title: Adipose-Derived Stem Cell Modulation of T-Cell Regulation Correlates with Heme Oxgenase-1 Pathway Changes.
abstract: BACKGROUND
  The authors' previous proteome study revealed that haptoglobin was involved in adipose-derived stem cell modulation of allotransplant survival and T-cell regulation to induce immune tolerance. This study investigated whether adipose-derived stem cells could modulate T-cell regulation through haptoglobin and the downstream heme oxgenase-1 pathway in vitro.
  
  
  METHODS
  Splenocytes were isolated from Lewis rat spleens and then CD3 T cells were purified using anti-CD3 beads. Adipose-derived stem cells were harvested from Lewis rats and co-cultured with the T cells. After Transwell co-culture at different periods, the authors analyzed cell proliferation with a bromodeoxyuridine assay. Cell extractions and culture supernatants were collected for further analysis. Heme oxgenase-1 and related protein expression levels from the adipose-derived stem cells and T cells were detected using Western blotting. The related cytokine expression levels were analyzed with enzyme-linked immunosorbent assay kits. Flow cytometry was used to detect the regulatory T-cell proportion.
  
  
  RESULTS
  The adipose-derived stem cells significantly suppressed T-cell proliferation. The regulatory T-cell percentages were significantly increased in the adipose-derived stem cells that were co-cultured with T cells compared with T cells alone without adipose-derived stem cell co-culture. Heme oxgenase-1 expression in concanavalin A-stimulated T cells that were co-cultured with adipose-derived stem cells revealed a significant increase compared with concanavalin A-stimulated T cells alone. Cytokine assays of the culture supernatants revealed that transforming growth factor-β and interleukin-10 were significantly increased and interferon-γ was statistically decreased in the adipose-derived stem cell-co-cultured T-cell group compared with other groups; however, blockade with a heme oxgenase-1 inhibitor (zinc protoporphyrin IX) protected against these changes.
  
  
  CONCLUSIONS
  Adipose-derived stem cells modulate T-cell proliferation and enhance regulatory T-cell expression, and this correlated with heme oxgenase-1 expression and related cytokine pathway changes.
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Ching-Ming
authors.initials: CM
authors.lastname: Chien
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Yung-Wei
authors.initials: YW
authors.lastname: Chen
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Chien-Chang
authors.initials: CC
authors.lastname: Chen
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Yi-Chia
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Shu-Hung
authors.initials: SH
authors.lastname: Huang
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Su-Shin
authors.initials: SS
authors.lastname: Lee
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Cheng-Sheng
authors.initials: CS
authors.lastname: Lai
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Sin-Daw
authors.initials: SD
authors.lastname: Lin
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Ching-Jen
authors.initials: CJ
authors.lastname: Wang
authors.affiliation: Kaohsiung, Taiwan.|From the Division of Plastic Surgery, Department of Surgery, Kaohsiung Medical University Hospital; the Departments of Plastic and Reconstructive Surgery and Orthopaedics, Kaohsiung Chang Gung Memorial Hospital; the Department of Biological Sciences, National Sun Yat-Sen University; and the Faculty of Medicine, College of Medicine, Kaohsiung Medical University.
authors.forename: Yur-Ren
authors.initials: YR
authors.lastname: Kuo
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002455:Cell Division; D018920:Coculture Techniques; D006419:Heme Oxygenase (Decyclizing) / Q000037:antagonists & inhibitors / Q000502:physiology*; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000201:enzymology / Q000502:physiology*; D011524:Protoporphyrins / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D015398:Signal Transduction / Q000276:immunology; D013154:Spleen / Q000166:cytology; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D011524:Protoporphyrins; D053773:Transforming Growth Factor beta1; D016753:Interleukin-10; C017803:zinc protoporphyrin; D007371:Interferon-gamma; D006419:Heme Oxygenase (Decyclizing); C491523:Hmox1 protein, rat