Effects of NF-kappa B1 (p50) targeted gene disruption on ionizing radiation-induced NF-kappa-B activation and TNF alpha, IL-1 alpha, IL-1 beta and IL-6 mRNA expression in vivo

Purpose: To investigate the role of the NF-kappaB] (p50) gene in ionizing r adiation (IR)-induced NF-kappaB activation and TNF alpha, IL- 1 alpha, IL-1 beta and IL-6 mRNA expression in vivo. Materials and methods: NF-kappaB activation was analysed by the gel shift/s upershift assay and the levels or TNF alpha, IL-1 alpha, IL-1 beta and IL-6 mRNA were measured using RNase protection assay (RPA). Various tissues fro m BALB/c, B6,129P-Nfkb1 (NF-kappa B1 or p50 gene knockout, p50(-/-)) and B6 ,129PF2 (wild-type, p50(+/+)) mice were analysed before or after exposure t o a lethal dose (8.5 Gy) of total-body gamma -irradiation. Results: Exposure of BALB/c mice to total-body IR selectively activated NF- kappaB in the spleen, mesenteric lymph nodes (LN) and bone marrow (BM). Gel supershift assay using polyclonal antibodies against NF-kappaB p50, p65 or c-Rel protein revealed that the NF-kappaB p50 subunit is a critical compon ent of the NF-kappaB complexes activated by IR in vivo. Discretely augmente d TNF alpha, IL-1 alpha, IL-1 beta and IL-6 mRNA expression was found in th e spleen, LN and BM after BALB/c mice received IR. However, mice lacking th e p50 gene (p50(-/-)) showed a significant reduction in IR-induced activati on of NF-kappaB and increases in TNF alpha, IL-1 alpha IL-1 beta and IL-6 m RNA expression, as compared with that of wild-type mice (p50(+/+)). Conclusions:The NF-kappaB p50 subunit is a critical component of the NF-kap paB complexes activated by IR and it plays an important role in mediating I R-induced TNF alpha, IL-1 alpha, IL-1 beta and IL-6 mRNA expression in vivo .