Toll-like receptor 5 agonist CBLB502 induces radioprotective effects in vitro

CBLB502 derived from Salmonella flagellin is a novel agonist of Toll-like receptor 5 （TLR5）. It has been shown that CBLB502 can exert high radioprotective efficacy on mice and primates from both GI and hematopoietic syndromes during whole-body irradiation with low toxicity and immunogen- icity. However, no effective system has been used to investigate the protective effect of CBLB502 against irradiation and the related mechanism in vitro. In this study, we investigated the radiopro- tective properties of CBLB502 in HEK293-N-T cells constitutively expressing human TLR5 and NF-KB-dependent luciferase. HEK293-N-T cells were treated with different doses of CBLB502 prior to 6~Co-~/ray irradiation. After irradiation, cell viability was real-time measured for 4 days by using the real-time cell analysis system. We found that CBLB502 was capable of efficiently maintaining the survival rate of irradiated HEK293-N-T cells. Then apoptotic cell death and cell cycle were detected by flow cytometry. The results showed that CBLB502 pre-treatment could reduce the apoptosis and promote the recovery of irradiated HEK293-N-T cells from G2-phase arrest in a dose-dependent manner. Our data indicated that CBLB502 has a direct radioprotective effect in vitro via anti-apoptosis and promotes cell cycle recovery. The method developed here could be an effective in vitro system to screen other TLR5-target radioprotectants like CBLB502.