Salmonella-induced changes in the gut microbiota and immune response genes transcriptome during administration of vancomycin and Bacteroides fragilis

Authors

  • Yu. V. Bukina Zaporizhzhia State Medical University,
  • A. M. Kamyshnyi Zaporizhzhia State Medical University,
  • N. N. Polishchuk Zaporizhzhia State Medical University,
  • I. A. Topol Zaporizhzhia State Medical University,

DOI:

https://doi.org/10.14739/2310-1237.2017.1.97504

Keywords:

microbiome, vancomycin, Salmonella, Bacteroides, Real-Time PCR

Abstract

The aim. To study Salmonella-induced changes in the intestinal wall microbiota, the expression of Salmonella effector proteins SipA, SopB, SopE2 and transcriptional activity of genes FFAR2, Foxp3, RORγt in rat GALT during administration of vancomycin and B.fragilis.

Methods. Investigations of qualitative and quantitative composition of the microbiota of the wall of the small intestine were carried out, and the expression level of rat genes Foxp3, Rorc (Royt), FFAR2 and Salmonella effector proteins SipA, SopB and SopE2 were determined by RT-PCR, the relationship between groups of microorganisms was established.

Results. Administration of B.fragilis against the background pre-treatment with vancomycin and Salmonella infection alters the quantitative composition of the microbiota in the wall of the small intestine contents: a decrease in Salmonella spp., E.coli, P.aeruginosa, Proteus spp., Enterobacter spp., Klebsiella spp. and Shigella spp., as well as increasing Bacteroides spp., E.faecalis, E.faecium and Peptostreptococcus anaerobius. The level of expression of Salmonella effector proteins in animals with the combined administration of vancomycin and S.enteritidis (I group), S.typhimurium (II group) increased: SopB – 101 and 20 times; SopE2 - 80 and 2 times; SipA - 613 times (II group), and also 5-fold decrease was noted in the I group. Relative normalized number of mRNA of genes FFAR2, Foxp3, RORγt in GALT of rats in groups III and IV increased: FFAR2 - 2.7 and 5.4 times; Foxp3 - 2.5 and 85 times, RORγt level decreased by 70% and only in IV group.

Conclusions. Using B.fragilis creates conditions for the correction of Salmonella-induced changes of the intestinal microbiome. Pretreatment of animals with vancomycin causes increased transcriptional activity of genes SipA, SopB and SopE2, except SipA after administration of S.enteritidis. Administration of B.fragilis increases the level of mRNA of genes FFAR2 and Foxp3 in GALT and reduces RORγt after infection with S. typhimurium.

References

Purchiaroni, F., Tortora, A., Gabrielli, M., Bertucci, F., Gigante, G., Ianiro, G., et al. (2013) Role of the intestinal microbiota and the immune system. Medical Pharmacology Science, 17, 323–333.

Kamada, N., Seo, S-U., Chen, G. Y., & Núñez, G. (2013) Role of the gut microbiota in immunity and inflammatory disease. Nature Reviews Immunoljgy, 13, 321–335. doi:10.1038/nri3430.

Bukina, Y. V., Kamyshny, A. M., & Polishchuck, N. N. (2016) Оpredelenie spektra genov rezistentnosti k antibiotikam u fenotipicheski rezistentnykh shtammov pristenochnoj kishechnoj mikrobioty u krys metodom PCR-RV [Determination of the spectrum of antibiotic resistance genes have phenotypic resistant strains of parietal intestinal microbiota in rats by RT-PCR]. Annaly Mechnikovskogo instituta, 2, 21–27. [in Russian].

Bukina, Yu. V., Kamyshnyi, O. M., & Polishchuck, N. M. (2016) Vyznachennia kilkisnoho skladu mikrobioty u prystinkovomu vmisti kyshkivnyka u shchuriv metodom PLR-RCh [Determination quantitative composition of the microbiota in parietal intestinal surface in rats by PCR real-time]. Infektsiini khvoroby, 3, 78–81. [in Ukrainian].

Agbor, T. A., & McCormick, B. A. (2011) Salmonella effectors: important players modulating host cell function during infection. Cell Microbiology, 13, 1858–69. doi: 10.1111/j.1462-5822.2011.01701.x.

Behnsen, J., Perez-Lopez, A., Nuccio, S. P., & Raffatellu, M. (2015) Exploiting host immunity: the Salmonella paradigm. Trends Immunology, 36, 112–20. doi: 10.1016/j.it.2014.12.003.

Keestra-Gounder, A. M., Tsolis, R. M., & Bäumler, A. J. (2015) Now you see me, now you don't: the interaction of Salmonella with innate immune receptors. Natura Review Microbiology, 13, 206–16. doi: 10.1038/nrmicro3428.

