Intratumoral CD4 + T Cells Mediate Anti-tumor Cytotoxicity in Human Bladder Cancer

Cell. 2020 Jun 25;181(7):1612-1625.e13. doi: 10.1016/j.cell.2020.05.017. Epub 2020 Jun 3.


Responses to anti-PD-1 immunotherapy occur but are infrequent in bladder cancer. The specific T cells that mediate tumor rejection are unknown. T cells from human bladder tumors and non-malignant tissue were assessed with single-cell RNA and paired T cell receptor (TCR) sequencing of 30,604 T cells from 7 patients. We find that the states and repertoires of CD8+ T cells are not distinct in tumors compared with non-malignant tissues. In contrast, single-cell analysis of CD4+ T cells demonstrates several tumor-specific states, including multiple distinct states of regulatory T cells. Surprisingly, we also find multiple cytotoxic CD4+ T cell states that are clonally expanded. These CD4+ T cells can kill autologous tumors in an MHC class II-dependent fashion and are suppressed by regulatory T cells. Further, a gene signature of cytotoxic CD4+ T cells in tumors predicts a clinical response in 244 metastatic bladder cancer patients treated with anti-PD-L1.

Trial registration: NCT02451423.

Keywords: Bladder cancer; PD-1 blockade; anti-PD-L1; checkpoint inhibition; cytotoxic CD4(+) T cells; predictive gene signature; single-cell sequencing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • B7-H1 Antigen / genetics
  • B7-H1 Antigen / immunology
  • Biomarkers, Pharmacological / analysis
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / metabolism*
  • CD8-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / metabolism
  • Gene Expression Regulation, Neoplastic / genetics
  • Genes, MHC Class II
  • Humans
  • Immune Checkpoint Inhibitors / pharmacology
  • Immunotherapy
  • Lymphocytes, Tumor-Infiltrating
  • Programmed Cell Death 1 Receptor / genetics
  • Receptors, Antigen, T-Cell / genetics
  • Single-Cell Analysis / methods
  • T-Lymphocytes, Regulatory
  • Urinary Bladder Neoplasms / genetics*
  • Urinary Bladder Neoplasms / immunology
  • Urinary Bladder Neoplasms / metabolism*


  • B7-H1 Antigen
  • Biomarkers, Pharmacological
  • Immune Checkpoint Inhibitors
  • Programmed Cell Death 1 Receptor
  • Receptors, Antigen, T-Cell

Associated data