Selective expansion of alveolar macrophages in vivo by adenovirus-mediated transfer of the murine granulocyte-macrophage colony-stimulating factor cDNA. Academic Article uri icon

Overview

MeSH

  • Animals
  • Bronchoalveolar Lavage Fluid
  • DNA, Complementary
  • Female
  • Flow Cytometry
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Luminescent Proteins
  • Lung
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Fluorescence
  • Recombinant Proteins

MeSH Major

  • Adenoviridae
  • Cell Division
  • Gene Expression
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Macrophages, Alveolar
  • Transfection

abstract

  • Based on the hypothesis that genetic modification of freshly isolated alveolar macrophages (AM) with the granulocyte-macrophage colony-stimulating factor (GM-CSF) cDNA would induce AM to proliferate, this study focuses on the ability of adenoviral (Ad) vectors to transfer and efficiently express the murine (m) GM-CSF cDNA in murine AM with consequent expansion in the number of AM in vitro and in vivo. To demonstrate that an Ad vector can effectively transfer and express genes in AM, murine AM recovered by bronchoalveolar lavage from the lung of Balb/c mice were infected with an Ad vector coding for green fluorescent protein (GFP) in vitro and expressed GFP in a dose-dependent fashion. Infection of AM with an Ad vector containing an expression cassette coding for mGM-CSF led to GM-CSF expression and to AM proliferation in vitro. When AM infected with AdGFP were returned to the respiratory tract of syngeneic recipient mice, GFP-expressing cells could still be recovered by bronchoalveolar lavage 2 weeks later. In vitro infection of AM with AdmGM-CSF and subsequent transplantation of the genetically modified AM to the lungs of syngeneic recipients led to GM-CSF expression in vivo. Strikingly, the AM recovered by lavage 5 weeks after transplantation demonstrated an increased rate of proliferation, and the total number of alveolar macrophages was 1. 9-fold greater than controls. Importantly, the increase in the numbers of AM was selective (ie, other inflammatory cell numbers were unchanged), and there was no modification to the lung architecture. Thus, it is feasible to genetically modify AM with Ad vectors and to use this strategy to modify the behavior of AM in vivo. Based on the importance of AM in the primary defense of the respiratory epithelial surface, this strategy may be useful in enhancing pulmonary defenses in immunodeficiency states.

publication date

  • January 15, 1999

has subject area

  • Adenoviridae
  • Animals
  • Bronchoalveolar Lavage Fluid
  • Cell Division
  • DNA, Complementary
  • Female
  • Flow Cytometry
  • Gene Expression
  • Genetic Vectors
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Green Fluorescent Proteins
  • Luminescent Proteins
  • Lung
  • Macrophages, Alveolar
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Fluorescence
  • Recombinant Proteins
  • Transfection

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed ID

  • 9885228

Additional Document Info

start page

  • 655

end page

  • 666

volume

  • 93

number

  • 2