title: Toll like receptor 3 & 4 responses of human turbinate derived mesenchymal stem cells: stimulation by double stranded RNA and lipopolysaccharide.
abstract: OBJECTIVE
  Multipotent mesenchymal stromal cells (MSCs) represent a promising cell-based therapy for a number of inflammatory or autoimmune diseases. Herein, Toll like receptor (TLR) expression by MSCs and their immune regulatory roles are investigated. In this study, we investigated the influence of TLR on the immune response, proliferation, and differentiation potential of human turbinated MSC (hTMSC) cultures in vitro.
  
  
  METHODS
  After isolating hTMSCs from discarded inferior turbinate tissue, FACS analysis was used to assess the expression of TLRs such as TLR2, TLR3, TLR4, and TLR5 in hTMSCs and cell proliferation was assessed using a cell counting kit (CCK)-8. Cytokine and chemokine secretions were analyzed with multiplex immunoassays for IL-1α, IL-1β, IL-4, IL-6, IL-8, IL-10, IL-12, IP-10 (CXCL10), RANTES (CCL5), TNF-a, GM-CSF, and IFN-γ. The differentiation potential of hTMSCs was evaluated in the osteogenic, chondogenic, and adipogeinc media and analyzed by histology and gene expression related to differentiation.
  
  
  RESULTS
  FACS analysis revealed that TLR3 and TLR4 expression consisted of a relatively high percentage of the surface proteins expressed by hTMSCs. The proliferation of hTMSCs was influenced and significantly increased by the presence of TLR4 agonists. In particular, hTMSCs produced a set of cytokines and chemokines and the expression of IL-6, IL-8, IL-12, IP-10 (CXCL10), RANTES (CCL5), TNF-α, and GM-CSF were up-regulated in response to the TLR4 agonist LPS. The osteogenic and adipogeinc differentiation potential of hTMSCs was not affected by TLR agonists.
  
  
  CONCLUSIONS
  We conclude that TLR4 stimulation affects TLR expression, proliferation, and the immunomodulation potential of hTMSCs. Understanding the mechanism behind TLR's influence on hTMSCs and their immunomodulating properties would be useful for providing a novel target to exploit in the improvement of stem cell-based therapeutic strategies.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea.
authors.forename: Se Hwan
authors.initials: SH
authors.lastname: Hwang
authors.affiliation: Department of Pediatrics, Graduate School of Medicine, Gachon University, Incheon, Korea.
authors.forename: Hye Kyung
authors.initials: HK
authors.lastname: Cho
authors.affiliation: Institute of Clinical Medicine Research, College of Medicine, Catholic University of Korea, Seoul, Korea.
authors.forename: Sang Hi
authors.initials: SH
authors.lastname: Park
authors.affiliation: Institute of Clinical Medicine Research, College of Medicine, Catholic University of Korea, Seoul, Korea.
authors.forename: WeonSun
authors.initials: W
authors.lastname: Lee
authors.affiliation: Institute of Clinical Medicine Research, College of Medicine, Catholic University of Korea, Seoul, Korea.
authors.forename: Hee Jin
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea.
authors.forename: Dong Chang
authors.initials: DC
authors.lastname: Lee
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea.
authors.forename: Jeong Hoon
authors.initials: JH
authors.lastname: Oh
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea; Department of Biomedical Science, The Catholic University of Korea, College of Medicine, Seoul, Korea.
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Park
authors.affiliation: Catholic Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea; Department of Microbiology and Immunology, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Tai-Gyu
authors.initials: TG
authors.lastname: Kim
authors.affiliation: Catholic Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea.
authors.forename: Hyun-Jung
authors.initials: HJ
authors.lastname: Sohn
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea.
authors.forename: Jun Myung
authors.initials: JM
authors.lastname: Kang
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea; Department of Biomedical Science, The Catholic University of Korea, College of Medicine, Seoul, Korea.
authors.forename: Sung Won
authors.initials: SW
authors.lastname: Kim
mesh_terms: D000954:Antigens, Surface / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D018925:Chemokines / Q000096:biosynthesis; D018920:Coculture Techniques; D016207:Cytokines / Q000096:biosynthesis; D006801:Humans; D016130:Immunophenotyping; D008070:Lipopolysaccharides / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010641:Phenotype; D012330:RNA, Double-Stranded / Q000494:pharmacology; D051196:Toll-Like Receptor 3 / Q000819:agonists / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000819:agonists / Q000378:metabolism*; D014420:Turbinates / Q000166:cytology*
chemical_list: D000954:Antigens, Surface; D018925:Chemokines; D016207:Cytokines; D008070:Lipopolysaccharides; D012330:RNA, Double-Stranded; D051196:Toll-Like Receptor 3; D051197:Toll-Like Receptor 4