Development and in vitro characterization of a GM-CSF secreting human ovarian carcinoma tumor vaccine

Abstract

A human ovarian carcinoma cell line (UCI-107) was genetically engineered to secrete the cytokine granulocyte-macrophage colony stimulating factor (GM-CSF), by retroviral medicated gene transduction. This line was transduced with the LXSN retroviral vector containing the human GM-CSF gene and the neomycin resistance selection marker. Numerous GM-CSF secreting clones were randomly isolated and one clone, termed UCI-107M GM-CSF-MPS, extensively characterized. This clone was shown to constitutively secrete high levels of GM-CSF (ie 420–585 pg ml−1 105 cells−1 48 h−1 for over 35 passages and 6 months of study. Like the parental cell line UCI-107, UCI-107M GM-CSF-MPS cells expressed MHC class I and Her2/Neu surface antigens but did not express detectable MHC class II, ICAM-1 or CA-125. No change in the expression of these surface proteins was noted between the parental cells and this GM-CSF secreting clone. The morphology of UCI-107M GM-CSF-MPS did not differ from that of the parental or LXSN vector control cells; however, parental cells had a slightly faster growth rate than the transductants. UCI-107M GM-CSF-MPS was sensitive to gamma irradiation, since as little as 2500 rads killed the cells within 10 days of irradiation. However, even after higher doses of irradiation (ie 10000 rads), GM-CSF secretion continuedin vitrountil about day 8. Interestingly, irradiation induced up-regulation of the surface antigens previously expressed, and they remained up-regulated for as long as the cells remained viable. The potential use of these GM-CSF secreting ovarian carcinoma cells as vaccines for women with advanced ovarian cancer will be discussed.