The effect of immunoregulatory bacteria on the transcriptional activity of Foxp3 and RORyt genes in the gut-associated lymphoid tissue with Salmonella-induced inflammation in the presence of vancomycin and Ваcteroides fragilis
-
Yuliia Bukina
,
Marina Thyhonovska
,
Mariya Koval
,
Mariya Marushchak
,
Inna Krynytska
,
Aleksandr Kamyshnyi
Abstract
Background and Objectives: Intestinal microbiota is involved in the development and maintenance of immune homeostasis. This study was conducted to investigate the levels of key immunoregulatory bacteria in the intestinal wall-associated microflora and its effect on the transcriptional activity of the Foxp3 and RORyt genes in the gut-associated lymphoid tissue (GALT) of rats with Salmonella-induced inflammation, both untreated and treated with vancomycin and Bacteroides fragilis.
Materials and Methods: To determine the levels of immunoregulatory bacteria in GALT of rats Q-PCR was used to identify them by species-specific 16S rDNA genes. Transcriptional activity of Foxp3 and RORyt genes was determined using Q-PCR with reverse transcription.
Results: In animals treated with both vancomycin and Salmonella, the levels of segmented filamentous bacteria (SFB) increased while Akkermansia muciniphila and Faecalibacterium prausnitzii decreased. In rats that received pretreatment with vancomycin and then were infected with S. Enteritidis and S. Typhimurium, the levels of SFB increased, and the number of Bacteroides-Prevotela group, A. muciniphila, Clostridium spp. clusters XIV, IV, and F. prausnitzii significantly decreased, decreasing Foxp3 and increasing Rorγt mRNA expression. Administration of B. fragilis to animals treated with S. Enteritidis or S. Typhimurium and pre-treated with vancomycin caused a decrease in SFB and Rorγt mRNA levels and conversely, increased the numbers of the Bacteroides-Prevotela group, Clostridium spp. clusters XIV, IV, A. muciniphila, F. prausnitzii and Foxp3 gene expression in GALT.
Conclusion: Our results suggest that the commensal microorganism B. fragilis may provide a protective role against the development of experimental colitis, which has to be taken into consideration for further clarification of the effective therapeutic strategy of inflammatory bowel diseases, irritable bowel syndrome and necrotising colitis.
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2. Ost K, Round J. Communication between the microbiota and mammalian immunity. Annu Rev Microbiol 2018; 72: 399-422.
3. Topol I, Kamyshny A. Study of expression of TLR2, TLR4 and transckription factor NF-kB structures of galt of rats in the conditions of the chronic social stress and modulation of structure of intestinal microflora. Georgian Med News 2013; (225): 115-122.
4. Topol IA, Kamyshny AM, Abramov AV, Kolesnik YM. Expression of XBP1 in lymphocytes of the small intestine in rats under chronic social stress and modulation of intestinal microflora composition. Fiziol Zh 2014; 60: 38-44.
5. Parker A, Lawson MAE, Vaux L, Pin C. Host-microbe interaction in the gastrointestinal tract. Environ Microbiol 2018; 20: 2337-2353.
6. McGuire VA, Arthur JS. Subverting Toll-like receptor signaling by bacterial pathogens. Front Immunol 2015; 6: 607.
7. Lee GR. The balance of Th17 versus treg cellsin autoimmunity. Int J Mol Sci 2018; 19: E730.
8. Krynytska I, Marushchak М, Mikolenko A, Bob A, Smachylo I, Radetska L. Differential diagnosis of hepatopulmonary syndrome (HPS): portopulmonary hypertension (PPH) and hereditary hemorrhagic telangiectasia (HHT). Bosn J Basic Med Sci 2017; 17:276-285.
9. Littman DR, Rudensky AY. Th17 and regulatory T cells in mediating and restraining inflammation. Cell 2010; 140: 845-858.
10. Ohnmacht C, Park JH, Cording S, Wing JB, Atarashi K, Obata Y, et al. Mucosal immunology. The microbiota regulates type 2 immunity through RORγt+ T cells. Science 2015; 349: 989-993.
