In vivo liposome-mediated transfection of HLA-DR alpha-chain gene into pig hearts

被引:2
|
作者
Aleksic, I [1 ]
Ren, M [1 ]
Popov, A [1 ]
Freimark, D [1 ]
Blanche, C [1 ]
Czer, L [1 ]
Trento, A [1 ]
Barath, P [1 ]
机构
[1] Univ Gottingen, Dept Thorac & Cardiovasc Surg, D-37075 Gottingen, Germany
关键词
liposomes; gene transfer; cardiac xenotransplantation;
D O I
10.1016/S1010-7940(97)00247-9
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Scarcity of suitable donor organs remains a major problem for organ transplantation. Transfer of recipient HLA-genes into animal donor-organs during harvest could induce graft-tolerance without suppressing the recipient immune system. Objective: This pilot study aimed to test the feasibility of an in vivo gene transfer into pig hearts by intracoronary infusion of DNA:liposome-complexes and to detect the gene product by immunuhistochemistry. Methods: The pcDV1-pL2-vector, containing the basesequence for HLA-DR alpha-chain in plasmids (1.3 kb) was selected. The plasmids were isolated with ethidiumbromide and incubated with lipofectin(R) in a 1:3-ratio for 10 min. The DNA:lipofectin(R)-complex was diluted to 10 cc with physiologic saline and delivered into the left anterior descending artery of 6 farm pigs over 10 min. As a control within the same animal, the same amount of lipofectin(R) alone was infused into the first diagonal branch. Three pigs were sacrificed after 24 h, the other 3 after 48 h. Delivery of DNA:liposome-complexes was detected by oil red 0 staining, expression of HLA-DR alpha-chain-antigen with a monoclonal anti-HLA-DR alpha-antibody. Results: Transfection of the HLA-class-II DR-alpha-chain occurred in endothelial cells. Infiltrating cells around capillaries stained positively for HLA-DR-alpha. These infiltrating cells were negative for the pan B- and the pan T-cell-marker L26 and UCHL-1. There was no transfection and hypercellularity in the myocardium around the first diagonal branch. Conclusion: In vivo intracoronary infusion of the HLA-DR alpha-chain-DNA:lipofectin(R)-complex leads to expression of the corresponding antigen on pig endothelium for 48 h. The infiltrating cells require further characterization. (C) 1997 Elsevier Science B.V.
引用
收藏
页码:792 / 797
页数:6
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