Immunomodulatory Effects of Adipose-derived Mesenchymal Stem Cells on Epithelial Cells Function in Response to Vibrio cholera in a Co-culture Model
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Abstract
Inflammation-induced by the interaction of the Vibrio cholerae with the epithelial cells is considered as a main cause of bacteria spreading through the gastrointestinal tract and its consequences. Because of the immunomodulatory and antibacterial properties of adipose-derived mesenchymal stem cells (AD-MSCs), this study aimed to investigate the effect of AD-MSCs on the interaction of the bacterial-epithelial cell.
Caco-2 differentiated to intestinal epithelial cells co-cultured with AD-MSCs in a 1:1 ratio of the surface area of six-well plates, for 48 hours. After exposure to Vibrio cholerae, bacterial attachment and internalization were evaluated. Secretions of interleukin (IL) -6, prostaglandin E2 (PGE2), and nitric oxide (NO) were also measured using ELISA, and Griess assay, respectively. In addition, the expression of chloratoxin (Ctx-β) and inflammatory cytokines such as TNF-α, IL-1β, and IL-8 were evaluated by real-time polymerase chain reaction (RT-PCR). The rate of apoptosis was also evaluated by Annexin V-PI flow cytometry.
Bacterial attachment and Ctx-β expression were significantly reduced in the co-culture group compared to the Vibrio cholerae-exposed Caco-2. IL-6 and PGE2 secretion increased in the co-culture group. NO, was also slightly reduced in exposure to Vibrio cholerae. An elevated level of bacterial internalization was observed in the co-culture group compared to the Caco-2 cells leading to an increase in the expression of pro-inflammatory cytokines. The rate of apoptosis was also increased significantly.
Cell-to-cell contact of AD-MSCs and Caco-2 promoted inflammatory responses and disruption of the epithelium barrier by enhancing bacterial invasion. This may be due to the high expression of surface matrix metalloproteinases on MSCs.
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2. Bishop AL, Patimalla B, Camilli A. Vibrio cholerae-induced inflammation in the neonatal mouse cholera model. Infect Immun. 2014;82(6):2434-47.
3. Molazade A, Soudi S, Bakhshi B. The Dual Role of Inflammatory Responses following Vibrio cholerae Infection in Cholerae pathogenesis. J Fasa Univ Med Sci. 2017;7(3):297-306.
4. Saeedi P, Halabian R, Fooladi AAI. Antimicrobial effects of mesenchymal stem cells primed by modified LPS on bacterial clearance in sepsis. J Cell Physiol. 2019;234(4):4970-86.
5. Domenis R, Cifù A, Quaglia S, Pistis C, Moretti M, Vicario A, et al. Pro inflammatory stimuli enhance the immunosuppressive functions of adipose mesenchymal stem cells-derived exosomes. Sci Rep. 2018;8(1):1-11.
6. Baharlou R, Rashidi N, Ahmadi-Vasmehjani A, Khoubyari M, Sheikh M, Erfanian S. Immunomodulatory effects of human adipose tissue-derived mesenchymal stem cells on T cell subsets in patients with rheumatoid arthritis. Iran J Allergy Asthma Immunol. 2019;18(1):114-9.
7. Sicco CL, Reverberi D, Balbi C, Ulivi V, Principi E, Pascucci L, et al. Mesenchymal stem cell‐derived extracellular vesicles as mediators of anti‐inflammatory effects: Endorsement of macrophage polarization. Stem Cells Transl Med. 2017;6(3):1018-28.
8. Nejad-Moghaddam A, Panahi Y, Alitappeh MA, Borna H, Shokrgozar MA, Ghanei M. Therapeutic potential of mesenchymal stem cells for the treatment of airway remodeling in pulmonary diseases. Iran J Allergy Asthma Immunol. 2015;14(6):552-68.
9. Dameshghi S, Zavaran-Hosseini A, Soudi S, Shirazi FJ, Nojehdehi S, Hashemi SM. Mesenchymal stem cells alter macrophage immune responses to Leishmania major infection in both susceptible and resistance mice. Immunol Lett. 2016;170:15-26.
