Experimental study for the immunological tolerance using orthotopic liver transplantation using mice

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In mouse liver transplantation(OMLT), immunological tolerance is induced spontaneously across MHC barrier. To identify specific protein and gene for inducing tolerance, differential display and DNA tip analysis performed using mouse liver that acquired tolerance. As the result, 40kDa and 75kDa protein was detected. On the other hand, several genes specific for tolerance are identified using DNA tip, for example secretory leukocyte protease inhibitor, lymphotoxin B, rpt-1, CD24, which are related to immunological response.Flowcytemetry revealed the existence of the double-positive cells which expressed both MHC class I antigen for donor and recipient in the grafted liver. Analysis of surface antigens showed the double-positive cells possessed the characteristics of the premature dendritic cells of the recipient origin. RT-PCR using specific primers for donor and recipient antigen demonstrated that it was constantly generated in the grafted liver. These data suggested that the double-positive cells were possibly the recipient type premature dendritic cells which trapped the donor type soluble MHC class I antigen to their surface.And to make the soluble MHC class I antigen, we constructed the cellular surface domain, α1, α2, α3, and β2 microgloblin using pichia vector, and transformed to Pichia pastories. Now the protein has been purified, after that, it will be administrated naive mice and examine the ability to induce tolerance.