In vivo generation of human CD 19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome

• DOI: 10.15252/emmm.201809158
• Publication Date: 2018-09-17T09:55:11Z
• Authors: Anett Pfeiffer; Frederic B Thalhe...; Sylvia Hartmann; Annika M Frank; Ruben R Bender; Simon Danisch; Caroline Costa; Winfried S Wels; Ute Modlich; Renata Stripecke; Els Verhoeyen; Christian J Buchholz
• Keywords: Medicine (General); T-Lymphocytes; Antigens, CD19; 610; Graft vs Host Disease; QH426-470; Lymphocyte Depletion; Mice; 03 medical and health sciences; R5-920; Report; Genetics; Animals; Humans; ddc:610; gene delivery; T‐cell targeting; ddc:610; B-Lymphocytes; 0303 health sciences; Receptors, Chimeric Antigen; 600; cytokine release syndrome; Syndrome; 3. Good health; HEK293 Cells; Leukocytes, Mononuclear; Cytokines; humanized mouse

Chimeric antigen receptor (CAR) T cells brought substantial benefit to patients with B-cell malignancies. Notwithstanding, CAR T-cell manufacturing requires complex procedures impeding the broad supply chain. Here, we provide evidence that human CD19-CAR T cells can be generated directly in vivo using the lentiviral vector CD8-LV specifically targeting human CD8+ cells. Administration into mice xenografted with Raji lymphoma cells and human peripheral blood mononuclear cells led to CAR expression solely in CD8+ T cells and efficacious elimination of CD19+ B cells. Further, upon injection of CD8-LV into mice transplanted with human CD34+ cells, induction of CAR T cells and CD19+ B-cell depletion was observed in 7 out of 10 treated animals. Notably, three mice showed elevated levels of human cytokines in plasma. Tissue-invading CAR T cells and complete elimination of the B-lymphocyte-rich zones in spleen were indicative of a cytokine release syndrome. Our data demonstrate the feasibility of in vivo reprogramming of human CD8+ CAR T cells active against CD19+ cells, yet with similar adverse effects currently notorious in the clinical practice.