Transduction by adeno-associated virus vectors in the rabbit airway: efficiency, persistence, and readministration

The ability of recombinant adeno-associated virus (AAV) vectors to integrate into the host genome and to transduce nondividing cells makes them attractive as vehicles for gene delivery. In this study, we assessed the ability of several AAV vectors to transduce airway cells in rabbits by measuring marker gene expression. AAV vectors that transferred either a beta-galactosidase (beta-gal) or a human placental alkaline phosphatase (AP) gene were delivered to one lobe of the rabbit lung by use of a balloon catheter placed under fluoroscopic guidance. We observed vector-encoded beta-gal or AP staining almost exclusively in the epithelial and smooth muscle cells in the bronchus at the region of balloon placement. The overall efficiency of transduction in the balloon-treated bronchial epithelium was low but reached 20% in some areas. The majority of the staining was in ciliated cells but was also observed in basal cells and airway smooth muscle cells. We observed an 80-fold decrease in marker-positive epithelial cells during the 60-day period after vector infusion, whereas the number of marker-positive smooth muscle cells stayed constant. Although treatment with the topoisomerase inhibitor etoposide dramatically enhanced AAV transduction in primary airway epithelial cells in culture, treatment of rabbits did not improve transduction rates in the airway. Vector readministration failed to produce additional transduction events, which correlated with the appearance of neutralizing antibodies. These results indicate that both readministration and immune modulation will be required in the use of AAV vectors for gene therapy to the airway epithelium.

[1]  D. Russell,et al.  Persistent expression of human clotting factor IX from mouse liver after intravenous injection of adeno-associated virus vectors. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[2]  R. Samulski,et al.  Efficient long-term gene transfer into muscle tissue of immunocompetent mice by adeno-associated virus vector , 1996, Journal of virology.

[3]  C. Halbert,et al.  Retroviral vectors efficiently transduce basal and secretory airway epithelial cells in vitro resulting in persistent gene expression in organotypic culture. , 1996, Human gene therapy.

[4]  S. Forman,et al.  Integration of adeno-associated virus vectors in CD34+ human hematopoietic progenitor cells after transduction. , 1996, Blood.

[5]  D. Russell,et al.  Effects of gamma irradiation on the transduction of dividing and nondividing cells in brain and muscle of rats by adeno-associated virus vectors. , 1996, Human gene therapy.

[6]  T. Samulski,et al.  Second-strand synthesis is a rate-limiting step for efficient transduction by recombinant adeno-associated virus vectors , 1996, Journal of virology.

[7]  N. Young,et al.  Adeno-associated virus type 2 binds to a 150-kilodalton cell membrane glycoprotein. , 1996, Virology.

[8]  M. Weitzman,et al.  Transduction with recombinant adeno-associated virus for gene therapy is limited by leading-strand synthesis , 1996, Journal of virology.

[9]  Lynne,et al.  Site-specific integration by adeno-associated virus. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[10]  T. Flotte,et al.  Alveolar stem cell transduction by an adeno-associated viral vector. , 1995, Gene Therapy.

[11]  J. Wilson,et al.  Expression of alpha v beta 5 integrin is necessary for efficient adenovirus-mediated gene transfer in the human airway , 1995, Journal of virology.

[12]  G. Trinchieri,et al.  Recombinant IL–12 prevents formation of blocking IgA antibodies to recombinant adenovirus and allows repeated gene therapy to mouse lung , 1995, Nature Medicine.

[13]  D. Russell,et al.  DNA synthesis and topoisomerase inhibitors increase transduction by adeno-associated virus vectors. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[14]  C. Halbert,et al.  Adeno-associated virus vectors transduce primary cells much less efficiently than immortalized cells , 1995, Journal of virology.

[15]  D. Russell,et al.  DNA-damaging agents greatly increase the transduction of nondividing cells by adeno-associated virus vectors , 1994, Journal of virology.

[16]  James M. Wilson,et al.  Inefficient gene transfer by adenovirus vector to cystic fibrosis airway epithelia of mice and humans , 1994, Nature.

