Present study was undertaken to evaluate radioprotective and immunomodulatory activities of a novel semiquinone glucoside derivative (SQGD) isolated from Bacillus sp. INM-1 in C57 BL/6 mice. Whole body survival study was performed to evaluate in vivo radioprotective efficacy of SQGD. To observe effect of SQGD on immunostimulation, Circulatory cytokine (i.e., interleukin-2 (IL-2), IFN-γ, IL-10, granulocyte colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), and macrophage colony stimulating factor (M-CSF) expression was analyzed in serum of irradiated and SQGD treated mice at different time intervals using ELISA assay. Results of the present investigation indicated that SQGD pre-treatment (-2 h) to lethally irradiated mice provide ∼ 83% whole body survival compared with irradiated mice where no survival was observed at 30(th) post irradiation day. Significant (p < 0.05) induction in IL-2 and IFN-γ expression was observed at all tested time intervals with SQGD pre-treated irradiated mice as compared with irradiated mice alone. However, sharp increase in IL-10 expression was observed in irradiated mice which were found to be subsidized in irradiated mice pre-treated with SQGD. Similarly, significant (p < 0.05%) induction in G-CSF, M-CSF and GM-CSF expression was observed in irradiated mice treated with SQGD as compared with irradiated control mice at tested time intervals. In conclusion, SQGD pre-treatment to irradiated mice enhanced expression of IL-12 and IFN-γ while down-regulated IL-10 expression and thus modulates cytoprotective pro-inflammatory TH1 type immune response in irradiated mice. Further, SQGD pre-treatment to irradiated mice accelerate G-CSF, GM-CSF and M-CSF expression suggesting improved haematopoiesis and enhanced cellular immune response in immuno-compromised irradiated mice that may contribute to in vivo radiation protection.