Kinetics of cytokine gene expression during the in vitro CTL induction against a murine nonimmunogenic tumor

Anticancer Res. 2000 Jan-Feb;20(1A):363-72.

Abstract

Background: The study of the local environment around tumors, including cellular and endogenous cytokine kinetics may improve our understanding of the biological process occurring during the CTL induction.

Materials and methods: The expression kinetics of ten cytokine genes and immune cell subsets were examined by a RT-PCR and immunohistochemistry using a nonimmunogenic murine tumor and its immunogenic variant (A7).

Results: In a mixed lymphocyte-parent tumor cell culture (MLTC) using in vivo A7-immunized spleen cells, where CTL induction was confirmed on day 5 of cultivation, IL-6 gene expression was promptly activated at 12 h, IL-2 and IL-4 gene expression was prominently up-regulated on days 1 and 2, and then IL-10 and TGF-beta gene expression was down-regulated from day 2 onward. In contrast, in a MLTC using non-immunized spleen cells (a CTL non-induction model), no prominent upregulation of the IL-6, IL-2 or IL-4 gene was detected, and expression of the IL-10 and TGF-beta genes increased steadily. Although numerous dendritic cells were observed in the A7 immunized spleen tissue, lower numbers of dendritic cells were evident in the non-immunized spleen tissue.

Conclusions: A cellular environment that includes the presence of dendritic cells and a cytokine environment that involves the timely production of IL-6, IL-2, and IL-4, and subsequent inhibition of the immunosuppressive cytokines IL-10 and TGF-beta, appear to play an important role in induction of CTL in this model.

MeSH terms

  • Animals
  • Cancer Vaccines / immunology
  • Cells, Cultured
  • Cytokines / biosynthesis*
  • Cytokines / genetics
  • Dendritic Cells / immunology
  • Female
  • Fibrosarcoma / immunology*
  • Fibrosarcoma / pathology
  • Gene Expression Regulation / immunology*
  • Granulocyte-Macrophage Colony-Stimulating Factor / biosynthesis
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics
  • Immunization
  • Immunophenotyping
  • Interferon-gamma / genetics
  • Interleukins / biosynthesis
  • Interleukins / genetics
  • Lymphocyte Activation / genetics*
  • Male
  • Methylnitronitrosoguanidine / toxicity
  • Mice
  • Mice, Inbred C3H
  • Neoplasm Transplantation
  • Reverse Transcriptase Polymerase Chain Reaction
  • Spleen / cytology
  • T-Lymphocytes, Cytotoxic / immunology
  • T-Lymphocytes, Cytotoxic / metabolism*
  • Transforming Growth Factor beta / biosynthesis
  • Transforming Growth Factor beta / genetics
  • Tumor Cells, Cultured / drug effects
  • Tumor Cells, Cultured / immunology

Substances

  • Cancer Vaccines
  • Cytokines
  • Interleukins
  • Transforming Growth Factor beta
  • Methylnitronitrosoguanidine
  • Interferon-gamma
  • Granulocyte-Macrophage Colony-Stimulating Factor