11. Panda SK, Colonna M. Innate lymphoid cells in mucosal immunity. Front Immunol 2019; 10: 861.
12. Pantazi E, Powell N. Group 3 ILCs: Peacekeepers or Troublemakers? What's Your Gut Telling You?! Front. Immunol 2019;10:676.
13. Wang S, Xia P, Chen Y, Qu Y, Xiong Z, Ye B, et al. Regulatory innate lymphoid cells control innate intestinal inflammation. Cell 2017;171:201-216.
14. Sorini C, Cardoso RF, Gagliani N, Villablanca EJ. Commensal bacteria-specific CD4+ T cell responses in health and disease. Front Immunol 2018; 9: 2667.
15. Yang Y, Torchinsky M, Gobert M, Xiong H, Xu M. Focused specificity of intestinal TH17 cells towards commensal bacterial antigens. Nature 2014; 510: 152-156.
16. Lopetuso L, Scaldaferri F, Petito V, Gasbarrini A. Commensal Clostridia: leading players in the maintenance of gut homeostasis. Gut Pathog 2013; 5: 23.
17. Atarashi K, Tanoue T, Shima T, Imaoka A, Kuwahara T, Momose Y, et al. Induction of colonic regulatory T cells by indigenous Clostridium species. Science 2011; 331: 337-341.
18. Atarashi K, Tanoue T, Oshima K, Suda W, Nagano Y, Nishikawa H, et al. Treg induction by a rationally selected mixture of Clostridia strains from the human microbiota. Nature 2013; 500: 232-236.
19. Breyner NM, Michon C, de Sousa CS, Vilas Boas PB, Chain F, Azevedo VA, et al. Microbial anti-inflammatory molecule (MAM) from Faecali bacterium prausnitzii shows a protective effect on DNBS and DSS-Induced colitis model in mice through inhibition of NF-κB pathway. Front Microbiol 2017; 8: 114.
20. Li Z, Deng H, Zhou Y, Tan Y, Wang X, Han Y, et al. Bioluminescence imaging to track bacteroides fragilis inhibition of vibrio parahaemolyticus infection in mice. Front Cell Infect Microbiol 2017; 7: 170.
21. Smith PM, Howitt MR, Panikov N. The microbial metabolites, short-chain fatty acids, regulate colonic Treg cell homeostasis. Science 2013; 341: 569-573.
22. 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. Pathol 2017; 14: 12-19.
23. Bukina YuV, Varynskyi BO, Voitovich AV, Koval GD, Kaplaushenko AG, Kamyshnyi OM. The definition of neutrophil extracellular traps and the concentration of short-chain fatty acids in salmonella-induced inflammation of the intestine against the background of vancomycin and bacteroides fragilis. Pathol 2018; 15: 10-17.
24. Plovier H, Everard A, Druart C, Depommier C, Van Hul M, Geurts L, et al. A purified membrane protein from Akkermansia muciniphila or the pasteurized bacterium improves metabolism in obese and diabetic mice. Nat Med 2017; 23: 107-113.
25. Rozemond H. Laboratory animal protection: the European Convention and the Dutch Act. Vet Q 1986; 8: 346-349.
26. Korpela K, Flint HJ, Johnstone AM, Lappi J, Poutanen K, Dewulf E. Gut microbiota signatures predict host and microbiota responses to dietary interventions in obese individuals. PLoS One 2014; 9(6):e90702.
27. Zherebiatiev AS, Kamyshnyi AM. Regional peculiarities of the distribution of innate and adaptive immune cells in different segments of the intestine as factor determining the localization of the pathological process. Eksp Klin Gastroenterol 2015; (2): 46-51.
28. Zherebiatiev AS, Kamyshnyi AM. Transcriptional regulators of T-lymphocyte differentiation and pattern recognition receptors expression by lymphocytes in the intestine in experimental oxazolone-induced colitis in rats and after administration of inhibitor of 3-hydroxy-3- methylglutaryl coenzyme reductase and antagonist of receptors of interleukin-1. Immunology 2015; 36: 139-144.
29. Sano T, Huang W, Hall JA, Yang Y, Chen A. An IL-23R/IL-22 circuit regulates epithelial serum amyloid A to promote local effector Th17 responses. Cell 2015; 163: 381-393.
