Negative Co-stimulation Constrains T Cell Differentiation by Imposing Boundaries on Possible Cell States

Immunity. 2019 Apr 16;50(4):1084-1098.e10. doi: 10.1016/j.immuni.2019.03.004. Epub 2019 Mar 26.


Co-stimulation regulates T cell activation, but it remains unclear whether co-stimulatory pathways also control T cell differentiation. We used mass cytometry to profile T cells generated in the genetic absence of the negative co-stimulatory molecules CTLA-4 and PD-1. Our data indicate that negative co-stimulation constrains the possible cell states that peripheral T cells can acquire. CTLA-4 imposes major boundaries on CD4+ T cell phenotypes, whereas PD-1 subtly limits CD8+ T cell phenotypes. By computationally reconstructing T cell differentiation paths, we identified protein expression changes that underlied the abnormal phenotypic expansion and pinpointed when lineage choice events occurred during differentiation. Similar alterations in T cell phenotypes were observed after anti-CTLA-4 and anti-PD-1 antibody blockade. These findings implicate negative co-stimulation as a key regulator and determinant of T cell differentiation and suggest that checkpoint blockade might work in part by altering the limits of T cell phenotypes.

Publication types

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

MeSH terms

  • Animals
  • CD4-Positive T-Lymphocytes / classification
  • CD4-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / immunology
  • CTLA-4 Antigen / deficiency
  • CTLA-4 Antigen / genetics
  • CTLA-4 Antigen / immunology*
  • Cell Lineage
  • Immunophenotyping
  • Lymph Nodes / cytology
  • Lymphocyte Activation*
  • Lymphopoiesis*
  • Mice, Knockout
  • Programmed Cell Death 1 Receptor / immunology*
  • T-Lymphocyte Subsets / cytology*
  • Thymus Gland / cytology


  • CTLA-4 Antigen
  • Ctla4 protein, mouse
  • Pdcd1 protein, mouse
  • Programmed Cell Death 1 Receptor