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Analysis of inflammatory cytokine expression in the urinary tract of BALB/c mice infected with Proteus (P.) mirabilis and enteroaggregative Escherichia (E.) coli (EAEC) strains

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Abstract

This study aimed to analyze the proinflammatory cytokine mRNA expression in the urinary tract of BALB/c mice infected with bacterial strains with uropathogenic potential. Groups of four 6-week-old female BALB/c mice were intraurethrally inoculated with 5 × 107 colony-forming units (CFU) of P. mirabilis ATCC29906, EAEC O42, P. mirabilis RTX339, or sterile saline (control group) and then sacrificed at 0, 2, 4, 7, or 10 days post-infection (p.i.). Samples were cultured to determine the CFU/mL in urine or CFU/g in the bladders and kidneys. Cytokine expression (tumor necrosis factor (TNF)-α and interleukin (IL)-1β, -6, and -8) was evaluated in the target organs using real-time PCR and immunohistochemistry; histology was examined with hematoxylin and eosin staining. The results are presented as the means and standard deviations and were compared using one-way ANOVA, with p < 0.05 indicating significant differences. Bacteriuria was not detected in the infected groups; bacterial colonization occurred in the target organs at all time points, but was higher in mice infected with EAEC O42 or P. mirabilis RTX339 at 7 days p.i. The expression of all cytokine mRNAs was seen, but only the levels of the IL-8 protein increased in situ at 7 days p.i. in the P. mirabilis RTX339 and EAEC O42 groups in both organs. Morphological alterations, observed in all of the infected groups, were more prominent in the EAEC O42 and P. mirabilis RTX339 groups. The findings provide insights into the uropathogenicity and inflammatory cytokine expression in the urinary tract of mice infected with three previously untested bacterial strains.

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Acknowledgements

The authors thank Dr. Julia Perez Ramos (Lab Biologia Experimental UAMX) and Dr. Osvaldo Lopez (Lab Histopatologia UAMX) for allowing us to use a rotor-Gene 6000 and microtome, respectively.

Funding

Araceli Melendez Avalos (Fellowship No. 600558) was provided financial support by the Consejo Nacional de Ciencia y Tecnologia (CONACyT) during her studies in the program of Maestria en Ciencias Farmaceuticas (MCF), Universidad Autonoma Metropolitana Unidad Xochimilco (UAMX).

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Authors and Affiliations

  1. Maestria en Ciencias Farmaceuticas, Universidad Autonoma Metropolitana Unidad Xochimilco, Calzada del Hueso No. 1100, CP 04960, CDMX, Mexico

    Araceli Melendez-Avalos

  2. Departamento de Sistemas Biologicos, Universidad Autonoma Metropolitana Unidad Xochimilco, Calzada del Hueso No. 1100, CP 04960, CDMX, Mexico

    Teresita Sainz-Espuñes, Laura Estela Castrillón-Rivera, Felipe Mendoza-Pérez, Alejandro Palma-Ramos, Jorge Ismael Castañeda-Sánchez & Elisa Maria Drago-Serrano

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  1. Araceli Melendez-Avalos
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Contributions

Melendez-Avalos Araceli: performed the experiments, analyzed the data, and designed the figures.

Sainz Espuñes Teresita: conceived the study.

Castrillón-Rivera Laura Estela: contributed to the bacterial count assays.

Mendoza-Pérez Felipe: provided advice on the experiments and processed the data.

Palma-Ramos Alejandro: provided advice on the immunohistochemistry assay.

Castañeda-Sánchez Jorge Ismael: primer design and provided advice on the RT-PCR assay.

Drago-Serrano Maria Elisa: wrote the paper.

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Correspondence to Elisa Maria Drago-Serrano.

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Melendez-Avalos, A., Sainz-Espuñes, T., Castrillón-Rivera, L.E. et al. Analysis of inflammatory cytokine expression in the urinary tract of BALB/c mice infected with Proteus (P.) mirabilis and enteroaggregative Escherichia (E.) coli (EAEC) strains. Folia Microbiol 65, 133–142 (2020). https://doi.org/10.1007/s12223-019-00714-2

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