A role for multiple chimeric antigen receptor-expressing leukocytes in antigen-specific responses to cancer

// Carmen S.M. Yong 1 , Liza B. John 1 , Christel Devaud 1, 2 , Miles H. Prince 1 , Ricky W. Johnstone 1 , Joseph A. Trapani 1 , Phillip K. Darcy 1, 3, * , Michael H. Kershaw 1, 3, * 1 Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Victoria, Australia 2 Institut de Recherche en Santé Digestive, Université de Toulouse, INPT, INRA, INSERM UMR1220, UPS, France 3 Department of Immunology, Monash University, Prahran Victoria, Australia * These authors contributed equally to this work Correspondence to: Phillip K. Darcy, email: phil.darcy@petermac.org Michael H. Kershaw, email: michael.kershaw@petermac.org Keywords: T cells, macrophages, NK cells, chimeric antigen receptor, transgenic mouse Received: March 16, 2016 Accepted: April 16, 2016 Published: May 03, 2016 ABSTRACT While adoptive immunotherapy using chimeric antigen receptor (CAR)-modified T cells can induce remission of some tumors, the role of other CAR-modified leukocytes is not well characterized. In this study, we characterize the function of leukocytes including natural killer (NK) cells, macrophages and CAR T cells from transgenic mice expressing a CAR under the control of the pan-hematopoietic promoter, vav , and determine the ability of these mice to respond to ERB expressing tumors. We demonstrate the anti-tumor functions of leukocytes, including antigen specific cytotoxicity and cytokine secretion. The adoptive transfer of CAR T cells provided a greater survival advantage in the E0771ERB tumor model than their wildtype (WT) counterparts. In addition, CAR NK cells and CAR T cells also mediated increased survival in the RMAERB tumor model. When challenged with Her2 expressing tumors, F38 mice were shown to mount an effective immune response, resulting in tumor rejection and long-term survival. This was shown to be predominantly dependent on both CD8 + T cells and NK cells. However, macrophages and CD4 + T cells were also shown to contribute to this response. Overall, this study highlights the use of the vav -CAR mouse model as a unique tool to determine the anti-tumor function of various immune subsets, either alone or when acting alongside CAR T cells in adoptive immunotherapy.