10. Alamdary SZ, Bakhshi B, Soudi S. The anti-apoptotic and anti-inflammatory effect of Lactobacillus acidophilus on Shigella sonnei and Vibrio cholerae interaction with intestinal epithelial cells: A comparison between invasive and non-invasive bacteria. Plos one. 2018;13(6):e0196941.
11. Alamdary SZ, Bakhshi B. Lactobacillus acidophilus attenuates toxin production by Vibrio cholerae and shigella dysenteriae following intestinal epithelial cells infection. Microb Pathog. 2020;149(1):104543.
12. Philipp D, Suhr L, Wahlers T, Choi Y-H, Paunel-Görgülü A. Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization. Stem Cell Res Ther. 2018;9(1):1-17.
13. Piraghaj MG, Soudi S, Ghanbarian H, Bolandi Z, Namaki S, Hashemi SM. Effect of efferocytosis of apoptotic mesenchymal stem cells (MSCs) on C57BL/6 peritoneal macrophages function. Life Sci 2018;212(1):203-12.
14. Zanganeh E, Soudi S, Hosseini AZ, Khosrojerdi A. Repeated intravenous injection of adipose tissue derived mesenchymal stem cells enhances Th1 immune responses in Leishmania major-infected BALB/c mice. Immunol Lett. 2019;216:97-105.
15. Jiang F, Bi R, Deng L, Kang H, Gu B, Ma P. Virulence-associated genes and molecular characteristics of non-O1/non-O139 Vibrio cholerae isolated from hepatitis B cirrhosis patients in China. Int J Infect Dis 2018;74(1):117-22.
16. Banijamali RS, Soleimanjahi H, Soudi S, Karimi H. The Effect of Oncolytic Reovirus Infection on Nitric Oxide Secretion and Induction of Apoptosis in Adipose Tissue-Derived Mesenchymal Stem Cells. Iran J Med Microbiol. 2018;12(3):218-29.
17. Williams JM, Duckworth CA, Burkitt MD, Watson AJM, Campbell BJ, Pritchard DM. Epithelial cell shedding and barrier function: a matter of life and death at the small intestinal villus tip. Vet Pathol. 2015;52(3):445-55.
18. Saberpour M, Bakhshi B, Najar-Peerayeh S. Evaluation of the Antimicrobial and Antibiofilm Effect of Chitosan Nanoparticles as Carrier for Supernatant of Mesenchymal Stem Cells on Multidrug-Resistant Vibrio cholerae. Infect Drug Resist. 2020;13:2251.
19. Fiore A, Murray PJ. Tryptophan and indole metabolism in immune regulation. Curr Opin Immunol. 2021;70(1):7-14.
20. Mazini L, Rochette L, Admou B, Amal S, Malka G. Hopes and limits of adipose-derived stem cells (ADSCs) and mesenchymal stem cells (MSCs) in wound healing. Int J Mol Sci. 2020;21(4):1-19.
21. Saeedi P, Halabian R, Fooladi AAI. Mesenchymal stem cells preconditioned by staphylococcal enterotoxin B enhance survival and bacterial clearance in murine sepsis model. Cytotherapy. 2019;21(1):41-53.
22. Kuhn KA, Schulz HM, Regner EH, Severs EL, Hendrickson JD, Mehta G, et al. Bacteroidales recruit IL-6-producing intraepithelial lymphocytes in the colon to promote barrier integrity. Mucosal Immunol. 2018;11(2):357-68.
23. Ogata A, Kato Y, Higa S, Yoshizaki K. IL-6 inhibitor for the treatment of rheumatoid arthritis: a comprehensive review. Mod Rheumatol. 2019;29(2):258-67.
24. Bandyopadhaya A, Sarkar M, Chaudhuri K. Transcriptional upregulation of inflammatory cytokines in human intestinal epithelial cells following Vibrio cholerae infection. FEBS J. 2007;274(17):4631-42.
25. Tsuge K, Inazumi T, Shimamoto A, Sugimoto Y. Molecular mechanisms underlying prostaglandin E2-exacerbated inflammation and immune diseases. Int Immunol. 2019;31(9):597-606.
