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Rezeptorvermittelter Gentransfer Anwendung in der Tumorimmunotherapie?

  • E. Wagner
Conference paper

Zusammenfassung

Genkonstrukte, die mit Polylysin-konjugierten Liganden wie Transferrin, EGF, oder Anti-CD 3-Antikörpern komplexiert sind, werden über Rezeptor-vermittelte Endozytose in Säugerzellen eingeschleust. Für einen effizienten Gentransfer müssen die Genkonstrukte aus der vesikulären Struktur der Endosomen in das Zytoplasma und weiter in den Zellkern transportiert werden. Als Endosomen-destabilisierende Komponenten wurden synthetische, von Influenzavirus- oder Rhinovirussequenzen abgeleitete Peptide oder ganze, inaktivierte Adenoviren eingesetzt. Als mögliche Anwendung des Gentransfersystems wird die Herstellung einer autologen, genmodifizierten Tumorvakzine zur Immunotherapie vorgestellt.

Summary

Gene constructs have been complexed with polylysine-conjugated ligands (such as transferrin, EGF, or anti-CD3 antibodies) for receptor-mediated uptake into endosomes. Accumulation of these complexes in intracellular vesicles however strongly reduces the efficiency of gene transfer. Viruses have acquired mechanisms to release their genome from endosomes into the cytoplasm. The endosomal acidification process specifically activates viral surface proteins that trigger desta-bilization of the endosomal membrane. This has led to the development of virus-like gene transfer complexes consisting of DNA complexed with a cell-binding ligand and polylysine-conjugated, endosomedisruption agents (such as defective adenoviruses, or small synthetic peptides derived from sequences of the influenza virus hemagglutinin or the rhinovirus VP-1 protein) which allow cytoplasmic entry of the DNA. DNA complexes linked to psoralen/UV-inactivated adenoviruses have been applied for the expression of high levels of cytokine genes in primary human cancer cells. The utility of the gene transfer system for the generation of gene-modified cancer vaccines is discussed.

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Literatur

  1. 1.
    Cotten M, Wagner E (1993) Non-viral approaches to gene therapy. Current Op Biotech 4:705–710.CrossRefGoogle Scholar
  2. 2.
    Cotten M, Wagner E, Birnstiel ML (1993) Receptor mediated transport of DNA into eukariotic cells. Meth Enzymol 217:618–644.PubMedCrossRefGoogle Scholar
  3. 3.
    Cotten M, Wagner E, Zatloukal K, Birnstiel ML (1993) Chicken adenovirus (CELO virus) particles augment receptor-mediated DNA delivery to mammalian cells and yield exceptional levels of stable transformants. J Virol 67:3777–3785.PubMedGoogle Scholar
  4. 4.
    Curiel TI, Cook DR, Bogedain C, Jilg W, Harrison GS, Cotten M, Curiel DT, Wanger E (1994) Foreign gene expression in Epstein-Barr virus transformed human B cells. Virology 198:577–585.PubMedCrossRefGoogle Scholar
  5. 5.
    Plank C, Zatloukal K, Cotten M, Mechtler K, Wagner E (1992) Gene transfer into hepatocytes using asialoglycoprotein receptor mediated endocytosis of DNA complexed with an artificial tetra-antennary galactose ligand. Bioconjugate Chem 3:533–539.CrossRefGoogle Scholar
  6. 6.
    Plank C, Oberhauser B, Mechtler K, Koch C, Wagner E (1994) The influence of endosome-disruptive peptides on gene transfer using synthetic virus-like gene transfer systems. J Biol Chem 269:12918–12924.PubMedGoogle Scholar
  7. 7.
    Wagner E, Zatloukal K, Cotten M Kirlappos H, Mechtler K, Curiel DT, Birnstiel ML (1992) Coupling of adenovirus to transferrin polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes. Proc Natl Acad Sci USA 89:6099–6103.PubMedCrossRefGoogle Scholar
  8. 8.
    Wagner E, Curiel D, Cotten M (1994) Delivery of drugs, proteins and genes into cells using transferrin as a ligand for receptor-mediated endocytosis. Adv Drug Del Rev 14:113–136.CrossRefGoogle Scholar
  9. 9.
    Zatloukal K, Schmidt W, Cotten M, Wagner E, Stingl G, Birnstiel ML (1993) Somatic gene therapy for cancer: the utility of transferrinfection in generating “tumor vaccines”. Gene 135:199–207.PubMedCrossRefGoogle Scholar
  10. 10.
    Zatloukal K, Cotten M, Berger M, Schmidt W, Wagner E, Birnstiel ML (1994) In vivo production of human factor VIII in mice after intrasplenic implantation of primary fibroblasts transfected by receptor-mediated, adenovirus-augmented gene delivery. Proc Natl Acad Sci USA 91:5148–5152.PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1995

Authors and Affiliations

  • E. Wagner

There are no affiliations available

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