In vivo liposome-mediated transfection of HLA-DR alpha-chain gene into pig hearts.
Eur J Cardiothorac Surg
; 12(5): 792-7, 1997 Nov.
Article
em En
| MEDLINE
| ID: mdl-9458153
ABSTRACT
UNLABELLED 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 DNAliposome-complexes and to detect the gene product by immunohistochemistry.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 in a 13-ratio for 10 min. The DNAlipofectin-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 alone was infused into the first diagonal branch. Three pigs were sacrificed after 24 h, the other 3 after 48 h. Delivery of DNAliposome-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.CONCLUSIONS:
In vivo intracoronary infusion of the HLA-DR alpha-chain-DNAlipofectin-complex leads to expression of the corresponding antigen on pig endothelium for 48 h. The infiltrating cells require further characterization.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fosfatidiletanolaminas
/
Transfecção
/
Antígenos HLA-DR
/
Miocárdio
Limite:
Animals
Idioma:
En
Revista:
Eur J Cardiothorac Surg
Assunto da revista:
CARDIOLOGIA
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Alemanha