Genome Sequencing and Assembly of Enterotoxigenic Escherichia coli E9034A: Role of LngA, CstH, and FliC in Intestinal Cell Colonization and the Release of the Proinflammatory Cytokine IL-8
Por:
Rodríguez-Martínez R., Ochoa S.A., Valle-Rios R., Jaimes-Ortega G.A., Hernández-Castro R., Mancilla-Rojano J., Castro-Escarpulli G., López-Saucedo C., Estrada-García T., Cruz-Córdova A., Xicohtencatl-Cortes J.
Publicada:
1 ene 2025
Resumen:
Enterotoxigenic Escherichia coli (ETEC) produces two types of enterotoxins, LTs and STs, as well as several colonization factors (CFs), including CS21, CS3 fimbriae, and flagellar structures. This study investigated how these structures contribute to ETEC colonization and the immune response in HT-29 and HuTu-80 intestinal cells. ETEC strains with single, double, and triple mutations in the lngA, cstH, and fliC genes were generated and confirmed using PCR and Western blotting. The colonization of HT-29 and HuTu-80 intestinal cells by the ETEC E9034A strain, which was fully sequenced using a hybrid approach involving both Illumina and Oxford Nanopore technologies, was used to generate the mutant and recombinant proteins. The colonization and adherence of E9034A and its mutants were assessed through colony-forming unit (CFU) counts. Cytokine levels were assessed using flow cytometry and analyzed via FlowJo 7.6.1. Quantitative analysis revealed that the absence of the lngA, cstH, and fliC genes significantly (p < 0.01) reduced ETEC adherence to HT-29 and HutU-80 cells. In addition, only ETEC strains expressing the FliC protein induced IL-8 secretion. These findings suggest that LngA, CstH, and FliC in ETEC E9034A enhance adherence to intestinal cells and trigger the release of IL-8. © 2025 by the authors.
Filiaciones:
Rodríguez-Martínez R.:
Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, 11340, Mexico
Laboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Mexico City, 06720, Mexico
Ochoa S.A.:
Laboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Mexico City, 06720, Mexico
Valle-Rios R.:
Unidad Universitaria de Investigación en Cáncer e Inmunología, División de Investigación, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City, 04510, Mexico
Unidad de Investigación en Inmunología y Proteómica, Hospital Infantil de México Federico Gómez, Mexico City, 06720, Mexico
Jaimes-Ortega G.A.:
Unidad Universitaria de Investigación en Cáncer e Inmunología, División de Investigación, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City, 04510, Mexico
Unidad de Investigación en Inmunología y Proteómica, Hospital Infantil de México Federico Gómez, Mexico City, 06720, Mexico
Posgrado en Biología Experimental, Departamento de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana, Mexico City, 09310, Mexico
Hernández-Castro R.:
Departamento de Ecología de Agentes Patógenos, Hospital General Dr. Manuel Gea González, Mexico City, 14080, Mexico
Mancilla-Rojano J.:
Laboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Mexico City, 06720, Mexico
Posgrado en Ciencias Biológicas, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City, 04510, Mexico
Castro-Escarpulli G.:
Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City, 11340, Mexico
López-Saucedo C.:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados CINVESTAV-IPN, Mexico City, 07360, Mexico
Estrada-García T.:
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados CINVESTAV-IPN, Mexico City, 07360, Mexico
Cruz-Córdova A.:
Laboratorio de Investigación en Inmunoquímica, Hospital Infantil de México Federico Gómez, Mexico City, 06720, Mexico
Xicohtencatl-Cortes J.:
Laboratorio de Investigación en Bacteriología Intestinal, Hospital Infantil de México Federico Gómez, Mexico City, 06720, Mexico
gold, All Open Access; Gold Open Access
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