Effects of functional antioxidants on the expansion of gamma delta T-cells and their cellular cytotoxicity against bladder cancer cells

BMC Cancer. 2025 Jun 1;25(1):980. doi: 10.1186/s12885-025-14383-7.

Abstract

Purpose: Results of previous studies have demonstrated that T-cell receptor cross-linking rapidly generates reactive oxygen species, which play essential signaling roles within mitochondria for the antigen-specific expansion of T-cells. However, oxidative stress also causes damage to cellular organelles. Thus, modulating ROS metabolism using antioxidants during naïve T-cell activation may promote the expansion and generation of functional T-cells. Notably, urothelial cancer is a sex-specific malignancy with high mortality rates worldwide. The present study aimed to evaluate the effects of various antioxidants on γδ T-cell proliferation, and the associated cytotoxicity against urothelial carcinoma cells (UCs).

Methods: Over a period of cell induction and expansion, peripheral blood mononuclear cells were cultured with or without different antioxidants, including N-acetyl cysteine (NAC), vitamin C and vitamin E. Subsequently, phenotypic characterization of γδ T-cells and their cytolytic effects against UCs were analyzed by flow cytometry and cell viability assays, respectively.

Results and conclusions: The results revealed that NAC partially inhibited T-cell expansion in a dose-dependent manner. In addition, CD3+/Vγ9+ levels and natural killer group 2D receptor expression were mildly reduced following treatment with a high dose of NAC, whereas CD3+/CD56+ levels and CD314 expression in natural killer-like cells were moderately decreased following treatment with vitamin E. Particularly, the direct co-incubation of bladder cancer cells with γδ T-cells supplemented with antioxidants significantly enhanced bladder cancer cytolysis. Collectively, results of the present study revealed that co-administration of functional antioxidants during γδ T-cell expansion may enhance the quality and efficacy of adoptive T-cell therapies for cancer treatment.

Keywords: Adoptive T-cell therapy; Cytolysis; Multi-functional antioxidant; Oxidative stress; Urothelial carcinoma of the bladder; Γδ T-cell.

MeSH terms

  • Acetylcysteine / pharmacology
  • Antioxidants* / pharmacology
  • Ascorbic Acid / pharmacology
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cytotoxicity, Immunologic* / drug effects
  • Humans
  • Intraepithelial Lymphocytes* / drug effects
  • Intraepithelial Lymphocytes* / immunology
  • Lymphocyte Activation / drug effects
  • Oxidative Stress / drug effects
  • Reactive Oxygen Species / metabolism
  • Receptors, Antigen, T-Cell, gamma-delta* / immunology
  • Receptors, Antigen, T-Cell, gamma-delta* / metabolism
  • T-Lymphocytes* / drug effects
  • T-Lymphocytes* / immunology
  • Urinary Bladder Neoplasms* / drug therapy
  • Urinary Bladder Neoplasms* / immunology
  • Urinary Bladder Neoplasms* / metabolism
  • Urinary Bladder Neoplasms* / pathology
  • Vitamin E / pharmacology

Substances

  • Antioxidants
  • Receptors, Antigen, T-Cell, gamma-delta
  • Reactive Oxygen Species
  • Acetylcysteine
  • Ascorbic Acid
  • Vitamin E