30. Domingues RG, Hepworth MR. Immunoregulatory sensory circuits in group 3 innate lymphoid cell (ILC3) function and tissue homeostasis. Front Immunol 2020;11:116.
31. Qiu J, Guo X, Chen ZM, He L, Sonnenberg GF, Artis D, et al. Group 3 innate lymphoid cells inhibit T-cell-mediated intestinal inflammation through aryl hydrocarbon receptor signaling and regulation of microflora. Immunity 2013; 39:386-399.
32. Sonnenberg GF, Monticelli LA, Alenghat T, Fung TC, Hutnick NA, Kunisawa J, et al. Innate lymphoid cells promote anatomical containment of lymphoid-resident commensal bacteria. Science 2012;336:1321-1325.
33. Zheng Y, Valdez PA, Danilenko DM, Hu Y, Sa SM, Gong Q, et al. Interleukin-22 mediates early host defense against attaching and effacing bacterial pathogens. Nat Med 2008; 14:282-289.
34. Ivanov II, Atarashi K, Manel N, Brodie EL, Shima T, Karaoz U, et al. Induction of intestinal Th17 cells by segmented filamentous bacteria. Cell 2009; 139: 485-498.
35. Goto Y, Umesaki Y, Benno Y, Kiyono H. Epithelial glycosylation in gut homeostasis and inflammation. Nat Immunol 2016; 17: 1244-1251.
36. Round JL, Mazmanian SK. Inducible Foxp3+ regulatory T-cell development by a commensal bacterium of the intestinal microbiota. Proc Natl Acad Sci USA 2010; 107: 12204-12209.
37. Arpaia N, Campbell C, Fan X. Metabolites produced by commensal bacteria promote peripheral regulatory T-cell generation. Nature 2013; 504: 451-455.
38. Lee YK, Mehrabian P, Boyajian S, Wu WL, Selicha J, Vonderfecht S, et al. The protective role of bacteroides fragilis in a murine model of colitis-associated colorectal cancer. mSphere 2018; 3(6): e00587-18.
39. Furusawa Y, Obata Y, Fukuda S. Commensal microbe-derived butyrate induces the differentiation of colonic regulatory T cells. Nature 2013; 504: 446-450.
40. Zamani S, Taslimi R, Sarabi A, Jasemi S, Sechi LA, Feizabadi MM. Enterotoxigenic Bacteroides fragilis: A possible etiological candidate for bacterially-induced colorectal precancerous and cancerous lesions. Front Cell Infect Microbiol 2020; 9: 449.
41. Zamani S, Shariati SH, Zali MR, Aghdaei HA, Asiabar AS, Bokaie S, et al. Detection of enterotoxigenic Bacteroides fragilis in patients with ulcerative colitis. Gut Pathog 2017; 9: 53.
42. Sears CL, Geis AL, Housseau F. Bacteroides fragilis subverts mucosal biology: from symbiont to colon carcinogenesis. J Clin Invest 2014; 124: 4166-4172.
43. Nutsch KM, Hsieh CS. T cell tolerance and immunity to commensal bacteria. Curr Opin Immunol 2012; 24: 385-391.
44. Hooper LV, Littman DR, Macpherson AJ. Interactions between the microbiota and the immune system. Science 2012; 336: 1268-1273.
45. Agbor TA, McCormick BA. Salmonella effectors: important players modulating host cell function during infection. Cell Microbiol 2011; 13: 1858-1869.
46. Behnsen J, Perez-Lopez A, Nuccio SP, Raffatellu M. Exploiting host immunity: the Salmonella paradigm. Trends Immunol 2015; 36: 112-120.
47. Shin NR, Lee JC, Lee HY, Kim MS, Whon TW, Lee MS. An increase in the Akkermansia spp. population induced by metformin treatment improves glucose homeostasis in diet-induced obese mice. Gut 2014; 63: 727-735.
48. Feuerer M, Hill JA, Kretschmer K, von Boehmer H, Mathis D, Benoist C. Genomic definition of multiple ex vivo regulatory T cell subphenotypes. Proc Natl Acad Sci U S A 2010; 107: 5919-5924.
