title: IL-23 gene therapy for mouse bladder tumour cell lines.
abstract: OBJECTIVE
  • To evaluate the antitumour effects of IL-23 gene transfer into mouse bladder carcinoma (MBT2) cells. • To investigate the mechanisms underlying the subsequent constitutive secrection of IL-23 by the MBT2 cells
  
  
  METHODS
  • An expression vector containing IL-23 gene was introduced into MBT2 cells by liposome-mediated gene transfer, and secretion of IL-23 was confirmed by ELISA. • The in vivo antitumour effect of IL-23-secreting MBT2 cells (MBT2/IL-23) was examined by injecting the cells into syngeneic C3H mice. • A tumour vaccination study using mitomycin C (MMC)-treated IL-23-secreting MBT2 cells was carried out, and the usefulness of in vivo CD25 depletion for an additional vaccine effect was also investigated. • The mechanisms underlying the antitumour effects were investigated by antibody depletion of CD8 or CD4 T cells, or natural killer cells, and cells infiltrating the tumour sites in vivo were assessed using immunohistochemistry.
  
  
  RESULTS
  • Stable transformants transduced with MBT2/IL-23 secreted IL-23 into the culture supernatant. • Genetically engineered IL-23-secreting MBT2 cells were rejected in syngeneic mice. • MBT2/IL-23-vaccinated mice inhibited the tumour growth of parental MBT2 cells injected at a distant site and this vaccine effect was enhanced by combination with in vivo CD25 depletion by an antibody. • The main effector cells for the direct antitumour effect of MBT2/IL-23 were CD8 T cells, which was shown by in vivo depletion and immunohistochemical study.
  
  
  CONCLUSIONS
  • IL-23-secreting MBT2 cells were rejected in syngeneic mice by the activation of CD8 T cells. • MMC-treated MBT2/IL-23 can have a tumour vaccine effect for parental MBT2 cells, and this effect was enhanced by combination with in vivo CD25 depletion.
authors: Department of Urology, Wakayama Medical University, Wakayama, Japan. kurapon@wakayama-med.ac.jp
authors: Tomomi
authors: T
authors: Kuramoto
authors: Reona
authors: R
authors: Fujii
authors: Hiroshi
authors: H
authors: Nagai
authors: Maria Laura
authors: ML
authors: Belladonna
authors: Takayuki
authors: T
authors: Yoshimoto
authors: Yasuo
authors: Y
authors: Kohjimoto
authors: Takeshi
authors: T
authors: Inagaki
authors: Isao
authors: I
authors: Hara
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D019496:Cancer Vaccines / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D006084:Graft Rejection / Q000276:immunology; D053645:Interleukin-2 Receptor alpha Subunit / Q000276:immunology; D053759:Interleukin-23 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008198:Lymph Nodes / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D008809:Mice, Inbred C3H; D009368:Neoplasm Transplantation; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*
chemical_list: D000911:Antibodies, Monoclonal; D019496:Cancer Vaccines; D053645:Interleukin-2 Receptor alpha Subunit; D053759:Interleukin-23