DNA microarray-based gene expression profiling in porcine keratocytes and corneal endothelial cells and comparative analysis associated with xeno-related rejection.
Porcine to rat corneal xenotransplantation resulted in severe inflammation and rejection of the corneal stroma, whereas an allograft showed mainly endothelial cell-associated rejection. We, therefore, investigated and compared the gene expression between porcine keratocytes and corneal endothelial cells. RNA was isolated from primary cultured porcine or human keratocytes and porcine corneal endothelial cells. Gene expression was comparatively analyzed after normalization with microarray method using Platinum pig 13 K oligo chip (GenoCheck Co., Ltd., Ansan, Korea). Real-time polymerase chain reaction (PCR) was performed for C1R, CCL2, CXCL6, and HLA-A in porcine keratocytes and corneal endothelial cells. As a result, upregulated expression more than 2 folds was observed in 1,162 genes of porcine keratocytes versus porcine endothelial cells. Among the immune-regulatory genes, SEMA3C, CCL2, CXCL6, F3, HLA-A, CD97, IFI30, C1R, and G1P3 were highly expressed in porcine keratocytes, compared to porcine corneal endothelial cells or human keratocytes. When measured by real-time PCR, the expression of C1R, CCL2, and HLA-A was higher in porcine keratocytes compared to that in porcine corneal endothelial cells. In conclusion, the increased expression of C1R, CCL2, and HLA-A genes in porcine keratocytes might be responsible for the stromal rejection observed in a porcine to rat corneal xenotransplantation.
Kim
Mee Kum
MK
Seoul Artificial Eye Center, Seoul National University Hospital Clinical Research Institute, Seoul, Korea.
Oh
Joo Youn
JY
Ko
Jung Hwa
JH
Lee
Hyun Ju
HJ
Jung
Jin Ho
JH
Wee
Won Ryang
WR
Lee
Jin Hak
JH
Park
Chung-Gyu
CG
Kim
Sang Joon
SJ
Ahn
Curie
C
Kim
Seung-Jun
SJ
Hwang
Seung Yong
SY
D000818:Animals; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000378:metabolism; D015923:Complement C1r / Q000378:metabolism; D016039:Corneal Transplantation / Q000276:immunology* / Q000473:pathology; D004728:Endothelium, Corneal / Q000378:metabolism* / Q000473:pathology; D020869:Gene Expression Profiling; D006084:Graft Rejection / Q000276:immunology* / Q000473:pathology; D015234:HLA-A Antigens / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D014183:Transplantation, Heterologous; D015854:Up-Regulation
D018932:Chemokine CCL2; D015234:HLA-A Antigens; D015923:Complement C1r