx3 + adjuvant α-OX40 (P= 0.0114 for the last point on day 25); α-OX40 vs. 12Gyx3 + adjuvant α-OX40 (two-way ANOVA, P=0.0005). B, Lungs from mice in 2A were harvested, stained with Bouin’s fixative solution, and numbers of metastases counted. *P ≤ 0.05.