Blacher, E., Levy, M., Tatirovsky, E., & Elinav, E. (2017) Microbiome-Modulated Metabolites at the Interface of Host Immunity. Immunology, 15, 572–580. doi: 10.4049/jimmunol.1601247.

Kim, M. H., Kang, S. G., Park, J. H., Yanagisawa, M., & Kim, C. H. (2013) Short-chain fatty acids activate GPR41 and GPR43 on intestinal epithelial cells to promote inflammatory responses in mice. Gastroenterology, 145, 396–410. doi: 10.1053/j.gastro.2013.04.056.

Smith, P. M., Howitt, M. R., Panikov, N., Michaud, M., Gallini, C. A., Bohlooly, Y. M., et al. (2013) The microbial metabolites, short-chain fatty acids, regulate colonic Treg cell homeostasis. Science, 341, 569–573. doi: 10.1126/science.1241165.

Sheikh, A., Charles, R. C., Sharmeen, N., Rollins, S. M., Harris, J. B., Bhuiyan, M. S., et al. (2011) In Vivo Expression of Salmonella enterica Serotype Typhi Genes in the Blood of Patients with Typhoid Fever in Bangladesh. PLoS Neglected Tropical Diseases, 5, e1419. doi: 10.1371/journal.pntd.0001419.

Ma, J., Zhang, Y. G., Xia, Y., & Sun, J. (2010) The inflammatory cytokine tumor necrosis factor modulates the expression of Salmonella typhimurium effector proteins. Inflammation, 12, 42. doi: 10.1186/1476-9255-7-42.

Que, F., Wu, S., & Huang, R. (2013) Salmonella pathogenicity island 1 (SPI-1) at work. Curr Microbiology, 66, 582–7. doi: 10.1007/s00284-013-0307-8.

LaRock, D. L., Chaudhary, A., & Miller, S. I. (2015) Salmonellae interactions with host processes. Nature Review Microbiology, 13, 191–205. doi: 10.1038/nrmicro3420.

Keestra, A. M., & Bäumler, A. J. (2014) Detection of enteric pathogens by the nodosome. Trends Immunology, 35, 123–130. doi: 10.1016/j.it.2013.10.009.

Keestra, A. M., Winter, M. G., Auburger, J. J., Frässle, S. P., Xavier, M. N., Winter, S. E., et al. (2013) Manipulation of small Rho GTPases is a pathogen-induced process detected by NOD1. Nature, 496, 233–237. doi: 10.1038/nature12025.

Narayanan, L. A., & Edelmann, M. J. (2014) Ubiquitination as an efficient molecular strategy employed in salmonella infection. Frontiers Immunology, 25, 558. doi: 10.3389/fimmu.2014.00558.

Hapfelmeier, S. (2004) Role of the Salmonella pathogenicity island 1 effector proteins SipA, SopB, SopE, and SopE2 in Salmonella enterica subspecies 1 serovar Typhimurium colitis in streptomycin-pretreated mice. Infection Immunology, 72, 795–809.

Miki, T., Goto, R., Fujimoto, M., Okada, N., & Hardt, W. D. (2017) The Bactericidal Lectin RegIIIβ Prolongs Gut Colonization and Enteropathy in the Streptomycin Mouse Model for Salmonella Diarrhea. Cell Host Microbe, 21(2), 130–131.

Corrêa-Oliveira, R., Fachi, J. L., Vieira, A., Sato, F. T., & Vinolo, M. A. (2016) Regulation of immune cell function by short-chain fatty acids. Clinical & Translational Immunology, 22, 73. doi: 10.1038/cti.2016.17.

Maslowski, K. M., Vieira, A. T., Ng, A., Kranich, J., Sierro, F., Yu, D., et al. (2009) Regulation of inflammatory responses by gut microbiota and chemoattractant receptor GPR43. Natur, 29, 1282–6. doi: 10.1038/nature08530.

Sun, M., Wu, W., Liu, Z., & Cong, Y. (2017) Microbiota metabolite short chain fatty acids, GPCR, and inflammatory bowel diseases. Gastroenterology, 52, 1–8. doi:10.1007/s00535-016-1242-9.

Surana, N. K., & Kasper, D. L. (2012) The yin yang of bacterial polysaccharides: lessons learned from B. fragilis PSA. Immunology Review, 245, 13–26. doi: 10.1111/j.1600-065X.2011.01075.x.

Zeng, H., & Chi, H. (2015) Metabolic control of regulatory T cell development and function. Trends Immunology, 36, 3–12. doi: 10.1016/j.it.2014.08.003

How to Cite

1.
Bukina YV, Kamyshnyi AM, Polishchuk NN, Topol IA. Salmonella-induced changes in the gut microbiota and immune response genes transcriptome during administration of vancomycin and Bacteroides fragilis. Pathologia [Internet]. 2017Apr.7 [cited 2024Nov.2];(1). Available from: http://pat.zsmu.edu.ua/article/view/97504

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Section

Original research