In vivo liposome-mediated transfection of HLA-DR alpha-chain gene into pig hearts. 1997

I Aleksic, and M Ren, and A Popov, and D Freimark, and C Blanche, and L Czer, and A Trento, and P Barath
Department of Thoracic and Cardiovascular Surgery, Georg August University, Göttingen, Germany.

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 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 1:3-ratio for 10 min. The DNA:lipofectin-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 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. CONCLUSIONS In vivo intracoronary infusion of the HLA-DR alpha-chain-DNA:lipofectin-complex leads to expression of the corresponding antigen on pig endothelium for 48 h. The infiltrating cells require further characterization.

UI MeSH Term Description Entries
D007150 Immunohistochemistry Histochemical localization of immunoreactive substances using labeled antibodies as reagents. Immunocytochemistry,Immunogold Techniques,Immunogold-Silver Techniques,Immunohistocytochemistry,Immunolabeling Techniques,Immunogold Technics,Immunogold-Silver Technics,Immunolabeling Technics,Immunogold Silver Technics,Immunogold Silver Techniques,Immunogold Technic,Immunogold Technique,Immunogold-Silver Technic,Immunogold-Silver Technique,Immunolabeling Technic,Immunolabeling Technique,Technic, Immunogold,Technic, Immunogold-Silver,Technic, Immunolabeling,Technics, Immunogold,Technics, Immunogold-Silver,Technics, Immunolabeling,Technique, Immunogold,Technique, Immunogold-Silver,Technique, Immunolabeling,Techniques, Immunogold,Techniques, Immunogold-Silver,Techniques, Immunolabeling
D009206 Myocardium The muscle tissue of the HEART. It is composed of striated, involuntary muscle cells (MYOCYTES, CARDIAC) connected to form the contractile pump to generate blood flow. Muscle, Cardiac,Muscle, Heart,Cardiac Muscle,Myocardia,Cardiac Muscles,Heart Muscle,Heart Muscles,Muscles, Cardiac,Muscles, Heart
D010714 Phosphatidylethanolamines Derivatives of phosphatidic acids in which the phosphoric acid is bound in ester linkage to an ethanolamine moiety. Complete hydrolysis yields 1 mole of glycerol, phosphoric acid and ethanolamine and 2 moles of fatty acids. Cephalin,Cephalins,Ethanolamine Phosphoglyceride,Ethanolamine Phosphoglycerides,Ethanolamineglycerophospholipids,Phosphoglyceride, Ethanolamine,Phosphoglycerides, Ethanolamine
D010865 Pilot Projects Small-scale tests of methods and procedures to be used on a larger scale if the pilot study demonstrates that these methods and procedures can work. Pilot Studies,Pilot Study,Pilot Project,Project, Pilot,Projects, Pilot,Studies, Pilot,Study, Pilot
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D006684 HLA-DR Antigens A subclass of HLA-D antigens that consist of alpha and beta chains. The inheritance of HLA-DR antigens differs from that of the HLA-DQ ANTIGENS and HLA-DP ANTIGENS. HLA-DR,Antigens, HLA-DR,HLA DR Antigens
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013552 Swine Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA). Phacochoerus,Pigs,Suidae,Warthogs,Wart Hogs,Hog, Wart,Hogs, Wart,Wart Hog
D014162 Transfection The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES. Transfections

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