Retroviral vector-mediated lymphokine gene transfer into human renal cancer cells.
Effective vaccination against cancer, either for prophylaxis or therapy, has been an elusive goal for years. Cytokine gene therapy offers a novel approach to generate immunogenic tumor cell vaccines. To examine the feasibility of cytokine gene transfer into human renal cancer (RC) cells, we introduced the cDNAs for human interleukin-2 (IL-2) or interferon-gamma (IFN-gamma) into various RC cell lines with retroviral vectors. Using the NIH3T3 amplification assay, no replication competent retroviral particles were detectable in cell culture supernatants taken from gene-modified RC cell lines. Efficient expression of both lymphokines was achieved. Depending on the cell line and the vector construct used, lymphokine gene-modified human RC cell lines released 4 to 29 units/10(6) cells of IL-2, or up to 10 units/10(6) cells of IFN-gamma within 48 h. Fluorescence-activated cell sorter analysis of SK-RC-29 cells releasing IFN-gamma showed increased expression of major histocompatibility complex class I antigen, beta 2-microglobulin, and ICAM-1, as well as induction of major histocompatibility complex class II antigen expression [human leukocyte antigen(HLA)-DR, -DP], but no changes in these cell surface markers were observed with SK-RC-29 cells releasing IL-2. Following in vitro gamma-irradiation with 5,000 or 10,000 rad, growth of lymphokine gene-modified RC cells was abrogated, but their capability to release lymphokine and express lymphokine-induced antigenic determinants, such as HLA-DR, was retained. Tumor formation by the human RC cell line SK-RC-29 in BALB/c nude mice was not affected by IFN-gamma secretion, but was inhibited by in vivo release of IL-2 from s.c. injected tumor cells. These studies demonstrate the feasibility of retroviral mediated lymphokine-gene transfer into human RC cells and suggest a means for generating autologous or HLA-matched allogeneic tumor cell vaccines for the treatment of patients with renal cell carcinoma.
Gastl
G
G
Department of Surgery/Urology, New York Hospital-Cornell Medical Center, New York.
Finstad
C L
CL
Guarini
A
A
Bosl
G
G
Gilboa
E
E
Bander
N H
NH
Gansbacher
B
B
D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000382:microbiology / Q000503:physiopathology; D002470:Cell Survival / Q000528:radiation effects; D004247:DNA / Q000235:genetics; D004307:Dose-Response Relationship, Radiation; D005720:Gamma Rays; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D007680:Kidney Neoplasms / Q000235:genetics* / Q000382:microbiology / Q000503:physiopathology; D008222:Lymphokines / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010641:Phenotype; D012190:Retroviridae / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D014779:Virus Replication / Q000502:physiology
D000951:Antigens, Neoplasm; D007376:Interleukin-2; D008222:Lymphokines; D007371:Interferon-gamma; D004247:DNA