ABSTRACT
Objective: To investigate the caspase-1 dependent inflammatory pathway activity and interleukin-1β (IL-1β) secretion in murine macrophage cell lines J774G8 infected with Leishmania major (L. major) using caspase-1 activity assay and ELISA. Methods: Novy-MacNeal-Nicolle biphasic medium was applied to produce promastigote form of L. major. Metacyclic promastigotes in the stationary phase were applied to infect macrophage. Caspase-1 activity and IL-1β secretion were assessed by the CPP32/caspase-1 fluorometric protease assay and ELISA IL-1β kits, respectively, with time intervals of 6, 18 and 30 h. Results: Our study showed an increase in caspase-1 activity and IL-1β secretion in infected samples compared to non-infected macrophages. The highest increase in IL-1β production was observed after 6 h of infection. Conclusions: These results arise that the activation of inflammasome pathway could be one of the innate immunity pathways against L. major.
ABSTRACT
OBJECTIVE@#To investigate the caspase-1 dependent inflammatory pathway activity and interleukin-1β (IL-1β) secretion in murine macrophage cell lines J774G8 infected with Leishmania major (L. major) using caspase-1 activity assay and ELISA.@*METHODS@#Novy-MacNeal-Nicolle biphasic medium was applied to produce promastigote form of L. major. Metacyclic promastigotes in the stationary phase were applied to infect macrophage. Caspase-1 activity and IL-1β secretion were assessed by the CPP32/caspase-1 fluorometric protease assay and ELISA IL-1β kits, respectively, with time intervals of 6, 18 and 30 h.@*RESULTS@#Our study showed an increase in caspase-1 activity and IL-1β secretion in infected samples compared to non-infected macrophages. The highest increase in IL-1β production was observed after 6 h of infection.@*CONCLUSIONS@#These results arise that the activation of inflammasome pathway could be one of the innate immunity pathways against L. major.