26. Speelman P, Rabbani GH, Bukhave K, Rask-Madsen J. Increased jejunal prostaglandin E2 concentrations in patients with acute cholera. Gut. 1985;26(2):188-93.
27. Hausmann M. How bacteria-induced apoptosis of intestinal epithelial cells contributes to mucosal inflammation. Int J Inflam. 2010;2010(1):1-9.
28. Johnson V, Webb T, Norman A, Coy J, Kurihara J, Regan D, et al. Activated mesenchymal stem cells interact with antibiotics and host innate immune responses to control chronic bacterial infections. Sci Rep. 2017;7(1):1-18.
29. Najar M, Krayem M, Merimi M, Burny A, Meuleman N, Bron D, et al. Insights into inflammatory priming of mesenchymal stromal cells: functional biological impacts. Inflamm Res. 2018;67(6):467-77.
30. Hyldig K, Riis S, Pennisi CP, Zachar V, Fink T. Implications of extracellular matrix production by adipose tissue-derived stem cells for development of wound healing therapies. Int J Mol Sci. 2017;18(6):1167.
31. Ramamurthy T, Nandy RK, Mukhopadhyay AK, Dutta S, Mutreja A, Okamoto K, et al. Virulence Regulation and Innate Host Response in the Pathogenicity of Vibrio cholerae. Front Cell Infect Microbiol. 2020;10(1):1-22.
32. Ghasemi M, Bakhshi B, Khashei R, Soudi S. Modulatory effect of Vibrio cholerae toxin co-regulated pilus on mucins, toll-like receptors and NOD genes expression in co-culture model of Caco-2 and peripheral blood mononuclear cells (PBMC). Microb Pathog. 2020;149(1):1-19.
33. Vancamelbeke M, Vermeire S. The intestinal barrier: a fundamental role in health and disease. Expert Rev Gastroenterol Hepatol. 2017;11(9):821-34.
34. Kiesslich R, Goetz M, Angus EM, Hu Q, Guan Y, Potten C, et al. Identification of epithelial gaps in human small and large intestine by confocal endomicroscopy. Gastroenterology. 2007;133(6):1769-78.
35. Ma TY, Iwamoto GK, Hoa NT, Akotia V, Pedram A, Boivin MA, et al. TNF-α-induced increase in intestinal epithelial tight junction permeability requires NF-κB activation. Am J Physiol Gastrointest Liver Physiol. 2004;286(3):G367-76.
36. Al-Sadi R, Guo S, Ye D, Dokladny K, Alhmoud T, Ereifej L, et al. Mechanism of IL-1β modulation of intestinal epithelial barrier involves p38 kinase and activating transcription factor-2 activation. J Immunol. 2013;190(12):6596-606.
2. Bishop AL, Patimalla B, Camilli A. Vibrio cholerae-induced inflammation in the neonatal mouse cholera model. Infect Immun. 2014;82(6):2434-47.
3. Molazade A, Soudi S, Bakhshi B. The Dual Role of Inflammatory Responses following Vibrio cholerae Infection in Cholerae pathogenesis. J Fasa Univ Med Sci. 2017;7(3):297-306.
4. Saeedi P, Halabian R, Fooladi AAI. Antimicrobial effects of mesenchymal stem cells primed by modified LPS on bacterial clearance in sepsis. J Cell Physiol. 2019;234(4):4970-86.
5. Domenis R, Cifù A, Quaglia S, Pistis C, Moretti M, Vicario A, et al. Pro inflammatory stimuli enhance the immunosuppressive functions of adipose mesenchymal stem cells-derived exosomes. Sci Rep. 2018;8(1):1-11.
6. Baharlou R, Rashidi N, Ahmadi-Vasmehjani A, Khoubyari M, Sheikh M, Erfanian S. Immunomodulatory effects of human adipose tissue-derived mesenchymal stem cells on T cell subsets in patients with rheumatoid arthritis. Iran J Allergy Asthma Immunol. 2019;18(1):114-9.
