title: Transfer and expression of the human interleukin-4 gene in carcinoma and stromal cell lines derived from lung cancer patients.
abstract: Introduction of the interleukin-4 (IL-4) gene into cells derived from human tumor tissue provides a means for generating a specific tumor vaccine. Such a vaccine could be produced by either transducing tumor-derived stromal cells with the IL-4 vector and coinjecting tumor cells, or by transducing the tumor cells themselves. We have developed a protocol for culturing cells from non-small cell lung tumors and routinely produce tumor cultures from 25% of tumors, and stromal cultures from > 80% of specimens. Several of these cultures were transduced with the incompetent retroviral vector G1NaSvi4.25, which encodes the human IL-4 cDNA and the G418-resistance gene. Infection of cells by viral titers of 2-5 x 10(4) plaque-forming units/ml, and a multiplicity of infection of 0.1:1 to 1:1 yielded transfer efficiencies of 3.3-32.0 transfectants per 10(4) cells in six of eight attempts. Following selection with the neomycin analog G418, IL-4-producing cells were isolated. IL-4 titers ranged from 142 to 593 U/ml/10(6) in a 24-h collection. Successful transfer of the IL-4 gene was demonstrated by polymerase chain reaction amplification of cDNA derived from reverse-transcribed total RNA, by immunohistochemistry, and by enzyme-linked immunosorbent assay. The IL-4-producing cells were shown to stimulate the proliferation of autologous peripheral blood lymphocytes in one individual by 7.5-fold over control and by 4.1-fold over non-IL-4 producing tumor cells.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Pharmacology, University of Pittsburgh School of Medicine, PA 15261.
authors.forename: J D
authors.initials: JD
authors.lastname: Hunt
authors.forename: B A
authors.initials: BA
authors.lastname: Pippin
authors.forename: R J
authors.initials: RJ
authors.lastname: Landreneau
authors.forename: W F
authors.initials: WF
authors.lastname: Jacob
authors.forename: M T
authors.initials: MT
authors.lastname: Lotze
authors.forename: J M
authors.initials: JM
authors.lastname: Siegfried
mesh_terms: D001483:Base Sequence; D002289:Carcinoma, Non-Small-Cell Lung / Q000276:immunology / Q000628:therapy*; D002460:Cell Line; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D015847:Interleukin-4 / Q000096:biosynthesis / Q000235:genetics*; D008175:Lung Neoplasms / Q000276:immunology / Q000378:metabolism / Q000628:therapy*; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D017154:Stromal Cells / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D015847:Interleukin-4