In situ interleukin-4 gene expression in cancer patients treated with genetically modified tumor vaccine.
Patients with advanced malignancies, participating in our ongoing phase I interleukin-4 (IL-4) gene therapy protocol at the Pittsburgh Cancer Institute, were vaccinated with irradiated autologous tumor cells together with IL-4 gene-transduced irradiated autologous fibroblasts. The level of expression of the IL-4 gene in cultured transduced and selected fibroblasts and in biopsies obtained from vaccination sites was evaluated using quantitative reverse transcription-polymerase chain reaction (RT-PCR). The number of copies of IL-4 mRNA/ng of total cellular RNA was determined in the transduced fibroblasts. Good agreement was observed between IL-4 message expression, as determined by RT-PCR, and IL-4 production, as determined by enzyme-linked immunosorbent assay (ELISA) in the fibroblast supernatants. Tissue biopsies of multiple vaccination sites were obtained from the patients to determine the level of gene expression in situ for IL-4 and Neo-r. The Neo-r gene was used as a marker for transduced fibroblasts. Two weeks after the first vaccination, mRNA for the IL-4 gene was still detectable in all tissue biopsies. The Neo-r gene was also detectable, indicating the presence of transduced fibroblasts in the biopsy. After the second vaccination, expression of the IL-4 and Neo-r genes was generally the highest on day 1 after vaccine administration and was considerably lower but still detectable on day 14 in all biopsies tested. These data indicate that autologous dermal fibroblasts transduced with the IL-4 and Neo-r genes and used as a source of IL-4 in tumor vaccine are able to express the IL-4 gene in vivo.
Suminami
Y
Y
Pittsburgh Cancer Institute, PA 15213-2582, USA.
Elder
E M
EM
Lotze
M T
MT
Whiteside
T L
TL
D001483:Base Sequence; D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics / Q000628:therapy; D002277:Carcinoma / Q000737:chemistry / Q000235:genetics / Q000628:therapy; D002292:Carcinoma, Renal Cell / Q000737:chemistry / Q000235:genetics / Q000628:therapy; D002460:Cell Line; D004352:Drug Resistance, Microbial / Q000235:genetics / Q000276:immunology; D005347:Fibroblasts; D015972:Gene Expression Regulation, Neoplastic / Q000276:immunology*; D015316:Genetic Therapy; D006801:Humans; D015847:Interleukin-4 / Q000235:genetics*; D008545:Melanoma / Q000737:chemistry / Q000235:genetics / Q000628:therapy; D008969:Molecular Sequence Data; D010957:Plasmids; D012333:RNA, Messenger / Q000378:metabolism; D015203:Reproducibility of Results
D012333:RNA, Messenger; D015847:Interleukin-4