A novel ex vivo isolation and expansion procedure for chimeric antigen receptor engrafted human T cells

PLoS One. 2014 Apr 3;9(4):e93745. doi: 10.1371/journal.pone.0093745. eCollection 2014.

Abstract

Genetically engineered T lymphocytes are a promising option for cancer therapy. Prior to adoptive transfer they have to be expanded in vitro to reach therapeutically sufficient numbers. So far, no universal method exists for selective in vitro expansion of engineered T lymphocytes. In order to overcome this problem and for proof of concept we incorporated a novel unique peptide sequence of ten amino acids as epitope (E-Tag) into the binding domains of two novel chimeric antigen receptors (ECARs) directed against either prostate stem cell antigen (PSCA) for the treatment of prostate cancer (PCa) or CD33 for the treatment of acute myeloide leukemia (AML). The epitope tag then was utilized for expanding ECAR engrafted T cells by triggering the modified T cells via a monoclonal antibody directed against the E-Tag (Emab). Moreover, the E-Tag served as an efficient selection epitope for immunomagnetic isolation of modified T cells to high purity. ECAR engrafted T cells were fully functional and mediated profound anti-tumor effects in the respective models of PCa or AML both in vitro and in vivo. The method can be integrated straightforward into clinical protocols to improve therapeutic efficiency of tumor treatment with CAR modified T lymphocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer
  • Epitopes
  • Humans
  • Lymphocyte Activation / immunology*
  • Receptors, Antigen / genetics
  • T-Lymphocytes / immunology*

Substances

  • Epitopes
  • Receptors, Antigen

Grants and funding

This study was supported by grants of the Medical faculty of the TU Dresden to Marc Cartellieri and seed grants given to Michael Bachmann by the Center for Regenerative Therapies Dresden (CRTD), TU Dresden, Dresden, Germany. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.