title: A feasibility study of gene gun mediated immunotherapy for renal cell carcinoma.
abstract: OBJECTIVE
  Gene modified autologous tumor cell vaccines have demonstrated a protective and therapeutic effect in murine tumor model systems. The majority of trials to date have used viral methods of gene transfer for vaccine construction. An alternative approach to transfer genes into tumor cells is to use the gene gun, which is a physical method of gene transfection that produces high levels of gene expression without viral agents. We establish the feasibility of generating cytokine secreting autologous renal tumor cell vaccines for use in gene therapy for metastatic renal cell carcinoma.
  
  
  METHODS
  We obtained 1 cm3 tumor tissue from 12 patients undergoing resection of primary or metastatic renal cell carcinoma. The tumor was disaggregated and placed in culture. The phenotype of the primary renal cell lines was established by microscopy and immunohistochemistry. The 1x10(7) lethally irradiated tumor cells were transfected with plasmid deoxyribonucleic acid containing the human (h) granulocyte-macrophage colony-stimulating factor (GM-CSF) gene under control of a cytomegalovirus promoter using the gene gun. The hGM-CSF production was assayed by enzyme-linked immunosorbent assay in the cell culture media 24 hours after transfection.
  
  
  RESULTS
  Of 12 tumor samples 8 grew rapidly to produce a mean of 1.8x10(8) cells after 4 to 5 passages in culture, which was sufficient to produce between 24 and 32 vaccines. Immunocytochemistry confirmed that all cultures were almost exclusively renal tumor cells. Gene gun mediated transfection of lethally irradiated tumor cells resulted in high levels of hGM-CSF production (mean 330 ng./10(6) cells per 24 hours).
  
  
  CONCLUSIONS
  We have demonstrated the feasibility of producing cytokine secreting tumor cell vaccines from primary and metastatic human renal tumors, and plan to use this approach in phase I clinical trials of gene therapy for metastatic renal cell carcinoma.
authors.affiliation: Department of Surgery, H. Lee Moffitt Cancer Center, Tampa, Florida, USA.
authors.forename: J
authors.initials: J
authors.lastname: Seigne
authors.forename: J
authors.initials: J
authors.lastname: Turner
authors.forename: J
authors.initials: J
authors.lastname: Diaz
authors.forename: J
authors.initials: J
authors.lastname: Hackney
authors.forename: J
authors.initials: J
authors.lastname: Pow-Sang
authors.forename: M
authors.initials: M
authors.lastname: Helal
authors.forename: J
authors.initials: J
authors.lastname: Lockhart
authors.forename: H
authors.initials: H
authors.lastname: Yu
mesh_terms: D019470:Biolistics; D002292:Carcinoma, Renal Cell / Q000628:therapy*; D005240:Feasibility Studies; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D007680:Kidney Neoplasms / Q000628:therapy*; D014407:Tumor Cells, Cultured