N-acetyl tryptophan glucopyranoside (NATG) as a countermeasure against gamma radiation-induced immunosuppression in murine macrophage J774A.1 cells.
Radiation exposure to immune system induces imbalance in cytokines expression involved in Th1/Th2 homeostasis perturbations. In the present study, N-acetyl tryptophan glucoside (NATG), a bacterial secondary metabolite, was evaluated for its possible radioprotective potential to immune system using J774A.1 murine macrophages. In this study, expression of IFN-γ, TNF-α, IL-10, IL-2, IL-12, IL-13 and IL-17A cytokines was analyzed in irradiated and NATG pretreated cells using ELISA assay. Results of the study indicated that irradiated macrophages (NK-1R+cells) pretreated with NATG showed higher (p < .05) survival at all observed time-intervals (2 h-48 h) as compared to irradiated (20Gy) cells that were not pretreated with NATG. However, NATG pretreatment to irradiated HEK293T cells (that did not express NK-1Receptor) did not provide significant survival, suggesting NK-1R involvement in NATG-mediated radioprotection. Cytokine expression analysis demonstrated that NATG pre-treated plus irradiated J774A.1 murine macrophages exhibited increased IFN-γ levels (∼90%) with significant decrease in TNF-α at 24h as compared to irradiated cells. Further, significant decrease (∼20%) in IL-10 and IL-2 (∼26%) levels was observed in irradiated macrophages pretreated with NATG as compared to only irradiated cells. A sharp improvement in IL-17A (∼92%) and IL-12 (∼116%) expression was observed in irradiated macrophages pretreated with NATG as compared to only irradiated cells. Hence, NATG pre-treatment to irradiated macrophages induced IFN-γ, IL-17A and IL-12 expression, but suppresses TNF-α, IL-10 and IL-2 expressions. Conclusively, NATG pretreatment overcomes radiation-induced Th2 immune response by improving Th1 responsive cytoprotective cytokines IFN-γ, IL-17A and IL-12 in irradiated macrophages possibly by NK-1R antagonistic mechanism, and thus contributes to radioprotection.
a Radiation Biotechnology Group, Division of Radiation Biosciences , Institute of Nuclear Medicine and Allied Sciences , Delhi, India.
Poonam
P
Malhotra
a Radiation Biotechnology Group, Division of Radiation Biosciences , Institute of Nuclear Medicine and Allied Sciences , Delhi, India.
Manish
M
Adhikari
a Radiation Biotechnology Group, Division of Radiation Biosciences , Institute of Nuclear Medicine and Allied Sciences , Delhi, India.
Saurabh
S
Mishra
a Radiation Biotechnology Group, Division of Radiation Biosciences , Institute of Nuclear Medicine and Allied Sciences , Delhi, India.
Sumit
S
Singh
b School of Biotechnology, Gautam Buddha University , Greater Noida , Gautam Buddh Nagar , Uttar Pradesh , India.
Piyush
P
Kumar
a Radiation Biotechnology Group, Division of Radiation Biosciences , Institute of Nuclear Medicine and Allied Sciences , Delhi, India.
Shravan Kumar
SK
Singh
a Radiation Biotechnology Group, Division of Radiation Biosciences , Institute of Nuclear Medicine and Allied Sciences , Delhi, India.
Raj
R
Kumar
D000818:Animals; D016207:Cytokines / Q000378:metabolism*; D005720:Gamma Rays; D057809:HEK293 Cells; D006801:Humans; D007378:Interleukins / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D011839:Radiation, Ionizing; D014364:Tryptophan / Q000378:metabolism*
D016207:Cytokines; D007378:Interleukins; D014364:Tryptophan
Ionizing radiation; Th1/Th2 homeostasis; bacterial metabolite; cytokines; radioprotection