Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor

James N. Kochenderfer, Steven A. Feldman, Yangbing Zhao, Hui Xu, Mary A. Black, Richard A. Morgan, Wyndham H. Wilson, Steven A. Rosenberg

Research output: Contribution to journalArticle

Abstract

T cells can be engineered to express the genes of chimeric antigen receptors (CARs) that recognize tumor-associated antigens. We constructed and compared 2 CARs that contained a single chain variable region moiety that recognized CD19. One CAR contained the signaling moiety of the 4-1BB molecule and the other did not. We selected the CAR that did not contain the 4-1BB moiety for further preclinical development. We demonstrated that gammaretroviruses encoding this receptor could transduce human T cells. Anti-CD19-CAR-transduced CD8 and CD4 T cells produced interferon-γ and interleukin-2 specifically in response to CD19 target cells. The transduced T cells specifically killed primary chronic lymphocytic leukemia (CLL) cells. We transduced T cells from CLL patients that had been previously treated with chemotherapy. We induced these T cells to proliferate sufficiently to provide enough cells for clinical adoptive T cell transfer with a protocol consisting of an initial stimulation with an anti-CD3 monoclonal antibody (OKT3) before transduction followed by a second OKT3 stimulation 7 days after transduction. This protocol was successfully adapted for use in CLL patients with high peripheral blood leukemia cell counts by depleting CD19 cells before the initial OKT3 stimulation. In preparation for a clinical trial that will enroll patients with advanced B cell malignancies, we generated a producer cell clone that produces retroviruses encoding the anti-CD19 CAR, and we produced sufficient retroviral supernatant for the proposed clinical trial under good manufacturing practice conditions.

Original languageEnglish (US)
Pages (from-to)689-702
Number of pages14
JournalJournal of Immunotherapy
Volume32
Issue number7
DOIs
StatePublished - Sep 2009
Externally publishedYes

Fingerprint

CD19 Antigens
Antigen Receptors
T-Lymphocytes
Muromonab-CD3
B-Cell Chronic Lymphocytic Leukemia
Gammaretrovirus
T-Cell Prolymphocytic Leukemia
Clinical Trials
Blood Cell Count
Adoptive Transfer
Neoplasm Antigens
Retroviridae
Interferons
Interleukin-2
Leukemia
B-Lymphocytes
Clone Cells
Drug Therapy

Keywords

  • Adoptive T cell therapy
  • CD19
  • Chimeric antigen receptor
  • Gene therapy
  • T cell,gammaretrovirus

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Cancer Research
  • Pharmacology

Cite this

Kochenderfer, J. N., Feldman, S. A., Zhao, Y., Xu, H., Black, M. A., Morgan, R. A., ... Rosenberg, S. A. (2009). Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor. Journal of Immunotherapy, 32(7), 689-702. https://doi.org/10.1097/CJI.0b013e3181ac6138

Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor. / Kochenderfer, James N.; Feldman, Steven A.; Zhao, Yangbing; Xu, Hui; Black, Mary A.; Morgan, Richard A.; Wilson, Wyndham H.; Rosenberg, Steven A.

In: Journal of Immunotherapy, Vol. 32, No. 7, 09.2009, p. 689-702.

Research output: Contribution to journalArticle

Kochenderfer, JN, Feldman, SA, Zhao, Y, Xu, H, Black, MA, Morgan, RA, Wilson, WH & Rosenberg, SA 2009, 'Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor', Journal of Immunotherapy, vol. 32, no. 7, pp. 689-702. https://doi.org/10.1097/CJI.0b013e3181ac6138
Kochenderfer, James N. ; Feldman, Steven A. ; Zhao, Yangbing ; Xu, Hui ; Black, Mary A. ; Morgan, Richard A. ; Wilson, Wyndham H. ; Rosenberg, Steven A. / Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor. In: Journal of Immunotherapy. 2009 ; Vol. 32, No. 7. pp. 689-702.
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