7. Sicco CL, Reverberi D, Balbi C, Ulivi V, Principi E, Pascucci L, et al. Mesenchymal stem cell‐derived extracellular vesicles as mediators of anti‐inflammatory effects: Endorsement of macrophage polarization. Stem Cells Transl Med. 2017;6(3):1018-28.
8. Nejad-Moghaddam A, Panahi Y, Alitappeh MA, Borna H, Shokrgozar MA, Ghanei M. Therapeutic potential of mesenchymal stem cells for the treatment of airway remodeling in pulmonary diseases. Iran J Allergy Asthma Immunol. 2015;14(6):552-68.
9. Dameshghi S, Zavaran-Hosseini A, Soudi S, Shirazi FJ, Nojehdehi S, Hashemi SM. Mesenchymal stem cells alter macrophage immune responses to Leishmania major infection in both susceptible and resistance mice. Immunol Lett. 2016;170:15-26.
10. Alamdary SZ, Bakhshi B, Soudi S. The anti-apoptotic and anti-inflammatory effect of Lactobacillus acidophilus on Shigella sonnei and Vibrio cholerae interaction with intestinal epithelial cells: A comparison between invasive and non-invasive bacteria. Plos one. 2018;13(6):e0196941.
11. Alamdary SZ, Bakhshi B. Lactobacillus acidophilus attenuates toxin production by Vibrio cholerae and shigella dysenteriae following intestinal epithelial cells infection. Microb Pathog. 2020;149(1):104543.
12. Philipp D, Suhr L, Wahlers T, Choi Y-H, Paunel-Görgülü A. Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization. Stem Cell Res Ther. 2018;9(1):1-17.
13. Piraghaj MG, Soudi S, Ghanbarian H, Bolandi Z, Namaki S, Hashemi SM. Effect of efferocytosis of apoptotic mesenchymal stem cells (MSCs) on C57BL/6 peritoneal macrophages function. Life Sci 2018;212(1):203-12.
14. Zanganeh E, Soudi S, Hosseini AZ, Khosrojerdi A. Repeated intravenous injection of adipose tissue derived mesenchymal stem cells enhances Th1 immune responses in Leishmania major-infected BALB/c mice. Immunol Lett. 2019;216:97-105.
15. Jiang F, Bi R, Deng L, Kang H, Gu B, Ma P. Virulence-associated genes and molecular characteristics of non-O1/non-O139 Vibrio cholerae isolated from hepatitis B cirrhosis patients in China. Int J Infect Dis 2018;74(1):117-22.
16. Banijamali RS, Soleimanjahi H, Soudi S, Karimi H. The Effect of Oncolytic Reovirus Infection on Nitric Oxide Secretion and Induction of Apoptosis in Adipose Tissue-Derived Mesenchymal Stem Cells. Iran J Med Microbiol. 2018;12(3):218-29.
17. Williams JM, Duckworth CA, Burkitt MD, Watson AJM, Campbell BJ, Pritchard DM. Epithelial cell shedding and barrier function: a matter of life and death at the small intestinal villus tip. Vet Pathol. 2015;52(3):445-55.
18. Saberpour M, Bakhshi B, Najar-Peerayeh S. Evaluation of the Antimicrobial and Antibiofilm Effect of Chitosan Nanoparticles as Carrier for Supernatant of Mesenchymal Stem Cells on Multidrug-Resistant Vibrio cholerae. Infect Drug Resist. 2020;13:2251.
19. Fiore A, Murray PJ. Tryptophan and indole metabolism in immune regulation. Curr Opin Immunol. 2021;70(1):7-14.
20. Mazini L, Rochette L, Admou B, Amal S, Malka G. Hopes and limits of adipose-derived stem cells (ADSCs) and mesenchymal stem cells (MSCs) in wound healing. Int J Mol Sci. 2020;21(4):1-19.
21. Saeedi P, Halabian R, Fooladi AAI. Mesenchymal stem cells preconditioned by staphylococcal enterotoxin B enhance survival and bacterial clearance in murine sepsis model. Cytotherapy. 2019;21(1):41-53.
22. Kuhn KA, Schulz HM, Regner EH, Severs EL, Hendrickson JD, Mehta G, et al. Bacteroidales recruit IL-6-producing intraepithelial lymphocytes in the colon to promote barrier integrity. Mucosal Immunol. 2018;11(2):357-68.
