Retroviral vector-mediated lymphokine gene transfer into human renal cancer cells

Cancer Res. 1992 Nov 15;52(22):6229-36.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antigens, Neoplasm / analysis
  • Carcinoma, Renal Cell / genetics*
  • Carcinoma, Renal Cell / microbiology
  • Carcinoma, Renal Cell / physiopathology
  • Cell Survival / radiation effects
  • DNA / genetics
  • Dose-Response Relationship, Radiation
  • Gamma Rays
  • Gene Expression / genetics
  • Genetic Vectors / genetics
  • Humans
  • Interferon-gamma / biosynthesis
  • Interferon-gamma / genetics
  • Interferon-gamma / metabolism
  • Interleukin-2 / biosynthesis
  • Interleukin-2 / genetics
  • Interleukin-2 / metabolism
  • Kidney Neoplasms / genetics*
  • Kidney Neoplasms / microbiology
  • Kidney Neoplasms / physiopathology
  • Lymphokines / biosynthesis
  • Lymphokines / genetics*
  • Lymphokines / metabolism
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Phenotype
  • Retroviridae / genetics*
  • Retroviridae / physiology
  • Transfection
  • Transplantation, Heterologous
  • Tumor Cells, Cultured
  • Virus Replication / physiology

Substances

  • Antigens, Neoplasm
  • Interleukin-2
  • Lymphokines
  • Interferon-gamma
  • DNA