title: DNA microarray-based gene expression profiling in porcine keratocytes and corneal endothelial cells and comparative analysis associated with xeno-related rejection.
abstract: 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.
authors.affiliation: Seoul Artificial Eye Center, Seoul National University Hospital Clinical Research Institute, Seoul, Korea.
authors.forename: Mee Kum
authors.initials: MK
authors.lastname: Kim
authors.forename: Joo Youn
authors.initials: JY
authors.lastname: Oh
authors.forename: Jung Hwa
authors.initials: JH
authors.lastname: Ko
authors.forename: Hyun Ju
authors.initials: HJ
authors.lastname: Lee
authors.forename: Jin Ho
authors.initials: JH
authors.lastname: Jung
authors.forename: Won Ryang
authors.initials: WR
authors.lastname: Wee
authors.forename: Jin Hak
authors.initials: JH
authors.lastname: Lee
authors.forename: Chung-Gyu
authors.initials: CG
authors.lastname: Park
authors.forename: Sang Joon
authors.initials: SJ
authors.lastname: Kim
authors.forename: Curie
authors.initials: C
authors.lastname: Ahn
authors.forename: Seung-Jun
authors.initials: SJ
authors.lastname: Kim
authors.forename: Seung Yong
authors.initials: SY
authors.lastname: Hwang
mesh_terms: 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
chemical_list: D018932:Chemokine CCL2; D015234:HLA-A Antigens; D015923:Complement C1r