Influenza M2 envelope protein augments avian influenza hemagglutinin pseudotyping of lentiviral vectors

• Published in: Gene therapy Vol. 13; no. 8; pp. 715 - 724
• Main Authors: MCKAY, T, PATEL, M, PICKLES, R. J, JOHNSON, L. G, OLSEN, J. C
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.04.2006
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Applied cell therapy and gene therapy; Avian flu; Avian influenza; Biological and medical sciences; Biotechnology; Channel gating; Cystic fibrosis; Drug resistance; Epithelial cells; Equine infectious anemia; Exo-a-sialidase; Expression vectors; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Gene therapy; Gene transfer; Genetic aspects; Genetic Engineering; Genetic Therapy - methods; Genetic transformation; Genetic vectors; Genetic Vectors - administration & dosage; Genetic Vectors - genetics; Genotype; Health. Pharmaceutical industry; Hemagglutinins; Hemagglutinins, Viral - genetics; HIV; HIV-1 - genetics; Human immunodeficiency virus; Human viral diseases; Humans; Industrial applications and implications. Economical aspects; Infectious Anemia Virus, Equine - genetics; Infectious diseases; Influenza; Influenza A virus - genetics; Lentivirus - genetics; Lung diseases; Lung Diseases - therapy; Medical sciences; Mice; Mice, Inbred C57BL; Plague; Producer cells; Protein research; Proteins; Rats; Resistant mutant; Respiratory diseases; Rodents; Throat; Trachea - virology; Transduction, Genetic - methods; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Viral diseases; Viral diseases of the respiratory system and ent viral diseases; Viral envelope proteins; Viral Matrix Proteins - genetics; Viruses; United States; Virus; Avian influenza; airway epithelia; Hemagglutinin; Retroviridae; influenza virus; Envelope; Gene therapy; Lentivirus; Vector; Protein; pseudotype
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3302715

Lentivirus-based gene transfer has the potential to efficiently deliver DNA-based therapies into non-dividing epithelial cells of the airway for the treatment of lung diseases such as cystic fibrosis. However, significant barriers both to lung-specific gene transfer and to production of lentivirus vectors must be overcome before these vectors can be routinely used for applications to the lung. In this study, we investigated whether the ability to produce lentiviral vectors pseudotyped with fowl plague virus hemagglutinin (HA) could be improved by co-expression of influenza virus M2 in vector-producing cells. We found that M2 expression led to a 10-30-fold increase in production of HA-pseudotyped lentivirus vectors based upon equine infectious anemia virus (EIAV) or human immunodeficiency virus type 1 (HIV-1). Experiments using the M2 inhibitor amantadine and a drug-resistant mutant of M2 established that the ion channel activity of M2 was important for M2-dependent augmentation of vector production. Furthermore, the neuraminidase activity necessary for particle release from producer cells could also be incorporated into producer cells by co-expression of influenza NA cDNA. Lentiviral vectors pseudotyped with influenza envelope proteins were able to efficiently transduce via the apical membrane of polarized mouse tracheal cultures in vitro as well as mouse tracheal epithelia in vivo.