49. Geuking MB, Cahenzli J, Lawson MA, Ng DC, Slack E, Hapfelmeier S, et al. Intestinal bacterial colonization induces mutualistic regulatory T cell responses. Immunity 2011; 34: 794-806.
50. Tan TG, Sefik E, Geva-Zatorsky N, Kua L, Naskar D, Teng F, et al. Identifying species of symbiont bacteria from the human gut that, alone, can induce intestinal Th17 cells in mice. Proc Natl Acad Sci U S A 2016; 113: E8141-E8150.
51. Geva-Zatorsky N, Sefik E, Kua L, Pasman L, Tan TG, Ortiz-Lopez A, et al. Mining the human gut microbiota for immunomodulatory organisms. Cell 2017; 168: 928-943.e11.
2. Ost K, Round J. Communication between the microbiota and mammalian immunity. Annu Rev Microbiol 2018; 72: 399-422.
3. Topol I, Kamyshny A. Study of expression of TLR2, TLR4 and transckription factor NF-kB structures of galt of rats in the conditions of the chronic social stress and modulation of structure of intestinal microflora. Georgian Med News 2013; (225): 115-122.
4. Topol IA, Kamyshny AM, Abramov AV, Kolesnik YM. Expression of XBP1 in lymphocytes of the small intestine in rats under chronic social stress and modulation of intestinal microflora composition. Fiziol Zh 2014; 60: 38-44.
5. Parker A, Lawson MAE, Vaux L, Pin C. Host-microbe interaction in the gastrointestinal tract. Environ Microbiol 2018; 20: 2337-2353.
6. McGuire VA, Arthur JS. Subverting Toll-like receptor signaling by bacterial pathogens. Front Immunol 2015; 6: 607.
7. Lee GR. The balance of Th17 versus treg cellsin autoimmunity. Int J Mol Sci 2018; 19: E730.
8. Krynytska I, Marushchak М, Mikolenko A, Bob A, Smachylo I, Radetska L. Differential diagnosis of hepatopulmonary syndrome (HPS): portopulmonary hypertension (PPH) and hereditary hemorrhagic telangiectasia (HHT). Bosn J Basic Med Sci 2017; 17:276-285.
9. Littman DR, Rudensky AY. Th17 and regulatory T cells in mediating and restraining inflammation. Cell 2010; 140: 845-858.
10. Ohnmacht C, Park JH, Cording S, Wing JB, Atarashi K, Obata Y, et al. Mucosal immunology. The microbiota regulates type 2 immunity through RORγt+ T cells. Science 2015; 349: 989-993.
11. Panda SK, Colonna M. Innate lymphoid cells in mucosal immunity. Front Immunol 2019; 10: 861.
12. Pantazi E, Powell N. Group 3 ILCs: Peacekeepers or Troublemakers? What's Your Gut Telling You?! Front. Immunol 2019;10:676.
13. Wang S, Xia P, Chen Y, Qu Y, Xiong Z, Ye B, et al. Regulatory innate lymphoid cells control innate intestinal inflammation. Cell 2017;171:201-216.
14. Sorini C, Cardoso RF, Gagliani N, Villablanca EJ. Commensal bacteria-specific CD4+ T cell responses in health and disease. Front Immunol 2018; 9: 2667.
15. Yang Y, Torchinsky M, Gobert M, Xiong H, Xu M. Focused specificity of intestinal TH17 cells towards commensal bacterial antigens. Nature 2014; 510: 152-156.
16. Lopetuso L, Scaldaferri F, Petito V, Gasbarrini A. Commensal Clostridia: leading players in the maintenance of gut homeostasis. Gut Pathog 2013; 5: 23.
17. Atarashi K, Tanoue T, Shima T, Imaoka A, Kuwahara T, Momose Y, et al. Induction of colonic regulatory T cells by indigenous Clostridium species. Science 2011; 331: 337-341.
18. Atarashi K, Tanoue T, Oshima K, Suda W, Nagano Y, Nishikawa H, et al. Treg induction by a rationally selected mixture of Clostridia strains from the human microbiota. Nature 2013; 500: 232-236.
