Toll like receptor 3 & 4 responses of human turbinate derived mesenchymal stem cells: stimulation by double stranded RNA and lipopolysaccharide

PLoS One. 2014 Jul 8;9(7):e101558. doi: 10.1371/journal.pone.0101558. eCollection 2014.

Abstract

Background and objectives: 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.

Subjects and 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Surface / metabolism
  • Cell Differentiation
  • Cell Lineage
  • Cell Proliferation
  • Chemokines / biosynthesis
  • Coculture Techniques
  • Cytokines / biosynthesis
  • Humans
  • Immunophenotyping
  • Lipopolysaccharides / pharmacology
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / metabolism*
  • Phenotype
  • RNA, Double-Stranded / pharmacology
  • Toll-Like Receptor 3 / agonists
  • Toll-Like Receptor 3 / metabolism*
  • Toll-Like Receptor 4 / agonists
  • Toll-Like Receptor 4 / metabolism*
  • Turbinates / cytology*

Substances

  • Antigens, Surface
  • Chemokines
  • Cytokines
  • Lipopolysaccharides
  • RNA, Double-Stranded
  • Toll-Like Receptor 3
  • Toll-Like Receptor 4

Grants and funding

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by by the Ministry of Science, ICT and Future Planning (2014R1A2A2A01004325) and the Ministry of Education, Science and Technology (2010-0011249), Research Fund of Seoul St. Mary's Hospital, The Catholic University of Korea, Catholic Institute of Cell Therapy in 2013, the Otorhinolaryngology Alumni Fund of the Catholic University of Korea made in the program year of 2013, 2012 Research Fund for the Colleague of Catholic University Medical College, the financial support of the Catholic Medical Center Research Foundation made in the program year of 2013, and the financial support of the Catholic Medical Center Research Foundation made in the program year of 2014. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.