title: Novel application of ProteinChip technology exploring acute rejection markers of rat small bowel transplantation.
abstract: BACKGROUND
  Because no biomarker that reflects small bowel allograft rejection is available, we applied surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) to develop noninvasive markers required for routine diagnosis.
  
  
  METHODS
  Heterotopic small bowel transplantation (SBT) was performed in rats, and they were divided into four experimental groups: sham-operated rats (sham), syngeneic transplants (syngeneic), allogeneic transplants (allogeneic), allogeneic transplants received FK506 (allo+FK). Plasma samples were analyzed with SELDI ProteinChip arrays to detect peaks that predominated in the allogeneic model. Possible biomarkers were identified in combination with SELDI retentate chromatography mass spectrometry (RCMS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The identified protein was further analyzed by immunohistochemistry.
  
  
  RESULTS
  An increase in the level of a 14.8-kDa protein, identified as lysozyme, was observed specifically in the plasma of the allogeneic group; the levels of this protein remained unchanged in the plasma of the other groups. On the other hand, the levels of a 10.1-kDa and a 13.0-kDa protein, identified as migration inhibitory factor-related proteins (MRP), MRP-8 and MRP-14, respectively, began to increase from an early stage of acute rejection. We also observed that lysozyme-positive macrophages had strongly infiltrated the lamina propria during acute rejection.
  
  
  CONCLUSIONS
  We identified three plasma proteins-MRP-8, MRP-14, and lysozyme-that increased during small bowel allograft rejection. The identified proteins appeared to be markers for inflammation associated with allograft rejection. This proteomic approach will be useful for the identification of candidate biomarkers.
authors.affiliation: Department of Hospital Pharmacy, School of Medicine, Keio University, Tokyo, Japan.
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Yamayoshi
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Watanabe
authors.forename: Minoru
authors.initials: M
authors.lastname: Tanabe
authors.forename: Ken
authors.initials: K
authors.lastname: Hoshino
authors.forename: Koshi
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Yasuhide
authors.initials: Y
authors.lastname: Morikawa
authors.forename: Motohide
authors.initials: M
authors.lastname: Shimadzu
authors.forename: Masaki
authors.initials: M
authors.lastname: Kitajima
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Tanigawara
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers / Q000097:blood / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D004195:Disease Models, Animal; D006084:Graft Rejection / Q000276:immunology* / Q000378:metabolism*; D007150:Immunohistochemistry; D007421:Intestine, Small / Q000276:immunology* / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D008297:Male; D008969:Molecular Sequence Data; D009113:Muramidase / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
chemical_list: D015415:Biomarkers; D001798:Blood Proteins; D009113:Muramidase