| Co-Investigator(Kenkyū-buntansha) |
ENOSAWA Shin Deputy Director, Dept. of Exp. Surgery, National Children's Medical Research Center, 小児医療研究センター・実験外科生体工学部, 室長 (40232962)
OKUYAMA Torayuki Deputy Director, Dept. of genetics, National Children's Medical Research Center, 小児医療研究センター・先天異常研究部, 室長 (40177192)
李 小康 国立小児病院, 小児医療研究センター・実験外科生体工学部, 研究員
SUZUKE Seiichi Director, Dept. of Exp. Surgery, National Children's Medical Research Center, 小児医療研究センター・実験外科生体工学部, 部長 (00111386)
LI Xiao-Kang Post-doctory fellow, National Children's Medical Research Center
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| Research Abstract |
The preventive effect of Fas-1igand (FasL) expression on allogeneic liver transplantation was investigated using two independent exogenous gene transfer and expression methods. First, HVJ-FasL, an HVJ-liposome incorporating pCAGGSFasL, containing an expression unit of rat FasL CDNA under the control of chiken β-actin promoter and CMV enhancer, was generated and injected into DA rats via portal vein. Second, a recombinant adenovirus AxCALNFasL was constructed in order to transduce a gene for rat FasL, requiring co-expression of Cre recombinase for its expressing. The gene expression level was studied X-gal staining, showed about 12% and 20% positive cells in liver sections obtained from the control HVJ-pCAGGSLZ and AxCALNLZ transferred livers, respectively. Expression of FasL in the gene transferred liver with HVJ-FasL and AxCALNFasL was confirmed by RT-PCR and Westem blotting. Median survival times the Lewis rats transplanted DA Iiver pretreated with HVJ-FasL and AxCALNFasL were 18 day
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s ( n= 8 ), and 22 days( n=4), rsespectively. On the other hand, median survival time of Lewis rats transplanted DA liver pretreated with control HVJ-liposome or adenovirus was only 11 days ( n=8 ), suggesting that the FasL gene expression on allograft contributed to the control of immune response in allogeneic liver transplantation. We conclude that an appropriate expression level of FasL in donor liver probably acts to induce apoptosis to recipient activated T cells expressing a large amount of Fas on their cell surface, which resulted in the regulation of immune response generated in allogeneic liver transplantation.
Using a simple but clear model of liver transplantation in congenic mice (MRL-lpr/lpr (lpr) and MRL+/+ (wild type) mice), we investigated how inflammatory reactions were induced in the liver after FasL-transfection. Adenovirus vector expressing FasL in the Fas-negative liver thus lead to lethal hepatitis in the Fas-positive syngeneic recipients but not in the Fas-negative animals, which clearly indicated that FasL, apart from the direct interaction with Fas antigens, may have the potent ability to accumulate in the liver and activate the Fas-positive inflammatory cells. The present data demonstrating the role of FasL in inflammation. This is important for generating a new strategy to prevent hepatitis as well as for understanding the role of the Fas/FasL interaction in the pathophysiology of hepatitis . Less
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