Figure 1.

HMGB1 Is Released In Vitro and In Vivo during oHSV Tumor Therapy

(A) HMGB1 levels in conditioned medium from HSVQ-infected U87 glioblastoma cells (MOI = 0.1 or MOI = 1) and U87 cells infected with HSVQ and co-cultured with HUVEC cells (U87 + HUVEC). Conditioned medium was harvested 24 hr after treatment and HMGB1 levels measured by ELISA. Data shown are mean HMGB1 levels measured by ELISA in the conditioned medium ± SD. (B) HMGB1 released in culture medium of the indicated cells after infection with HSVQ with or without HUVEC cell overlay as indicated (MOI = 0.1). HMGB1 secretion in the cells co-cultured with HUVEC was analyzed by ELISA. Data shown are mean HMGB1 levels measured by ELISA in the conditioned medium ± SD. (C) HUVEC or U251 cells were transfected with either scrambled control or HMGB1 targeting siRNA. Control or HMGB1 siRNA-transfected glioma cells were infected with HSVQ (MOI = 0.01), and then the cells were overlayed with endothelial cells transfected with either scrambled control or HMGB1-targeting siRNA. 24 hr after treatment with HSVQ or no virus and HMGB1, levels were measured by ELISA. Data shown are mean HMGB1 levels measured by ELISA in the conditioned medium ± SD. (D) Mice bearing intracranial GBM tumors were treated with HSVQ intratumorally (5 × 10⁵ pfu/mouse). Five days later, serum was harvested and HMGB1 levels were measured by ELISA (n = 5/group; NI, non-injected; PBS, saline-injected; oHSV, HSQ-injected animals). *p < 0.05; **p < 0.01.