title: The toll-like receptor family protein RP105/MD1 complex is involved in the immunoregulatory effect of exopolysaccharides from Lactobacillus plantarum N14.
abstract: The radioprotective 105 (RP105)/MD1 complex is a member of the Toll-like receptor (TLR) family. It was reported that RP105/MD1 cooperates with the lipopolysaccharide (LPS) receptor TLR4/MD2 complex and plays a crucial role in the response of immune cells to LPS. This work evaluated whether RP105, TLR4 or TLR2 were involved in the immunoregulatory capacities of Lactobacillus plantarum N14 (LP14) or its exopolysaccharides (EPS). EPS from LP14 were fractionated into neutral (NPS) and acidic (APS) EPS by anion exchange chromatography. Experiments with transfectant HEK(RP105/MD1) and HEK(TLR2) cells demonstrated that LP14 strongly activated NF-κB via RP105 and TLR2. When we studied the capacity of APS to activate NF-κB pathway in HEK(RP105/MD1) and HEK(TLR4) cells; we observed that APS strongly stimulated both transfectant cells. Our results also showed that LP14 and APS were able to decrease the production of pro-inflammatory cytokines (IL-6, IL-8 and MCP-1) in porcine intestinal epithelial (PIE) cells in response to enterotoxigenic Escherichia coli (ETEC) challenge. In order to confirm the role of TLR2, TLR4 and RP105 in the immunoregulatory effect of APS from LP14, we used small interfering RNA (siRNA) to knockdown these receptors in PIE cells. The capacity of LP14 and APS to modulate pro-inflammatory cytokine expression was significantly reduced in PIE(RP105-/-) cells. It was also shown that LP14 and APS were capable of upregulating negative regulators of the TLR signaling in PIE cells. This work describes for the first time that a Lactobacillus strain and its EPS reduce inflammation in intestinal epithelial cells in a RP105/MD1-dependend manner.
authors.affiliation: Food and Feed Immunology Group, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan. Electronic address: sunnytrump.60@gmail.com.
authors.forename: Yo
authors.initials: Y
authors.lastname: Murofushi
authors.affiliation: Food and Feed Immunology Group, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan; Laboratory of Immunobiotechnology, CERELA-CONICET, Tucuman, Argentina. Electronic address: jcvillena@cerela.org.ar.
authors.forename: Julio
authors.initials: J
authors.lastname: Villena
authors.affiliation: Food and Feed Immunology Group, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan. Electronic address: st205miffy@yahoo.co.jp.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Morie
authors.affiliation: Food and Feed Immunology Group, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan. Electronic address: kanmanibiotech2007@gmail.com.
authors.forename: Paulraj
authors.initials: P
authors.lastname: Kanmani
authors.affiliation: Functional Feed Research Team, National Institute of Livestock and Grassland Science, National Agricultural Research Organization, Nasushiobara, Tochigi 329-2793, Japan. Electronic address: tohno@affrc.go.jp.
authors.forename: Masanori
authors.initials: M
authors.lastname: Tohno
authors.affiliation: Laboratory of Animal Breeding and Genetics, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan. Electronic address: tshimazu@rs.tus.ac.jp.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Shimazu
authors.affiliation: Cell Biology Laboratory, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan. Electronic address: asosan@bios.tohoku.ac.jp.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Aso
authors.affiliation: Department of Food, Agriculture and Environment, Miyagi University, Sendai 982-0215, Japan. Electronic address: suda@myu.ac.jp.
authors.forename: Yoshihito
authors.initials: Y
authors.lastname: Suda
authors.affiliation: Research Center, Momoya Co., Ltd, Saitama 344-8522, Japan. Electronic address: hashiguchi@momoya.co.jp.
authors.forename: Kenji
authors.initials: K
authors.lastname: Hashiguchi
authors.affiliation: Food and Feed Immunology Group, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan. Electronic address: tsaito@bios.tohoku.ac.jp.
authors.forename: Tadao
authors.initials: T
authors.lastname: Saito
authors.affiliation: Food and Feed Immunology Group, Graduate School of Agricultural Science, Tohoku University, Aobaku, Sendai 981-8555, Japan. Electronic address: haruki@bios.tohoku.ac.jp.
authors.forename: Haruki
authors.initials: H
authors.lastname: Kitazawa
mesh_terms: D000143:Acids; D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D015703:Antigens, CD / Q000378:metabolism*; D005591:Chemical Fractionation; D020895:Enterocytes / Q000187:drug effects / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D056747:Immunomodulation / Q000187:drug effects*; D048191:Lactiplantibacillus plantarum / Q000737:chemistry*; D008198:Lymph Nodes / Q000187:drug effects / Q000378:metabolism; D010581:Peyer's Patches / Q000187:drug effects / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D034421:Sus scrofa; D051195:Toll-Like Receptor 2 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D000143:Acids; D019138:Antibodies, Blocking; D015703:Antigens, CD; D011135:Polysaccharides, Bacterial; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4
keywords: Exopolysaccharides; Lactobacillus plantarum N14; PIE cells; RP105/MD1; TLR2; TLR4