19. Breyner NM, Michon C, de Sousa CS, Vilas Boas PB, Chain F, Azevedo VA, et al. Microbial anti-inflammatory molecule (MAM) from Faecali bacterium prausnitzii shows a protective effect on DNBS and DSS-Induced colitis model in mice through inhibition of NF-κB pathway. Front Microbiol 2017; 8: 114.
20. Li Z, Deng H, Zhou Y, Tan Y, Wang X, Han Y, et al. Bioluminescence imaging to track bacteroides fragilis inhibition of vibrio parahaemolyticus infection in mice. Front Cell Infect Microbiol 2017; 7: 170.
21. Smith PM, Howitt MR, Panikov N. The microbial metabolites, short-chain fatty acids, regulate colonic Treg cell homeostasis. Science 2013; 341: 569-573.
22. 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. Pathol 2017; 14: 12-19.
23. Bukina YuV, Varynskyi BO, Voitovich AV, Koval GD, Kaplaushenko AG, Kamyshnyi OM. The definition of neutrophil extracellular traps and the concentration of short-chain fatty acids in salmonella-induced inflammation of the intestine against the background of vancomycin and bacteroides fragilis. Pathol 2018; 15: 10-17.
24. Plovier H, Everard A, Druart C, Depommier C, Van Hul M, Geurts L, et al. A purified membrane protein from Akkermansia muciniphila or the pasteurized bacterium improves metabolism in obese and diabetic mice. Nat Med 2017; 23: 107-113.
25. Rozemond H. Laboratory animal protection: the European Convention and the Dutch Act. Vet Q 1986; 8: 346-349.
26. Korpela K, Flint HJ, Johnstone AM, Lappi J, Poutanen K, Dewulf E. Gut microbiota signatures predict host and microbiota responses to dietary interventions in obese individuals. PLoS One 2014; 9(6):e90702.
27. Zherebiatiev AS, Kamyshnyi AM. Regional peculiarities of the distribution of innate and adaptive immune cells in different segments of the intestine as factor determining the localization of the pathological process. Eksp Klin Gastroenterol 2015; (2): 46-51.
28. Zherebiatiev AS, Kamyshnyi AM. Transcriptional regulators of T-lymphocyte differentiation and pattern recognition receptors expression by lymphocytes in the intestine in experimental oxazolone-induced colitis in rats and after administration of inhibitor of 3-hydroxy-3- methylglutaryl coenzyme reductase and antagonist of receptors of interleukin-1. Immunology 2015; 36: 139-144.
29. Sano T, Huang W, Hall JA, Yang Y, Chen A. An IL-23R/IL-22 circuit regulates epithelial serum amyloid A to promote local effector Th17 responses. Cell 2015; 163: 381-393.
30. Domingues RG, Hepworth MR. Immunoregulatory sensory circuits in group 3 innate lymphoid cell (ILC3) function and tissue homeostasis. Front Immunol 2020;11:116.
31. Qiu J, Guo X, Chen ZM, He L, Sonnenberg GF, Artis D, et al. Group 3 innate lymphoid cells inhibit T-cell-mediated intestinal inflammation through aryl hydrocarbon receptor signaling and regulation of microflora. Immunity 2013; 39:386-399.
32. Sonnenberg GF, Monticelli LA, Alenghat T, Fung TC, Hutnick NA, Kunisawa J, et al. Innate lymphoid cells promote anatomical containment of lymphoid-resident commensal bacteria. Science 2012;336:1321-1325.
33. Zheng Y, Valdez PA, Danilenko DM, Hu Y, Sa SM, Gong Q, et al. Interleukin-22 mediates early host defense against attaching and effacing bacterial pathogens. Nat Med 2008; 14:282-289.
34. Ivanov II, Atarashi K, Manel N, Brodie EL, Shima T, Karaoz U, et al. Induction of intestinal Th17 cells by segmented filamentous bacteria. Cell 2009; 139: 485-498.
35. Goto Y, Umesaki Y, Benno Y, Kiyono H. Epithelial glycosylation in gut homeostasis and inflammation. Nat Immunol 2016; 17: 1244-1251.
