Construction and Preclinical Evaluation of an Anti-CD19 Chimeric Antigen Receptor

作者：James N. Kochenderfer, Steven A. Feldman, Yangbing Zhao, Hui Xu, Mary A. Black, Richard A. Morgan, Wyndham H. Wilson, Steven A. Rosenberg · 发表于：Journal of Immunotherapy · 年份：2009 · DOI：10.1097/cji.0b013e3181ac6138 · 被引用次数：416 · 研究领域：CAR-T cell therapy research、Chronic Lymphocytic Leukemia Research、T-cell and B-cell Immunology

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-gamma 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 prod...