Effects of Clostridium septicum infection on macrophages cells line RAW264.7

VILLA MARÍA CECILIA; CÁCERES CLAUDIA SOLEDAD; GALLO GIOVANNA LUCRECIA; CASTRO MELISA; CORTIÑAS TERESA INÉS; SOTOMAYOR CLAUDIA ELENA; STEFANINI ANA MARÍA

Mendoza

Congreso; XXIV Reunión Científica Anual de La Sociedad de Biología de Cuyo.; 2008

Sociedad de Biología de Cuyo

Clostridum septicum is a pathogenic anaerobe microorganism that causes traumatic and non traumatic gas gangrene in most animals and man. The disease have a rapid course and is always fatal. For the establishment of infection pathogens have to overcome or evade host defenses so the response of macrophages, key cells of the immune system, to C. septicum is necessary to understand its pathogenesis. The aim of this study was to determine the effects of C. septicum infection to mouse monocytic-macrophage cell line RAW 264.7.The cell line was cultured in Dulbecco´s modified Eagle´s medium containing 10% fetal bovine serum. Live, death and antibody opsonized cells of C. septicum ATCC 12464, collected at the logarithmic phase of grow in a clostridial medium under anaerobic conditions, were used to infect RAW cells at different multiplicity of infection (moi) and time of post-infection. The viability, phagocytic activity and nitric oxide (NO) production was determine using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), nitro blue tetrazolium (NBT) assays and Griess reaction respectively. The results showed a lost of viability depending on the moi and time assayed. Live cells induced the highest phagocytic activity and NO was sustained at lower moi 24 h post infection. The cell line RAW264.7 can be successfully used for in vitro studies of C. septicum infection.