36. Round JL, Mazmanian SK. Inducible Foxp3+ regulatory T-cell development by a commensal bacterium of the intestinal microbiota. Proc Natl Acad Sci USA 2010; 107: 12204-12209.
37. Arpaia N, Campbell C, Fan X. Metabolites produced by commensal bacteria promote peripheral regulatory T-cell generation. Nature 2013; 504: 451-455.
38. Lee YK, Mehrabian P, Boyajian S, Wu WL, Selicha J, Vonderfecht S, et al. The protective role of bacteroides fragilis in a murine model of colitis-associated colorectal cancer. mSphere 2018; 3(6): e00587-18.
39. Furusawa Y, Obata Y, Fukuda S. Commensal microbe-derived butyrate induces the differentiation of colonic regulatory T cells. Nature 2013; 504: 446-450.
40. Zamani S, Taslimi R, Sarabi A, Jasemi S, Sechi LA, Feizabadi MM. Enterotoxigenic Bacteroides fragilis: A possible etiological candidate for bacterially-induced colorectal precancerous and cancerous lesions. Front Cell Infect Microbiol 2020; 9: 449.
41. Zamani S, Shariati SH, Zali MR, Aghdaei HA, Asiabar AS, Bokaie S, et al. Detection of enterotoxigenic Bacteroides fragilis in patients with ulcerative colitis. Gut Pathog 2017; 9: 53.
42. Sears CL, Geis AL, Housseau F. Bacteroides fragilis subverts mucosal biology: from symbiont to colon carcinogenesis. J Clin Invest 2014; 124: 4166-4172.
43. Nutsch KM, Hsieh CS. T cell tolerance and immunity to commensal bacteria. Curr Opin Immunol 2012; 24: 385-391.
44. Hooper LV, Littman DR, Macpherson AJ. Interactions between the microbiota and the immune system. Science 2012; 336: 1268-1273.
45. Agbor TA, McCormick BA. Salmonella effectors: important players modulating host cell function during infection. Cell Microbiol 2011; 13: 1858-1869.
46. Behnsen J, Perez-Lopez A, Nuccio SP, Raffatellu M. Exploiting host immunity: the Salmonella paradigm. Trends Immunol 2015; 36: 112-120.
47. Shin NR, Lee JC, Lee HY, Kim MS, Whon TW, Lee MS. An increase in the Akkermansia spp. population induced by metformin treatment improves glucose homeostasis in diet-induced obese mice. Gut 2014; 63: 727-735.
48. Feuerer M, Hill JA, Kretschmer K, von Boehmer H, Mathis D, Benoist C. Genomic definition of multiple ex vivo regulatory T cell subphenotypes. Proc Natl Acad Sci U S A 2010; 107: 5919-5924.
49. Geuking MB, Cahenzli J, Lawson MA, Ng DC, Slack E, Hapfelmeier S, et al. Intestinal bacterial colonization induces mutualistic regulatory T cell responses. Immunity 2011; 34: 794-806.
50. Tan TG, Sefik E, Geva-Zatorsky N, Kua L, Naskar D, Teng F, et al. Identifying species of symbiont bacteria from the human gut that, alone, can induce intestinal Th17 cells in mice. Proc Natl Acad Sci U S A 2016; 113: E8141-E8150.
51. Geva-Zatorsky N, Sefik E, Kua L, Pasman L, Tan TG, Ortiz-Lopez A, et al. Mining the human gut microbiota for immunomodulatory organisms. Cell 2017; 168: 928-943.e11.
| Files | ||
| Issue | Vol 12 No 3 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v12i3.3241 | |
| Keywords | ||
| Salmonella; Vancomycin; Bacteroids; Immunoregulation; Genes expression | ||
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How to Cite
1.
Bukina Y, Thyhonovska M, Koval M, Marushchak M, Krynytska I, Kamyshnyi A. The effect of immunoregulatory bacteria on the transcriptional activity of Foxp3 and RORyt genes in the gut-associated lymphoid tissue with Salmonella-induced inflammation in the presence of vancomycin and Ваcteroides fragilis. Iran J Microbiol. 2020;12(3):231-241.