[17]  David W. Russell,et al.  Adeno-associated virus vectors preferentially transduce cells in S phase. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[18]  A. Miller,et al.  Long-term biological response of injured rat carotid artery seeded with smooth muscle cells expressing retrovirally introduced human genes. , 1994, The Journal of clinical investigation.

[19]  Donald W. Pfaff,et al.  Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain , 1994, Nature Genetics.

[20]  T. Flotte,et al.  Stable in vivo expression of the cystic fibrosis transmembrane conductance regulator with an adeno-associated virus vector. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Joseph Zabner,et al.  Adenovirus-mediated gene transfer transiently corrects the chloride transport defect in nasal epithelia of patients with cystic fibrosis , 1993, Cell.

[22]  A. Miller,et al.  Non–invasive liposome–mediated gene delivery can correct the ion transport defect in cystic fibrosis mutant mice , 1993 .

[23]  C. Higgins,et al.  Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy , 1993, Nature.

[24]  T. Flotte,et al.  Gene expression from adeno-associated virus vectors in airway epithelial cells. , 1992, American journal of respiratory cell and molecular biology.

[25]  M. Perricaudet,et al.  In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium , 1992, Cell.

[26]  N. Muzyczka.  Use of adeno-associated virus as a general transduction vector for mammalian cells. , 1992, Current topics in microbiology and immunology.

[27]  J. Wilson,et al.  Reconstitution of tracheal grafts with a genetically modified epithelium. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[28]  R. Craig,et al.  A cystic fibrosis bronchial epithelial cell line: immortalization by adeno-12-SV40 infection. , 1991, American journal of respiratory cell and molecular biology.

[29]  Mitchell L. Drumm,et al.  Correction of the cystic fibrosis defect in vitro by retrovirus-mediated gene transfer , 1990, Cell.

[30]  A. Fire,et al.  A modular set of lacZ fusion vectors for studying gene expression in Caenorhabditis elegans. , 1990, Gene.

[31]  A. Miller,et al.  Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection , 1990, Molecular and cellular biology.

[32]  Samulski,et al.  Helper-free stocks of recombinant adeno-associated viruses: normal integration does not require viral gene expression , 1989, Journal of virology.

[33]  J. Lebkowski,et al.  Adeno-associated virus: a vector system for efficient introduction and integration of DNA into a variety of mammalian cell types , 1988, Molecular and cellular biology.

[34]  P. Collis,et al.  Adeno-associated virus general transduction vectors: analysis of proviral structures , 1988, Journal of virology.

[35]  B. Cullen,et al.  Expression of active, membrane-bound human placental alkaline phosphatase by transfected simian cells. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[36]  Y. Hamajima,et al.  [Toxicity studies of VP 16-213 (I)--Acute toxicity in mice, rats and rabbits]. , 1986, The Journal of toxicological sciences.

[37]  C. Astell,et al.  An Escherichia coli recBCsbcBrecF host permits the deletion-resistant propagation of plasmid clones containing the 5'-terminal palindrome of minute virus of mice. , 1985, Gene.

[38]  N. Muzyczka,et al.  Use of adeno-associated virus as a mammalian DNA cloning vector: transduction of neomycin resistance into mammalian tissue culture cells. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[39]  H. Malke.  T. Maniatis, E. F. Fritsch and J. Sambrook, Molecular Cloning: A Laboratory Manual, X + 545 S., 61 Abb., 28 Tab. Cold Spring Harbor, N. Y. 1982. Cold Spring Harbor Laboratory , 1984 .

[40]  C. Plopper,et al.  Distribution of nonciliated bronchiolar epithelial (Clara) cells in intra- and extrapulmonary airways of the rabbit. , 1983, Experimental lung research.

[41]  R. Samulski,et al.  Cloning of adeno-associated virus into pBR322: rescue of intact virus from the recombinant plasmid in human cells. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[42]  V. Anisimov.  Carcinogenesis and aging. III. The role of age in initiation and promotion of carcinogenesis. , 1982, Experimental pathology.

[43]  S. Kauffman.  Cell proliferation in the mammalian lung. , 1980, International review of experimental pathology.

[44]  F. Graham,et al.  Characteristics of a human cell line transformed by DNA from human adenovirus type 5. , 1977, The Journal of general virology.