Novel application of ProteinChip technology exploring acute rejection markers of rat small bowel transplantation.
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.
Yamayoshi
Yasuko
Y
Department of Hospital Pharmacy, School of Medicine, Keio University, Tokyo, Japan.
Watanabe
Toshihiko
T
Tanabe
Minoru
M
Hoshino
Ken
K
Matsumoto
Koshi
K
Morikawa
Yasuhide
Y
Shimadzu
Motohide
M
Kitajima
Masaki
M
Tanigawara
Yusuke
Y
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
D015415:Biomarkers; D001798:Blood Proteins; D009113:Muramidase