Host responses and persistence of vector genome following intrabronchial administration of an E1-E3- adenovirus gene transfer vector to normal individuals

Ben Gary Harvey, Neil R. Hackett, Scott Ely, Ronald Crystal

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Adenovirus (Ad)-mediated gene transfer to the respiratory epithelium of experimental animals and to nasal and airway epithelium of individuals with cystic fibrosis is followed by transient gene expression. Extensive studies in experimental animals are consistent with the concept that local cellular host anti-vector immune responses account for this short-term expression, and systemic and local [lung epithelial lining fluid (ELF)] anti-Ad neutralizing antibodies are generated following Ad vector administration to the respiratory epithelial surface. To determine if this paradigm holds in normal humans, a first-generation Ad vector (AdGVCD.10, an E1-E3- Ad serotype 5-based vector coding for the Escherichia coli cytosine deaminase gene) was sprayed locally in escalating doses (8 × 108-8 × 1010 particle units (pu), n = 2/group) into the lung airway epithelium of six normal individuals. Serum, ELF, and endobronchial biopsies were obtained at baseline and at various time points following vector administration. In contrast to the observations in experimental animals in which lung administration of first-generation Ad vectors is followed by strong systemic and local host response, bronchial spray administration of the Ad vector to normal humans showed: (1) minimal inflammation in bronchial biopsies, bronchial brushing, and bronchoalveolar lavage fluid; (2) no blood lymphocyte proliferation in five of six individuals in response to in vitro stimulation with Ad antigens; and (3) no significant increase from baseline in blood or lung ELF anti-Ad neutralizing antibodies. Despite this minimal normal human anti-Ad host response, dose-dependent levels of vector DNA in the airway epithelium were transient. Vector DNA in the targeted airway epithelial cells peaked in a dose-dependent fashion at 0.007 to 1.1 copies/cell at day 7 and declined thereafter, reducing to < 10% of peak levels by 2 weeks. These observations demonstrate both the strengths and the limits of using experimental animals to predict human responses to gene transfer vectors. While the transient nature of Ad vector persistence in the airway epithelium is predicted by most experimental animal studies, respiratory epithelial administration of first-generation Ad vectors at doses up to 1010 pu to airway epithelium of healthy individuals elicits minimal to no detectable systemic and mucosal humoral and cellular immune responses, an observation diametrically opposed to the host responses measured in experimental animals. These findings suggest that, while adaptive anti-Ad immune responses likely play some role in the disappearance of the vector DNA following vector administration to the human lung, other mechanisms may also be involved in the response of humans to Ad gene transfer vectors.

Original languageEnglish
Pages (from-to)206-215
Number of pages10
JournalMolecular Therapy
Volume3
Issue number2
DOIs
Publication statusPublished - 1 Jan 2001
Externally publishedYes

Fingerprint

Adenoviridae
Genome
Genes
Lung
Epithelium
Human Adenoviruses
Bronchoalveolar Lavage Fluid
Neutralizing Antibodies
DNA
Cytosine Deaminase
Biopsy
Respiratory Mucosa
Mucosal Immunity
Nasal Mucosa
Humoral Immunity
Cystic Fibrosis
Cellular Immunity
Epithelial Cells
Observation
Lymphocytes

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

Cite this

Host responses and persistence of vector genome following intrabronchial administration of an E1-E3- adenovirus gene transfer vector to normal individuals. / Harvey, Ben Gary; Hackett, Neil R.; Ely, Scott; Crystal, Ronald.

In: Molecular Therapy, Vol. 3, No. 2, 01.01.2001, p. 206-215.

Research output: Contribution to journalArticle

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