title: Bacterial clearance in septic mice is modulated by MCP-1/CCL2 and nitric oxide.
abstract: Bacterial clearance is one of the most important beneficial consequences of the innate immune response. Chemokines are important mediators controlling leukocyte trafficking and activation, whereas reactive oxygen and nitrogen species are effectors in bacterial killing. In the present work, we used in vivo and in vitro models of infections to study the role of monocyte chemoattractant protein 1 (MCP-1)/CCL2 and nitric oxide (NO) in the bacterial clearance in sepsis. Our results show that MCP-1/CCL2 and NO levels are increased in the peritoneal cavity of mice 6 h after sepsis induced by cecal ligation and puncture. Pretreatment with anti-MCP-1/CCL2 monoclonal antibodies increased the number of colony-forming units (CFUs) recovered in the peritoneal lavage fluid. Moreover, CFU counts were increased in the peritoneal fluid of CCR2 mice subjected to cecal ligation and puncture. In vitro stimulation of peritoneal macrophages with recombinant MCP-1/CCL2 reduced CFU counts in the supernatant after challenge with Escherichia coli. Conversely, treatment with anti-MCP-1/CCL2 increased CFU counts under the same experimental condition. Stimulation of cultured macrophages with MCP-1/CCL2 and interferon had a synergistic effect on NO production. Macrophages from CCL2 mice showed a consistent decrease in NO production when compared with wild-type controls after stimulation with LPS + interferon. Finally, we showed incubation of macrophages with E. coli, and the ERK inhibitor U0126 increased CFU numbers and decreased intracellular levels of NO. In conclusion, we demonstrated for the first time that MCP-1/CCL2 has a crucial role in the clearance of bacteria by mechanisms involving increased expression of inducible NO synthase and production of NO by ERK signaling pathways.
authors.affiliation: Laboratório de Imunofarmacologia, Instituto Oswaldo Cruz, Instituto de Pesquisa Evandro Chagas, FIOCRUZ, Brazil.
authors.forename: Rachel N
authors.initials: RN
authors.lastname: Gomes
authors.forename: Mariana G A
authors.initials: MG
authors.lastname: Teixeira-Cunha
authors.forename: Rodrigo T
authors.initials: RT
authors.lastname: Figueiredo
authors.forename: Patricia E
authors.initials: PE
authors.lastname: Almeida
authors.forename: Silvio C
authors.initials: SC
authors.lastname: Alves
authors.forename: Patrícia T
authors.initials: PT
authors.lastname: Bozza
authors.forename: Fernando A
authors.initials: FA
authors.lastname: Bozza
authors.forename: Marcelo T
authors.initials: MT
authors.lastname: Bozza
authors.forename: Guy A
authors.initials: GA
authors.lastname: Zimmerman
authors.forename: Hugo C
authors.initials: HC
authors.lastname: Castro-Faria-Neto
mesh_terms: D000818:Animals; D001202:Ascitic Fluid / Q000382:microbiology; D001424:Bacterial Infections / Q000276:immunology* / Q000382:microbiology; D002070:Butadienes / Q000494:pharmacology; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000172:deficiency / Q000276:immunology*; D015169:Colony Count, Microbial; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000276:immunology; D020935:MAP Kinase Signaling System / Q000276:immunology; D008262:Macrophage Activation / Q000276:immunology; D017737:Macrophages, Peritoneal / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009569:Nitric Oxide / Q000096:biosynthesis / Q000276:immunology*; D009570:Nitriles / Q000494:pharmacology; D010587:Phagocytosis / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D018805:Sepsis / Q000276:immunology* / Q000382:microbiology
chemical_list: D002070:Butadienes; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D004791:Enzyme Inhibitors; D009570:Nitriles; D011994:Recombinant Proteins; C113580:U 0126; D009569:Nitric Oxide; D048049:Extracellular Signal-Regulated MAP Kinases