Expression of a Chimeric Antigen Receptor Specific for Donor HLA Class I Enhances the Potency of Human Regulatory T Cells in Preventing Human Skin Transplant Rejection

• DOI: 10.1111/ajt.14185
• Publication Date: 2016-12-28T00:08:53Z
• Authors: D.A. Boardman; C. Philippeos; G.O. Fruhwirth; M.A.A. Ibrahim; R.F. Hannen; D. Cooper; F.M. Marelli-Berg; F.M. Watt; R.I. Lechler; J. Maher; L.A. Smyth; G. Lombardi
• Keywords: Graft Rejection; Immune regulation; Molecular biology; basic (laboratory) research; 610; Basic (laboratory) research/science; T-Lymphocytes, Regulatory; T cell biology; Mice; 03 medical and health sciences; Gene therapy; Immunosuppression/immune modulation; 0302 clinical medicine; HLA-A2 Antigen; molecular biology; Animals; Humans; Cellular biology; science; animal models: murine; Mice, Inbred BALB C; immunosuppression; immune modulation; Animal models: murine; immune regulation; Graft Survival; 600; Skin Transplantation; Allografts; gene therapy; murine [Animal models]; Receptors, Antigen; translational research; cellular biology; alloantigen; Leukocytes, Mononuclear; Heterografts; Translational research/science; Transplantation Tolerance; Alloantigen

Regulatory T cell (Treg) therapy using recipient-derived Tregs expanded ex vivo is currently being investigated clinically by us and others as a means of reducing allograft rejection following organ transplantation. Data from animal models has demonstrated that adoptive transfer of allospecific Tregs offers greater protection from graft rejection compared to polyclonal Tregs. Chimeric antigen receptors (CAR) are clinically translatable synthetic fusion proteins that can redirect the specificity of T cells toward designated antigens. We used CAR technology to redirect human polyclonal Tregs toward donor-MHC class I molecules, which are ubiquitously expressed in allografts. Two novel HLA-A2-specific CARs were engineered: one comprising a CD28-CD3ζ signaling domain (CAR) and one lacking an intracellular signaling domain (ΔCAR). CAR Tregs were specifically activated and significantly more suppressive than polyclonal or ΔCAR Tregs in the presence of HLA-A2, without eliciting cytotoxic activity. Furthermore, CAR and ΔCAR Tregs preferentially transmigrated across HLA-A2-expressing endothelial cell monolayers. In a human skin xenograft transplant model, adoptive transfer of CAR Tregs alleviated the alloimmune-mediated skin injury caused by transferring allogeneic peripheral blood mononuclear cells more effectively than polyclonal Tregs. Our results demonstrated that the use of CAR technology is a clinically applicable refinement of Treg therapy for organ transplantation.