Inhibition of Lipopolysaccharide-Induced Interleukin 8 in Human Adenocarcinoma Cell Line HT-29 by Spore Probiotics: B. Coagulans and B. Subtilis (natto)

Overview

Journal Probiotics Antimicrob Proteins

Publisher Springer

Specialties Infectious Diseases
Microbiology
Nutritional Sciences
Pharmacology

Date 2016 Oct 28

PMID 27785697

Citations 5

Authors

Masoumeh Azimirad

Masoud Alebouyeh

Tahereh Naji

Affiliations

Soon will be listed here.

Abstract

Probiotics are used as a treatment for different intestinal disorders. They confer health benefits by different ways. This study was aimed to investigate immunomodulatory effect of Bacillus probiotic spores on the production of lipopolysaccharide (LPS)-induced interleukin 8 (IL-8) in HT-29 intestinal epithelial cells. Differentiated intestinal epithelial cell line was used as a model for the study of colonization of purified spores (Bacillus subtilis (natto) and B. coagulans) and their anti-inflammatory effects. MTT assay and trypan blue staining were used for the detection of optimal concentration of the purified spores and LPS. Pre-treatment assay was done by treatment of the cells with the purified spores for 2 h, followed by challenges with LPS for 3 and 18 h. Post-treatment assay was done by initial treatment of the cells with LPS for 18 h, followed by the spores for 3 and 6 h. Levels of IL-8 secretion and its mRNA expression were measured by ELISA and relative Q real-time PCR. Our results showed similar rates of adherence to intestinal epithelial cells by the spore probiotics, while displaying no cytotoxic effect. In the pre-treatment assay, a significant decrease in IL-8, at both protein and mRNA levels, was measured for B. coagulans spores after the addition of LPS, which was higher than those observed for Bacillus subtilis (natto) spores. In the post-treatment assay, while Bacillus subtilis (but not B. coagulans) diminished the LPS-stimulated IL-8 levels after 3 h of incubation, the inhibitory effect was not constant. In conclusion, ability of Bacillus spore probiotics for adherence to intestinal epithelial cell and their anti-inflammatory effects, through interference with LPS/IL-8 signaling, was shown in this study. Further studies are needed to characterize responsible bacterial compounds associated with these effects.

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References

Fedorak R, Madsen K . Probiotics and the management of inflammatory bowel disease. Inflamm Bowel Dis. 2004; 10(3):286-99. DOI: 10.1097/00054725-200405000-00018. View

Sanchez B, Arias S, Chaignepain S, Denayrolles M, Schmitter J, Bressollier P . Identification of surface proteins involved in the adhesion of a probiotic Bacillus cereus strain to mucin and fibronectin. Microbiology (Reading). 2009; 155(Pt 5):1708-1716. DOI: 10.1099/mic.0.025288-0. View

Duc L, Hong H, Fairweather N, Ricca E, Cutting S . Bacterial spores as vaccine vehicles. Infect Immun. 2003; 71(5):2810-8. PMC: 153275. DOI: 10.1128/IAI.71.5.2810-2818.2003. View

Zughaier S, Zimmer S, Datta A, Carlson R, Stephens D . Differential induction of the toll-like receptor 4-MyD88-dependent and -independent signaling pathways by endotoxins. Infect Immun. 2005; 73(5):2940-50. PMC: 1087371. DOI: 10.1128/IAI.73.5.2940-2950.2005. View

Hardy H, Harris J, Lyon E, Beal J, Foey A . Probiotics, prebiotics and immunomodulation of gut mucosal defences: homeostasis and immunopathology. Nutrients. 2013; 5(6):1869-912. PMC: 3725482. DOI: 10.3390/nu5061869. View

Moue M, Tohno M, Shimazu T, Kido T, Aso H, Saito T . Toll-like receptor 4 and cytokine expression involved in functional immune response in an originally established porcine intestinal epitheliocyte cell line. Biochim Biophys Acta. 2007; 1780(2):134-44. DOI: 10.1016/j.bbagen.2007.11.006. View

Medvedev A, Kopydlowski K, Vogel S . Inhibition of lipopolysaccharide-induced signal transduction in endotoxin-tolerized mouse macrophages: dysregulation of cytokine, chemokine, and toll-like receptor 2 and 4 gene expression. J Immunol. 2000; 164(11):5564-74. DOI: 10.4049/jimmunol.164.11.5564. View

