Abstract
The use of in vitro systems that allow efficient selection of probiotic candidates with immunomodulatory properties could significantly minimize the use of experimental animals. In this work, we generated an in vitro immunoassay system based on porcine intestinal epithelial (PIE) cells and dextran sodium sulfate (DSS) administration that could be useful for the selection and characterization of potential probiotic strains to be used in inflammatory bowel disease (IBD) patients. Our strategy was based on two fundamental pillars: on the one hand, the capacity of PIE cells to create a monolayer by attaching to neighboring cells and efficiently mount inflammatory responses and, on the other hand, the use of two probiotic bifidobacteria strains that have been characterized in terms of their immunomodulatory capacities, particularly in mouse IBD models and patients. Our results demonstrated that DSS administration can alter the epithelial barrier created in vitro by PIE cells and induce a potent inflammatory response, characterized by increases in the expression levels of several inflammatory factors including TNF-α, IL-1α, CCL4, CCL8, CCL11, CXCL5, CXCL9, CXCL10, SELL, SELE, EPCAM, VCAM, NCF2, and SAA2. In addition, we demonstrated that Bifidobacterium breve M-16V and B. longum BB536 are able to regulate the C-jun N-terminal kinase (JNK) intracellular signalling pathway, reducing the DSS-induced alterations of the in vitro epithelial barrier and differentially regulating the inflammatory response in a strain-dependent fashion. The good correlation between our in vitro findings in PIE cells and previous studies in animal models and IBD patients shows the potential value of our system to select new probiotic candidates in an efficient way.
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Funding
This study was supported by a Grant-in-Aid for Scientific Research (A) (19H00965), an Open Partnership Joint Projects of JSPS Bilateral Joint Research Projects from the Japan Society for the Promotion of Science (JSPS) and the Food Science Institute as well as the Foundation (Ryosyoku-Kenkyukai) to HK. This research was supported by grants from the project of NARO Bio-oriented Technology Research Advancement Institution (Research Program on the Development of Innovative Technology, No. 01002A) to HK. Md AI was supported by JSPS Postdoctoral Fellowship for Foreign Researchers, Program (No. 18F18081). This study was also supported by ANPCyT–FONCyT Grant PICT-2016-0410 to JV and by JSPS Core-to-Core Program, A. Advanced Research Networks entitled Establishment of International Agricultural Immunology Research-Core for a Quantum Improvement in Food Safety.
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Md Islam Aminul is a JSPS Postdoctoral Fellow.
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Sato, N., Yuzawa, M., Aminul, M.I. et al. Evaluation of Porcine Intestinal Epitheliocytes as an In vitro Immunoassay System for the Selection of Probiotic Bifidobacteria to Alleviate Inflammatory Bowel Disease. Probiotics & Antimicro. Prot. 13, 824–836 (2021). https://doi.org/10.1007/s12602-020-09694-z
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DOI: https://doi.org/10.1007/s12602-020-09694-z