23. Ogata A, Kato Y, Higa S, Yoshizaki K. IL-6 inhibitor for the treatment of rheumatoid arthritis: a comprehensive review. Mod Rheumatol. 2019;29(2):258-67.
24. Bandyopadhaya A, Sarkar M, Chaudhuri K. Transcriptional upregulation of inflammatory cytokines in human intestinal epithelial cells following Vibrio cholerae infection. FEBS J. 2007;274(17):4631-42.
25. Tsuge K, Inazumi T, Shimamoto A, Sugimoto Y. Molecular mechanisms underlying prostaglandin E2-exacerbated inflammation and immune diseases. Int Immunol. 2019;31(9):597-606.
26. Speelman P, Rabbani GH, Bukhave K, Rask-Madsen J. Increased jejunal prostaglandin E2 concentrations in patients with acute cholera. Gut. 1985;26(2):188-93.
27. Hausmann M. How bacteria-induced apoptosis of intestinal epithelial cells contributes to mucosal inflammation. Int J Inflam. 2010;2010(1):1-9.
28. Johnson V, Webb T, Norman A, Coy J, Kurihara J, Regan D, et al. Activated mesenchymal stem cells interact with antibiotics and host innate immune responses to control chronic bacterial infections. Sci Rep. 2017;7(1):1-18.
29. Najar M, Krayem M, Merimi M, Burny A, Meuleman N, Bron D, et al. Insights into inflammatory priming of mesenchymal stromal cells: functional biological impacts. Inflamm Res. 2018;67(6):467-77.
30. Hyldig K, Riis S, Pennisi CP, Zachar V, Fink T. Implications of extracellular matrix production by adipose tissue-derived stem cells for development of wound healing therapies. Int J Mol Sci. 2017;18(6):1167.
31. Ramamurthy T, Nandy RK, Mukhopadhyay AK, Dutta S, Mutreja A, Okamoto K, et al. Virulence Regulation and Innate Host Response in the Pathogenicity of Vibrio cholerae. Front Cell Infect Microbiol. 2020;10(1):1-22.
32. Ghasemi M, Bakhshi B, Khashei R, Soudi S. Modulatory effect of Vibrio cholerae toxin co-regulated pilus on mucins, toll-like receptors and NOD genes expression in co-culture model of Caco-2 and peripheral blood mononuclear cells (PBMC). Microb Pathog. 2020;149(1):1-19.
33. Vancamelbeke M, Vermeire S. The intestinal barrier: a fundamental role in health and disease. Expert Rev Gastroenterol Hepatol. 2017;11(9):821-34.
34. Kiesslich R, Goetz M, Angus EM, Hu Q, Guan Y, Potten C, et al. Identification of epithelial gaps in human small and large intestine by confocal endomicroscopy. Gastroenterology. 2007;133(6):1769-78.
35. Ma TY, Iwamoto GK, Hoa NT, Akotia V, Pedram A, Boivin MA, et al. TNF-α-induced increase in intestinal epithelial tight junction permeability requires NF-κB activation. Am J Physiol Gastrointest Liver Physiol. 2004;286(3):G367-76.
36. Al-Sadi R, Guo S, Ye D, Dokladny K, Alhmoud T, Ereifej L, et al. Mechanism of IL-1β modulation of intestinal epithelial barrier involves p38 kinase and activating transcription factor-2 activation. J Immunol. 2013;190(12):6596-606.
| Files | ||
| Issue | Vol 20 No 5 (2021) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijaai.v20i5.7405 | |
| Keywords | ||
| Apoptosis Caco-2 cells InterleukinL-6 Mesenchymal stem cells Vibrio cholerae Virus internalization | ||
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How to Cite
1.
Moulazadeh A, Soudi S, Bakhshi B. Immunomodulatory Effects of Adipose-derived Mesenchymal Stem Cells on Epithelial Cells Function in Response to Vibrio cholera in a Co-culture Model. Iran J Allergy Asthma Immunol. 2021;20(5):550-562.