Benson K, Redman K, Carter S, Keller D, Farmer S, Endres J . Probiotic metabolites from Bacillus coagulans GanedenBC30™ support maturation of antigen-presenting cells in vitro. World J Gastroenterol. 2012; 18(16):1875-83. PMC: 3337562. DOI: 10.3748/wjg.v18.i16.1875. View

Hosoi T, Hirose R, Saegusa S, Ametani A, Kiuchi K, Kaminogawa S . Cytokine responses of human intestinal epithelial-like Caco-2 cells to the nonpathogenic bacterium Bacillus subtilis (natto). Int J Food Microbiol. 2003; 82(3):255-64. DOI: 10.1016/s0168-1605(02)00311-2. View

10.

Blaut M, Clavel T . Metabolic diversity of the intestinal microbiota: implications for health and disease. J Nutr. 2007; 137(3 Suppl 2):751S-5S. DOI: 10.1093/jn/137.3.751S. View

11.

Finamore A, Roselli M, Imbinto A, Seeboth J, Oswald I, Mengheri E . Lactobacillus amylovorus inhibits the TLR4 inflammatory signaling triggered by enterotoxigenic Escherichia coli via modulation of the negative regulators and involvement of TLR2 in intestinal Caco-2 cells and pig explants. PLoS One. 2014; 9(4):e94891. PMC: 3986366. DOI: 10.1371/journal.pone.0094891. View

12.

La Ragione R, Casula G, Cutting S, Woodward M . Bacillus subtilis spores competitively exclude Escherichia coli O78:K80 in poultry. Vet Microbiol. 2001; 79(2):133-42. DOI: 10.1016/s0378-1135(00)00350-3. View

13.

Lenoir-Wijnkoop I, Sanders M, Cabana M, Caglar E, Corthier G, Rayes N . Probiotic and prebiotic influence beyond the intestinal tract. Nutr Rev. 2007; 65(11):469-89. DOI: 10.1111/j.1753-4887.2007.tb00272.x. View

14.

de Souza R, Batista M, Luiz W, Marques Cavalcante R, Amorim J, Bizerra R . Bacillus subtilis spores as vaccine adjuvants: further insights into the mechanisms of action. PLoS One. 2014; 9(1):e87454. PMC: 3903701. DOI: 10.1371/journal.pone.0087454. View

15.

Faille C, Jullien C, Fontaine F, Bellon-Fontaine M, Slomianny C, Benezech T . Adhesion of Bacillus spores and Escherichia coli cells to inert surfaces: role of surface hydrophobicity. Can J Microbiol. 2002; 48(8):728-38. DOI: 10.1139/w02-063. View

16.

Sabet M, Cottam H, Guiney D . Modulation of cytokine production and enhancement of cell viability by TLR7 and TLR9 ligands during anthrax infection of macrophages. FEMS Immunol Med Microbiol. 2006; 47(3):369-79. DOI: 10.1111/j.1574-695X.2006.00096.x. View

17.

Tekin I, Oral H, Shiri-Sverdlov R . Mediators of gut mucosal immunity and inflammation. Mediators Inflamm. 2015; 2015:765303. PMC: 4417598. DOI: 10.1155/2015/765303. View

18.

Taverniti V, Guglielmetti S . The immunomodulatory properties of probiotic microorganisms beyond their viability (ghost probiotics: proposal of paraprobiotic concept). Genes Nutr. 2011; 6(3):261-74. PMC: 3145061. DOI: 10.1007/s12263-011-0218-x. View

19.

Joshi L, Phillips D, Williams C, Alyousef A, Baillie L . Contribution of spores to the ability of Clostridium difficile to adhere to surfaces. Appl Environ Microbiol. 2012; 78(21):7671-9. PMC: 3485709. DOI: 10.1128/AEM.01862-12. View

20.

Cario E, ROSENBERG I, Brandwein S, Beck P, Reinecker H, Podolsky D . Lipopolysaccharide activates distinct signaling pathways in intestinal epithelial cell lines expressing Toll-like receptors. J Immunol. 2000; 164(2):966-72. DOI: 10.4049/jimmunol.164.2.966. View