title: Syngeneic adipose-derived stem cells with short-term immunosuppression induce vascularized composite allotransplantation tolerance in rats.
abstract: OBJECTIVE
  A clinically applicable tolerance induction regimen that removes the requirement for lifelong immunosuppression would benefit recipients of vascularized composite allotransplantation (VCA). We characterized the immunomodulatory properties of syngeneic (derived from the recipient strain) adipocyte-derived stem cells (ADSCs) and investigated their potential to induce VCA tolerance in rats.
  
  
  METHODS
  ADSCs were isolated from Lewis (LEW, RT1A(l)) rats; their immunomodulatory properties were evaluated by means of mixed lymphocyte reactions in vitro and VCAs in vivo across a full major histocompatibility complex mismatch with the use of Brown-Norway (BN, RT1A(n)) donor rats. Two control and four experimental groups were designed to evaluate treatment effects of ADSCs and transient immunosuppressants (anti-lymphocyte globulin, cyclosporine) with or without low-dose (200 cGy) total body irradiation. Flow cytometry was performed to quantify levels of circulating CD4(+)CD25(+)FoxP3(+) regulatory T cells (Tregs).
  
  
  RESULTS
  Cultured syngeneic ADSCs exhibited CD90.1(+)CD29(+)CD73(+)CD45(-)CD79a(-)CD11b/c(-) phenotype and the plasticity to differentiate to adipocytes and osteocytes. ADSCs dramatically suppressed proliferation of LEW splenocytes against BN antigen and mitogen, respectively, in a dose-dependent fashion, culminating in abrogation of allo- and mitogen-stimulated proliferation at the highest concentration tested. Accordingly, one infusion of syngeneic ADSCs markedly prolonged VCA survival in LEW recipients treated with transient immunosuppression; of these, 66% developed tolerance. Total body irradiation provided no additional VCA survival benefit. An important role for Tregs in tolerance induction/maintenance was suggested in vivo and in vitro.
  
  
  CONCLUSIONS
  Treatment comprising syngeneic ADSCs and transient immunosuppression (i) increased levels of circulating Tregs and (ii) induced tolerance in 66% of recipients of major histocompatibility complex-mismatched VCAs.
authors.affiliation: Center for Vascularized Composite Allotransplantation, Chang Gung Memorial Hospital, Gueishan, Taiwan; Department of Medical Research and Development Linkou Branch, Chang Gung Medical Foundation, Taoyuan, Gueishan, Taiwan.
authors.forename: Hui-Yun
authors.initials: HY
authors.lastname: Cheng
authors.affiliation: Former Microsurgery Fellow, Chang Gung Memorial Hospital; Regional Oncological Institute, University of Medicine and Pharmacy. "Grigore T. Popa," Iasi, România.
authors.forename: Nicolae
authors.initials: N
authors.lastname: Ghetu
authors.affiliation: Division of Plastic and Reconstructive Surgery, Tzu Chi General Hospital at Taipei, New Taipei, Taiwan.
authors.forename: Wei-Chao
authors.initials: WC
authors.lastname: Huang
authors.affiliation: Center for Vascularized Composite Allotransplantation, Chang Gung Memorial Hospital, Gueishan, Taiwan.
authors.forename: Yen-Ling
authors.initials: YL
authors.lastname: Wang
authors.affiliation: Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Gueishan, Taiwan.
authors.forename: Christopher Glenn
authors.initials: CG
authors.lastname: Wallace
authors.affiliation: School of Medicine, Chang Gung University, Gueishan, Taiwan.
authors.forename: Chih-Jen
authors.initials: CJ
authors.lastname: Wen
authors.affiliation: Graduate Institute of Biomedical Sciences, Chang Gung University, Gueishan, Taiwan.
authors.forename: Hung-Chang
authors.initials: HC
authors.lastname: Chen
authors.affiliation: Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Gueishan, Taiwan.
authors.forename: Ling-Yi
authors.initials: LY
authors.lastname: Shih
authors.affiliation: School of Medicine, Chang Gung University, Gueishan, Taiwan.
authors.forename: Chih-Fan
authors.initials: CF
authors.lastname: Lin
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, Hsinchu, Taiwan.
authors.forename: Shiaw-Min
authors.initials: SM
authors.lastname: Hwang
authors.affiliation: Graduate Institute of Cancer Biology and Drug Discovery and Center of Excellence for Cancer Research, Taipei Medical University, Taipei, Taiwan; R&D Division, Vectorite Biomedica Inc, Taipei, Taiwan. Electronic address: liaosk@h.tmu.edu.tw.
authors.forename: Shuen-Kuei
authors.initials: SK
authors.lastname: Liao
authors.affiliation: Center for Vascularized Composite Allotransplantation, Chang Gung Memorial Hospital, Gueishan, Taiwan; Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Gueishan, Taiwan; School of Medicine, Chang Gung University, Gueishan, Taiwan. Electronic address: fuchanwei@gmail.com.
authors.forename: Fu-Chan
authors.initials: FC
authors.lastname: Wei
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D006649:Histocompatibility Antigens / Q000378:metabolism; D006801:Humans; D007165:Immunosuppression Therapy; D007166:Immunosuppressive Agents / Q000008:administration & dosage; D007519:Isoantigens / Q000276:immunology; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D013234:Stem Cells / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D023001:Transplantation Tolerance; D063986:Vascularized Composite Allotransplantation
chemical_list: D006649:Histocompatibility Antigens; D007166:Immunosuppressive Agents; D007519:Isoantigens
keywords: adipose-derived stem cell; allotransplantation; regulatory T cell; tolerance; vascularized composite