Adoptive Transfer of mRNA-Transfected T Cells Redirected against Diabetogenic CD8 T Cells Can Prevent Diabetes

• Published in: Molecular therapy 2017-02, Vol.25 (2), p.456-464
• Main Authors: Fishman, Sigal, Lewis, Mark D., Siew, L. Khai, De Leenheer, Evy, Kakabadse, Dimitri, Davies, Joanne, Ziv, Doron, Margalit, Alon, Karin, Nathan, Gross, Gideon, Wong, F. Susan
• Format: Article
• Language: English
• Subjects: Adoptive Transfer; Amino acids; Animals; Antigens; beta 2-Microglobulin - genetics; CD3 antigen; CD8 antigen; CD8 T cells; CD8-Positive T-Lymphocytes - immunology; CD8-Positive T-Lymphocytes - metabolism; Cell activation; Cytotoxicity; Cytotoxicity, Immunologic; Diabetes; Diabetes mellitus; Diabetes mellitus (insulin dependent); Diabetes Mellitus, Type 1 - genetics; Diabetes Mellitus, Type 1 - immunology; Diabetes Mellitus, Type 1 - prevention & control; Diabetes Mellitus, Type 1 - therapy; Disease Models, Animal; Electroporation; Experiments; Female; Gene Expression; Gene Order; Genetic Vectors - genetics; Glucose-6-phosphatase; Histocompatibility antigen H-2; Histology; Immunoglobulins; Immunology; Immunomodulation; immunotherapy; Insulin; Insulin - immunology; Insulitis; Kinases; Lymphocytes; Lymphocytes T; Major histocompatibility complex; Mice; Mice, Inbred NOD; mRNA; NOD mice; Original; Peptides; Phosphatase; Polypeptides; Receptor mechanisms; Receptor-CD3 Complex, Antigen, T-Cell - genetics; Recombinant Fusion Proteins - genetics; RNA, Messenger - genetics; T cell receptors; T-Lymphocytes - immunology; T-Lymphocytes - metabolism; Transfection; type 1 diabetes
• ISSN: 1525-0016; 1525-0024
• DOI: 10.1016/j.ymthe.2016.12.007

Chimeric major histocompatibility complex (MHC) molecules supplemented with T cell receptor (TCR) signaling motifs function as activation receptors and can redirect gene-modified T cells against pathogenic CD8 T cells. We have shown that β2 microglobulin (β2m) operates as a universal signaling component of MHC-I molecules when fused with the CD3-ζ chain. Linking the H-2Kd-binding insulin B chain peptide insulin B chain, amino acids 15–23 (InsB15–23) to the N terminus of β2m/CD3-ζ, redirected polyclonal CD8 T cells against pathogenic CD8 T cells in a peptide-specific manner in the non-obese diabetic (NOD) mouse. Here, we describe mRNA electroporation for delivering peptide/β2m/CD3-ζ genes to a reporter T cell line and purified primary mouse CD8 T cells. The peptide/β2m/CD3-ζ products paired with endogenous MHC-I chains and transmitted strong activation signals upon MHC-I cross-linking. The reporter T cell line transfected with InsB15–23/β2m/CD3-ζ mRNA was activated by an InsB15–23-H-2Kd-specific CD8 T cell hybrid only when the transfected T cells expressed H-2Kd. Primary NOD CD8 T cells expressing either InsB15–23/β2m/CD3-ζ or islet-specific glucose-6-phosphatase catalytic subunit-related protein, amino acids 206–214 (IGRP206–214)/β2m/CD3-ζ killed their respective autoreactive CD8 T cell targets in vitro. Furthermore, transfer of primary CD8 T cells transfected with InsB15–23/β2m/CD3-ζ mRNA significantly reduced insulitis and protected NOD mice from diabetes. Our results demonstrate that mRNA encoding chimeric MHC-I receptors can redirect effector CD8 against diabetogenic CD8 T cells, offering a new approach for the treatment of type 1 diabetes. Fishman et al. report a new mRNA-based strategy for redirecting polyclonal CD8 T cells against autoreactive CD8 T cells that cause type 1 diabetes and potentially other autoimmune diseases. Such redirected CD8 T cells activated and killed insulin-specific CD8 T cells in vitro and protected NOD mice from developing diabetes in vivo.