Avoidance of On-Target Off-Tumor Activation Using a Co-stimulation-Only Chimeric Antigen Receptor.
Chimeric antigen receptors (CARs) combine T cell activation with antibody-mediated tumor antigen specificity, bypassing the need for T cell receptor (TCR) ligation. A limitation of CAR technology is on-target off-tumor toxicity caused by target antigen expression on normal cells. Using GD2 as a model cancer antigen, we hypothesized that this could be minimized by using T cells expressing Vγ9Vδ2 TCR, which recognizes transformed cells in a major histocompatibility complex (MHC)-unrestricted manner, in combination with a co-stimulatory CAR that would function independently of the TCR. An anti-GD2 CAR containing a solitary endodomain derived from the NKG2D adaptor DAP10 was expressed in Vγ9Vδ2+T cells. Differential ligation of the CAR and/or TCR using antibody-coated beads showed that pro-inflammatory cytokine response depended on activation of both receptors. Moreover, in killing assays, GD2-expressing neuroblastoma cells that engaged the Vγ9Vδ2 TCR were efficiently lysed, whereas cells that expressed GD2 equivalently but did not engage the Vγ9Vδ2 TCR were untouched. Differentiation between X-on tumor and X-off tumor offers potential for safer immunotherapy and broader target selection.
Institute of Child Health, University College London, London WC1N 1EH, UK.
Jonathan
J
Fisher
Institute of Child Health, University College London, London WC1N 1EH, UK.
Pierre
P
Abramowski
Institute of Child Health, University College London, London WC1N 1EH, UK.
Nisansala Dilrukshi
ND
Wisidagamage Don
Institute of Child Health, University College London, London WC1N 1EH, UK.
Barry
B
Flutter
Institute of Child Health, University College London, London WC1N 1EH, UK.
Anna
A
Capsomidis
Institute of Child Health, University College London, London WC1N 1EH, UK.
Gordon Weng-Kit
GW
Cheung
Institute of Child Health, University College London, London WC1N 1EH, UK.
Kenth
K
Gustafsson
Institute of Child Health, University College London, London WC1N 1EH, UK. Electronic address: j.anderson@ucl.ac.uk.
John
J
Anderson
D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D018106:CD28 Antigens / Q000235:genetics / Q000276:immunology; D017252:CD3 Complex / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D003602:Cytotoxicity, Immunologic; D005732:Gangliosides / Q000737:chemistry* / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D008213:Lymphocyte Activation; D050506:Mutant Chimeric Proteins / Q000235:genetics* / Q000276:immunology; D009474:Neurons / Q000276:immunology / Q000473:pathology; D015202:Protein Engineering / Q000379:methods; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
D000951:Antigens, Neoplasm; D018106:CD28 Antigens; D017252:CD3 Complex; C121704:CD3 antigen, zeta chain; D005732:Gangliosides; D050506:Mutant Chimeric Proteins; D011948:Receptors, Antigen, T-Cell; C019403:ganglioside, GD2
chimeric antigen receptor; toxicity; γδT cell