Transduction of murine and human tumors using recombinant adenovirus vectors

Ann Surg Oncol. 1997 Jan;4(1):70-9. doi: 10.1007/BF02316813.

Abstract

Background: Most cytokine-based cancer gene therapy clinical trials have used labor-intensive, retrovirus-mediated strategies resulting in unpredictable gene expression. Recombinant AdV vectors were evaluated for easier, more reproducible gene transfer into 12 human melanoma, 2 murine fibrosarcomas, and 8 other tumor cell lines.

Methods: AdV vectors contained a reporter (Escherichia coli beta-galactosidase or firefly luciferase) or cytokine gene (human interleukin-2 [IL-2] or IL-7). Transduction efficiencies and expression levels were assessed by histochemical staining, flow cytometry, polymerase chain reaction, fluorometry, and enzyme-linked immunosorbent assay. Tumorigenicity was determined by subcutaneous injection of cells into syngeneic mice.

Results: All cell lines studied were transduced with AdV. Most cell lines exhibited 100% transduction efficiencies (by flow cytometry) at multiplicities of infection (MOI) epsilon 10. Gene expression correlated linearly with MOI, but a cytopathic effect was observed at MOI > 100 with all vectors. Nanogram gene expression levels were routinely achieved. Irradiation (30 Gy) minimally affected expression levels. Tumorigenicity of AdV-IL-2-transduced fibrosarcoma cells in mice was inversely related to IL-2 production. A majority of mice that rejected their tumor challenge were immune to tumor rechallenge.

Conclusions: E1-deleted AdV vectors may prove useful in generating tumor vaccines ex vivo with high, transient cytokine expression levels.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae*
  • Animals
  • Fibrosarcoma / therapy
  • Gene Expression
  • Gene Transfer Techniques*
  • Genes, Reporter
  • Genetic Therapy / methods
  • Genetic Vectors*
  • Histological Techniques
  • Humans
  • Immunohistochemistry
  • Immunotherapy / methods
  • Interleukin-2 / genetics*
  • Interleukin-2 / therapeutic use
  • Interleukin-7 / genetics*
  • Interleukin-7 / therapeutic use
  • Melanoma / therapy
  • Mice
  • Mice, Inbred C3H
  • Neoplasms, Experimental / therapy
  • Tumor Cells, Cultured

Substances

  • Interleukin-2
  • Interleukin-7