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영어

Xenotransplantation has been presented as a strategy to meet the imbalance between organ donors and recipients. However, the immune suitability of organs between the animal organs and human bodies do not match, therefore, hyperacute-rejection (HAR) and antibody-dependent cellular cytotoxicity (ADCC), cell-mediated immune rejection arises from transplanting animal organ becomes necrosis appeared in a short time. Recently, various genetic engineering has produced GTKO pigs that knockout alpha- 1,3-gal genes known to be the cause of hyperacute rejection, In addition, various immunosuppressive studies are underway. The ADAM10 and 17 genes are proteins belonging to a disintegrin metalloprotease (ADAM) family and mainly function as sheddase. In normal cells, ADAM10 and ADAM17 has cut off EGH ligands to produce growth hormone, or TGFa is activated to inhibit viral infection. But, in tumor cells, MHC Class I-related proteins A, B and other ligands expressed on the cell surface are converted into soluble forms by ADAM10, 17 proteins and immunosuppressive reactions are performed for survival of tumor cells. The aim of this study was to investigate whether ADAM10 and 17 genes could be used to mimic immune-evasion mechanisms of tumor cells into normal cells to reduce cell-mediated rejection. In this experiment, the ADAM 10, 17 gene was obtained miniature pigs and amplified by PCR, inserted into pcDNA3.1 vector, and introduced Porcine Fetal fibroblast (PFF). Transgene cells selected by geneticin (500 ug/mL) treatment for 2 weeks were used target cell for cytotoxicity experiments. NK92MI cells (ATCC, CR2408) were used for NK cell cytotoxicity assay, and CD8+ T cells were separated by Ficoll gradient and stimulated with PMA (25 ng/mL) and IL-2 (50 U/mL) for 2 weeks. The results showed that the cytotoxic inhibitory effect of ADAM10, 17 on CD8+ T cells was greater than NK cells. Further studies will need to evaluate the soluble forms of NKG2DL that are involved in the immune response of NK cells and CD8+ T cells.