Flagella from Five Cronobacter Species Induce Pro-Inflammatory Cytokines in Macrophage Derivatives from Human Monocytes
Por:
Cruz-Córdova A., Rocha-Ramírez L.M., Ochoa S.A., Gónzalez-Pedrajo B., Espinosa N., Eslava C., Hernández-Chiñas U., Mendoza-Hernández G., Rodríguez-Leviz A., Valencia-Mayoral P., Sadowinski-Pine S., Hernández-Castro R., Estrada-García I., Munoz-Hernandez, O, Rosas I., Xicohtencatl-Cortes J.
Publicada:
21 dic 2012
Resumen:
Cronobacter spp. are opportunistic pathogens linked to lie-threatening infections in neonates and contaminated powdered infant formula that has been epidemiologically associated with these cases. Clinical symptoms of Cronobacter include necrotizing enterocolitis, bacteremia, and meningitis. Flagella from C. sakazakii are involved in biofilm formation and its adhesion to epithelial cells. We investigated the role of flagella from C. sakazakii ST1 and ST4, C. malonaticus, C. muytjensii, C. turicensis and C. dublinensis during the activation of cytokines (IL-8, TNF-a, and IL-10) in macrophage derivatives from human monocytes, which has not been extensively studied. The production and identity of flagella from the five Cronobacter species were visualized and recognized with anti-flagella antibodies by immunogold labeling through transmission electron microscopy. Purified flagella were dissociated into monomers in 12% SDS-PAGE Coomassie blue-stained gels showing a band of ~28 kDa and, in addition, mass spectrometry revealed the presence of several peptides that correspond to flagellin. Flagella (100 ng) induced the release of IL-8 (3314-6025 pg/ml), TNF-a (39-359 pg/ml), and IL-10 (2-96 pg/ml), in macrophage isolates from human monocytes and similar results were obtained when flagella were dissociated into monomers. Inhibition assays using three dilutions of anti-flagella antibodies (1:10, 1:100, and 1:200) suppressed the secretion of IL-8, TNF-a, and IL-10 between 95-100% using 100 ng of protein. A transfection assay using 293-hTLR5 cells showed IL-8 release of 197 pg/ml and suppression in the secretion of IL-8 when anti-hTLR5-IgA antibodies were used at different concentrations. These observations suggest that flagella and flagellin are involved in an inflammatory response dependent on TLR5 recognition, which could contribute to the pathogenesis of the bacteria. © 2012 Cruz-Córdova et al.
Filiaciones:
Cruz-Córdova A.:
Laboratorio de Bacteriología Intestinal, Departamento de Infectología, Hospital Infantil de México Federico Gómez, México D.F., Mexico
Rocha-Ramírez L.M.:
Laboratorio de Bacteriología Intestinal, Departamento de Infectología, Hospital Infantil de México Federico Gómez, México D.F., Mexico
Ochoa S.A.:
Laboratorio de Bacteriología Intestinal, Departamento de Infectología, Hospital Infantil de México Federico Gómez, México D.F., Mexico
Gónzalez-Pedrajo B.:
Univ Nacl Autonoma Mexico, Inst Fisiol Celular, Dept Genet Mol, Mexico City 04510, DF, Mexico
Espinosa N.:
Univ Nacl Autonoma Mexico, Inst Fisiol Celular, Dept Genet Mol, Mexico City 04510, DF, Mexico
Eslava C.:
Univ Nacl Autonoma Mexico, Fac Med, Dept Salud Publ, Mexico City 04510, DF, Mexico
Hernández-Chiñas U.:
Univ Nacl Autonoma Mexico, Fac Med, Dept Salud Publ, Mexico City 04510, DF, Mexico
Mendoza-Hernández G.:
Univ Nacl Autonoma Mexico, Fac Med, Dept Bioquim, Mexico City 04510, DF, Mexico
Rodríguez-Leviz A.:
Departamento de Patología, Hospital Infantil de México Federico Gómez, Dr. Márquez 162, Col. Doctores, Delegación Cuauhtémoc, México D.F., Mexico
Valencia-Mayoral P.:
Departamento de Patología, Hospital Infantil de México Federico Gómez, Dr. Márquez 162, Col. Doctores, Delegación Cuauhtémoc, México D.F., Mexico
Sadowinski-Pine S.:
Departamento de Patología, Hospital Infantil de México Federico Gómez, Dr. Márquez 162, Col. Doctores, Delegación Cuauhtémoc, México D.F., Mexico
Hernández-Castro R.:
Departamento de Ecología de Agentes Patógenos, Hospital General Dr. Manuel Gea González, Tlalpan, Mexico
Estrada-García I.:
Univ Nacl Autonoma Mexico, Ctr Ciencias Atmosfera, Mexico City 04510, DF, Mexico
Rosas I.:
Departamento de Inmunología Molecular, Escuela Nacional de Ciencias Biológicas, IPN. Unidad Profesional Prolongación, Miguel Hidalgo, D. F., Mexico
Xicohtencatl-Cortes J.:
Laboratorio de Bacteriología Intestinal, Departamento de Infectología, Hospital Infantil de México Federico Gómez, México D.F., Mexico
Gold
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