Anne Behnecke1, Wei Li, Ling Chen, Andrew Saxon, Ke Zhang. 1. Hart and Louise Lyon Immunology Laboratory, Section of Clinical Immunology/Allergy, Division of Pulmonary, Critical Care and Clinical Immunology/Allergy, Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
Abstract
BACKGROUND: Treatment of IgE-mediated food allergy with standard protein-based allergen immunotherapy has proved both unsuccessful and hazardous. Allergen gene vaccination represents a promising alternative, but difficulties in gene targeting and expression in antigen-presenting cells represent a major limitation for efficient gene vaccination. OBJECTIVE: We sought to construct a genetically engineered human epsilon-polylysine (EPL) fusion protein that binds allergen gene expression systems and targets the gene vaccine complex to antigen-presenting cells through the interaction of EPL and the high-affinity receptor for IgE for efficient allergen gene vaccination. METHODS: Genetic engineering was used to design and produce the EPL fusion gene, consisting of the human CHepsilon2-4 linked to 55 lysine residues, and the conventional approaches were used to characterize the biologic features of EPL. RESULTS: EPL was assembled as functional dimers and capable of binding DNA plasmids in both an EPL protein and plasmid DNA concentration-dependent manner. EPL targeted plasmid DNA to the high-affinity receptor for IgE on cell surfaces and increased the model gene uptake/expression. The EPL-DNA complexes were shown not to trigger mast cell degranulation. CONCLUSION: EPL is able to function as a gene carrier system to target allergen gene to the high-affinity receptor for IgE-expressing cells through ligand receptor-mediated interactions.
BACKGROUND: Treatment of IgE-mediated food allergy with standard protein-based allergen immunotherapy has proved both unsuccessful and hazardous. Allergen gene vaccination represents a promising alternative, but difficulties in gene targeting and expression in antigen-presenting cells represent a major limitation for efficient gene vaccination. OBJECTIVE: We sought to construct a genetically engineered humanepsilon-polylysine (EPL) fusion protein that binds allergen gene expression systems and targets the gene vaccine complex to antigen-presenting cells through the interaction of EPL and the high-affinity receptor for IgE for efficient allergen gene vaccination. METHODS: Genetic engineering was used to design and produce the EPL fusion gene, consisting of the human CHepsilon2-4 linked to 55 lysine residues, and the conventional approaches were used to characterize the biologic features of EPL. RESULTS: EPL was assembled as functional dimers and capable of binding DNA plasmids in both an EPL protein and plasmid DNA concentration-dependent manner. EPL targeted plasmid DNA to the high-affinity receptor for IgE on cell surfaces and increased the model gene uptake/expression. The EPL-DNA complexes were shown not to trigger mast cell degranulation. CONCLUSION: EPL is able to function as a gene carrier system to target allergen gene to the high-affinity receptor for IgE-expressing cells through ligand receptor-mediated interactions.
Authors: R Ross; S Sudowe; J Beisner; X-L Ross; I Ludwig-Portugall; J Steitz; T Tüting; J Knop; A B Reske-Kunz Journal: Gene Ther Date: 2003-06 Impact factor: 5.250
Authors: D Maurer; E Fiebiger; B Reininger; C Ebner; P Petzelbauer; G P Shi; H A Chapman; G Stingl Journal: J Immunol Date: 1998-09-15 Impact factor: 5.422