Inducing T cell dysfunction by chronic stimulation of CAR-engineered T cells targeting cancer cells in suspension cultures

Mehmet Emrah Selli; Jack H Landmann; Corvin Arveseth; Nathan Singh

doi:10.1016/j.xpro.2022.101954

Inducing T cell dysfunction by chronic stimulation of CAR-engineered T cells targeting cancer cells in suspension cultures

STAR Protoc. 2023 Mar 17;4(1):101954. doi: 10.1016/j.xpro.2022.101954. Epub 2023 Jan 4.

Authors

Mehmet Emrah Selli¹, Jack H Landmann¹, Corvin Arveseth¹, Nathan Singh²

Affiliations

¹ Washington University School of Medicine, Division of Oncology, St. Louis, MO 63105, USA.
² Washington University School of Medicine, Division of Oncology, St. Louis, MO 63105, USA. Electronic address: nathan.singh@wustl.edu.

Abstract

Several pre-clinical models reveal that chronic chimeric antigen receptor (CAR) stimulation drives a dysfunctional state that mimics in vivo failure. In this protocol, we describe steps to induce T cell dysfunction by persistent and long-term stimulation of CAR-engineered T cells using antigen-expressing cancer cells in suspension cultures. We first described a validated method for manufacturing of CAR T cells, followed by a detailed method for chronic stimulation of CAR T cells and a strategy to evaluate these cells during the process of chronic stimulation. For complete details on the use and execution of this protocol, please refer to Singh et al. (2020).¹.

Keywords: Biotechnology and bioengineering; Cancer; Immunology.

Publication types

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

MeSH terms

Immunologic Tests
Immunotherapy, Adoptive / methods
Neoplasms* / therapy
Receptors, Antigen, T-Cell / genetics
Receptors, Chimeric Antigen* / genetics
T-Lymphocytes

Substances

Receptors, Antigen, T-Cell
Receptors, Chimeric Antigen