title: Proteomic analysis in serum of rat hind-limb allograft tolerance induced by immunosuppressive therapy with adipose-derived stem cells.
abstract: BACKGROUND
  Adipose-derived stem cells combined with transient immunosuppression prolonged vascularized composite tissue allotransplant survival and induced immune tolerance in a rodent hind-limb model. The authors investigated serum proteins in the adipose-derived stem cell tolerance group and control group using proteomic study.
  
  
  METHODS
  An orthotopic hind-limb model from Brown-Norway to Lewis rats was used. The control group received no treatment. Rats in the tolerance group received combined treatments of short-term cyclosporine A, antilymphocyte serum, and multiple rounds of adipose-derived stem cells. Serum samples were analyzed. Spots of interest were subjected to in-gel trypsin digestion and matrix-assisted laser desorption ionization time-of-flight mass spectrometry to elucidate the peptide mass fingerprints. The mass spectrometric characteristics of the identified proteins were analyzed. Immunohistochemical analysis of transplanted tissue and enzyme-linked immunosorbent assay of serum were validated.
  
  
  RESULTS
  Rats in the tolerance group had significantly higher amounts of β2-glycoprotein, α1-macroglobulin, rat-albumin, and vitamin D-binding protein, and significantly lower levels of haptoglobin compared with controls. Immunohistochemical staining of the alloskin indicated similar effects, such as up-regulated vitamin D-binding protein and down-regulated haptoglobin in the tolerance group compared with rejection controls (p < 0.05). Enzyme-linked immunosorbent assay revealed that vitamin D-binding protein was statistically increased (p < 0.05) and haptoglobin expression was significantly decreased (p < 0.01) in the tolerance group compared with the controls.
  
  
  CONCLUSIONS
  There were significant differences in the serum proteomics between the tolerance and control groups. Down-regulated haptoglobin and up-regulated vitamin D-binding protein are involved in adipose-derived stem cell-induced immune tolerance and allotransplant survival.
authors.affiliation: Kaohsiung and Tao-Yuan, Taiwan From the Department of Plastic and Reconstructive Surgery, the Department of Liver Transplantation Program, and the Department of Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine; the Department of Biological Sciences, National Sun Yat-sen University; and the Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University.
authors.forename: Yur-Ren
authors.initials: YR
authors.lastname: Kuo
authors.forename: Chien-Chang
authors.initials: CC
authors.lastname: Chen
authors.forename: Shigeru
authors.initials: S
authors.lastname: Goto
authors.forename: Yu-Ting
authors.initials: YT
authors.lastname: Huang
authors.forename: Chia-Chun
authors.initials: CC
authors.lastname: Tsai
authors.forename: Ming-Yu
authors.initials: MY
authors.lastname: Yang
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D000961:Antilymphocyte Serum / Q000494:pharmacology / Q000627:therapeutic use; D003131:Combined Modality Therapy; D016572:Cyclosporine / Q000494:pharmacology / Q000627:therapeutic use; D004359:Drug Therapy, Combination; D004797:Enzyme-Linked Immunosorbent Assay; D006084:Graft Rejection / Q000097:blood / Q000276:immunology / Q000517:prevention & control*; D006614:Hindlimb / Q000637:transplantation*; D007150:Immunohistochemistry; D007166:Immunosuppressive Agents / Q000494:pharmacology / Q000627:therapeutic use*; D008297:Male; D013058:Mass Spectrometry; D045164:Mesenchymal Stem Cell Transplantation; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D051381:Rats; D011914:Rats, Inbred BN; D011917:Rats, Inbred Lew; D023001:Transplantation Tolerance / Q000187:drug effects / Q000502:physiology; D016896:Treatment Outcome; D063986:Vascularized Composite Allotransplantation
chemical_list: D000961:Antilymphocyte Serum; D007166:Immunosuppressive Agents; D020543:Proteome; D016572:Cyclosporine