CD4+ regulatory T cells in gastric cancer mucosa are proliferating and express high levels of IL-10 but little TGF-β

Gastric Cancer. 2017 Jan;20(1):116-125. doi: 10.1007/s10120-015-0591-z. Epub 2016 Jan 18.

Abstract

Background: An increase of regulatory T cells, defined as CD25high- and/or FOXP3+-expressing CD4+ T cells, within tumors has been reported in several studies. Tregs promote tumor growth by modulating the antitumor immune response, mainly through inhibition of T-cell-mediated tumor cell killing: this has been suggested to be dependent on IL-10 and/or TGF-β. In stomach cancer, the mechanisms behind the accumulation of Tregs in tumor tissue has not been fully elucidated, and neither has Treg gene expression in situ.

Materials and methods: Stomach tissue from gastric cancer patients undergoing gastric resection was analyzed using flow cytometry and cell sorting, followed by RT-PCR.

Results: We observed that stomach CD4+ FOXP3+ T cells proliferated to a higher degree than CD4+ FOXP3- T cells, which may contribute to Treg accumulation in the mucosa. By analyzing DNA methylation, we demonstrated that both proliferating and nonproliferating FOXP3+ T cells exhibited complete demethylation of the FOXP3 gene, indicating a stable FOXP3 expression in both cell populations. Furthermore, analysis of T-cell populations isolated directly from the tumor and tumor-free mucosa demonstrated that CD4+ CD25high T cells have a higher IL-10/IFN-γ gene expression ratio but express lower levels of TGF-β than CD4+ CD25low/- T cells.

Conclusion: We demonstrate strong proliferation among regulatory CD4+ FOXP3+ CD25high T cells in the gastric cancer mucosa. These local Treg express a suppressive cytokine profile characterized by high IL-10 and low TGF-β and IFN-γ production.

Keywords: Gastric cancer; Helicobacter pylori; IL-10; Proliferation; Regulatory T cell.

MeSH terms

  • Aged
  • Aged, 80 and over
  • Apoptosis
  • Biomarkers, Tumor / genetics
  • CD4-Positive T-Lymphocytes / immunology*
  • Cell Proliferation*
  • DNA Methylation
  • Female
  • Flow Cytometry
  • Follow-Up Studies
  • Forkhead Transcription Factors / metabolism
  • Gastric Mucosa / immunology*
  • Humans
  • Interleukin-10 / genetics*
  • Male
  • Middle Aged
  • Neoplasm Staging
  • Prognosis
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stomach Neoplasms / blood
  • Stomach Neoplasms / genetics
  • Stomach Neoplasms / immunology*
  • Stomach Neoplasms / pathology*
  • T-Lymphocytes, Regulatory / immunology*
  • Transforming Growth Factor beta / genetics*
  • Tumor Cells, Cultured

Substances

  • Biomarkers, Tumor
  • FOXP3 protein, human
  • Forkhead Transcription Factors
  • IL10 protein, human
  • RNA, Messenger
  • Transforming Growth Factor beta
  • Interleukin-10