Mohammad Reza Mahmoudian Sani1, Mehrdad Moosazadeh Moghaddam2, Hossein Aghamollaei3, Kazem Hassanpour4, Ramezan Ali Taheri5, Gholamreza Farnoosh6. 1. Department of Molecular Medicine & Genetic, Hamedan University of Medical Sciences, Hamedan, Iran. 2. Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. 3. Young Researchers and Elites Club, North Tehran Branch, Islamic Azad University, Tehran, Iran. 4. Medical School, Sabzevar University of Medical Sciences, Sabzevar, Iran. 5. Nanobiotechnology Research Centre, Baqiyatallah University of Medical Sciences, Tehran, Iran. 6. Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. Electronic address: rzfarnoosh@yahoo.com.
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.
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.
Authors: Renan V H de Carvalho; Warrison A Andrade; Djalma S Lima-Junior; Marisa Dilucca; Caroline V de Oliveira; Kun Wang; Paula M Nogueira; Jeronimo N Rugani; Rodrigo P Soares; Stephen M Beverley; Feng Shao; Dario S Zamboni Journal: Cell Rep Date: 2019-01-08 Impact factor: 9.423