Generation of interleukin-2-secreting melanoma cell populations from resected metastatic tumors.
This study aimed to determine the feasibility of producing patient-specific, interleukin-2 (IL-2)-secreting tumor cell vaccines for the treatment of metastatic melanoma. Primary tumor cell cultures were established from 26/33 resected metastatic melanoma samples. Recombinant retroviral gene transfer and expression in these cultures was optimized using an amphotropic, defective retrovirus carrying the LacZ gene. All cell cultures were infectable; those that proliferated more rapidly were infected at a higher frequency. Addition of fibroblast growth factor to the culture medium increased the rate of cell proliferation and the efficiency of infection. A single infection with an identical retrovirus carrying a human IL-2 cDNA resulted in the generation of unselected cell populations secreting up to 300 units IL-2/10(6) cells.48 hr. Multiple infections increased the level of IL-2 secretion to 5,000 units/10(6) cells.48 hr. The recombinant viral genome could be detected at approximately single copy in the multiply infected cells; no helper virus was detected. IL-2 secretion from infected cultures was maintained following cryopreservation and x-irradiation. These data demonstrate that heterogeneous tumor cell populations secreting IL-2 can be generated from individual patients to be used as autologous, irradiated cell vaccines.
Patel
P M
PM
Chester Beatty Laboratories, Institute of Cancer Research, London, UK.
Flemming
C L
CL
Fisher
C
C
Porter
C D
CD
Thomas
J M
JM
Gore
M E
ME
Collins
M K
MK
D004273:DNA, Neoplasm / Q000096:biosynthesis; D005240:Feasibility Studies; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism*; D007763:Lac Operon / Q000235:genetics; D008545:Melanoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000556:secondary; D014407:Tumor Cells, Cultured / Q000378:metabolism
D004273:DNA, Neoplasm; D007376:Interleukin-2