title: TLR3-/4-Priming Differentially Promotes Ca(2+) Signaling and Cytokine Expression and Ca(2+)-Dependently Augments Cytokine Release in hMSCs.
abstract: In human mesenchymal stem cells (hMSCs), toll-like receptor 3 (TLR3) and TLR4 act as key players in the tissue repair process by recognizing their ligands and stimulating downstream processes including cytokine release. The mechanisms of TLR3- and TLR4-mediated cytokine releases from hMSCs remain uncertain. Here, we show that exposure to the TLR3 agonist polyinosinic-polycytidylic acid (poly(I:C)) or incubation with the TLR4 agonist lipopolysaccharide (LPS) increased the mRNA expression levels of TLR3, TLR4 and cytokines in hMSCs. Poly(I:C) exposure rather than LPS incubation not only elevated inositol 1,4,5-triphosphate receptor (IP3R) expression and IP3R-mediated Ca(2+) release, but also promoted Orai and STIM expression as well as store-operated Ca(2+) entry into hMSCs. In addition, we also observed that 21 Ca(2+) signaling genes were significantly up-regulated in response to TLR3 priming of hMSCs by RNA sequencing analysis. Both poly(I:C) and LPS exposure enhanced cytokine release from hMSCs. The enhanced cytokine release vanished upon siRNA knockdown and chelation of intracellular Ca(2+). These data demonstrate that TLR3- and TLR4-priming differentially enhance Ca(2+) signaling and cytokine expression, and Ca(2+) -dependently potentiates cytokine release in hMSCs.
authors.affiliation: Department of Molecular and Life Sciences, Hanyang University, Ansan, Korea.
authors.forename: Kyoung Sun
authors.initials: KS
authors.lastname: Park
authors.affiliation: Department of Molecular and Life Sciences, Hanyang University, Ansan, Korea.
authors.forename: Sun Hwa
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Department of Molecular and Life Sciences, Hanyang University, Ansan, Korea.
authors.forename: Amitabh
authors.initials: A
authors.lastname: Das
authors.affiliation: The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital L1, SE-171 76 Stockholm, Sweden.
authors.forename: Shao-Nian
authors.initials: SN
authors.lastname: Yang
authors.affiliation: Department of Molecular and Life Sciences, Hanyang University, Ansan, Korea.
authors.forename: Kyoung Hwa
authors.initials: KH
authors.lastname: Jung
authors.affiliation: Department of Physiology, Sungkyunkwan University School of Medicine, Suwon, Korea.
authors.forename: Mi Kyung
authors.initials: MK
authors.lastname: Kim
authors.affiliation: The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital L1, SE-171 76 Stockholm, Sweden.
authors.forename: Per-Olof
authors.initials: PO
authors.lastname: Berggren
authors.affiliation: Department of Molecular and Life Sciences, Hanyang University, Ansan, Korea.
authors.forename: YoungSeek
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Department of Molecular and Life Sciences, Hanyang University, Ansan, Korea.
authors.forename: Jin Choul
authors.initials: JC
authors.lastname: Chai
authors.affiliation: Department of Physiology, Sungkyunkwan University School of Medicine, Suwon, Korea.
authors.forename: Hyun Jin
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Molecular and Life Sciences, Hanyang University, Ansan, Korea.
authors.forename: Young Gyu
authors.initials: YG
authors.lastname: Chai
mesh_terms: D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000235:genetics*; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D000071740:ORAI1 Protein / Q000235:genetics / Q000378:metabolism; D011070:Poly I-C / Q000494:pharmacology; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071737:Stromal Interaction Molecule 1 / Q000235:genetics / Q000378:metabolism; D051196:Toll-Like Receptor 3 / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D055815:Young Adult
chemical_list: D016207:Cytokines; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008070:Lipopolysaccharides; D009363:Neoplasm Proteins; D000071740:ORAI1 Protein; C510746:ORAI1 protein, human; C103530:STIM1 protein, human; D000071737:Stromal Interaction Molecule 1; D051196:Toll-Like Receptor 3; D051197:Toll-Like Receptor 4; D011070:Poly I-C; D002118:Calcium