Rescue of retroviral envelope fusion deficiencies by cationic liposomes

Abstract

Background Retroviral particles that are inappropriately enveloped can transduce target cells if pre-associated with cationic liposomes. This study optimises and addresses the mechanism of liposome-enhanced gene delivery, and explores the potential for such agents to compensate for fusion deficiency associated with chimaeric envelope proteins. Methods Particles bearing wild-type, chimaeric or no envelope proteins were complexed with DOTAP or DC-Chol/DOPE cationic liposomes and added to target cells for various times. Particle binding was determined by detection of cell-associated capsid protein and infectivity was measured histochemically. Results Stable association of cationic liposomes with retrovirus particles significantly enhanced their binding rate to target cells in proportion to the increase of transduction kinetics for infectious virus. Binding of virus was equivalent with or without envelope protein and/or virus receptor, indicating that a non-specific interaction precedes receptor recognition. Non-infectious combinations were rescued by the intrinsic fusogenicity of the cationic liposomes, which enabled entry of the viral core, but left subsequent events unaltered. The optimised transduction rate with non-enveloped particles and DOTAP approached that of amphotropic-enveloped virus in some cases, although the effect was target-cell-dependent. DC-Chol/DOPE was less potent at direct fusion but was able to enhance 600-fold the receptor-dependent action of chimaeric envelopes that were deficient in fusion by virtue of the addition of targeting domains. Conclusions These data have implications for the development of retroviral vector targeting strategies from the perspectives of the specificity of target cell interaction and compensating for chimaeric envelope fusion deficiency. Copyright (C) 2002 John Wiley Sons, Ltd.