title: Adoptive Transfer of mRNA-Transfected T Cells Redirected against Diabetogenic CD8 T Cells Can Prevent Diabetes.
abstract: 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 β2microglobulin (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.
authors.affiliation: Laboratory of Immunology, MIGAL - Galilee Research Institute, Kiryat Shmona 11016, Israel; Department of Immunology, Rappaport Family Institute for Research in the Medical Sciences, Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, Haifa 3525433, Israel.
authors.forename: Sigal
authors.initials: S
authors.lastname: Fishman
authors.affiliation: Diabetes Research Group, Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff University, Cardiff CF14 4XN, UK.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Lewis
authors.affiliation: Diabetes Research Group, Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff University, Cardiff CF14 4XN, UK.
authors.forename: L Khai
authors.initials: LK
authors.lastname: Siew
authors.affiliation: Diabetes Research Group, Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff University, Cardiff CF14 4XN, UK.
authors.forename: Evy
authors.initials: E
authors.lastname: De Leenheer
authors.affiliation: Diabetes Research Group, Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff University, Cardiff CF14 4XN, UK.
authors.forename: Dimitri
authors.initials: D
authors.lastname: Kakabadse
authors.affiliation: Diabetes Research Group, Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff University, Cardiff CF14 4XN, UK.
authors.forename: Joanne
authors.initials: J
authors.lastname: Davies
authors.affiliation: Laboratory of Immunology, MIGAL - Galilee Research Institute, Kiryat Shmona 11016, Israel; Department of Biotechnology, Tel-Hai College, Upper Galilee 12210, Israel.
authors.forename: Doron
authors.initials: D
authors.lastname: Ziv
authors.affiliation: Laboratory of Immunology, MIGAL - Galilee Research Institute, Kiryat Shmona 11016, Israel; Department of Biotechnology, Tel-Hai College, Upper Galilee 12210, Israel.
authors.forename: Alon
authors.initials: A
authors.lastname: Margalit
authors.affiliation: Department of Immunology, Rappaport Family Institute for Research in the Medical Sciences, Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, Haifa 3525433, Israel.
authors.forename: Nathan
authors.initials: N
authors.lastname: Karin
authors.affiliation: Laboratory of Immunology, MIGAL - Galilee Research Institute, Kiryat Shmona 11016, Israel; Department of Biotechnology, Tel-Hai College, Upper Galilee 12210, Israel. Electronic address: gidi@migal.org.il.
authors.forename: Gideon
authors.initials: G
authors.lastname: Gross
authors.affiliation: Diabetes Research Group, Division of Infection and Immunity, Cardiff University School of Medicine, Cardiff University, Cardiff CF14 4XN, UK.
authors.forename: F Susan
authors.initials: FS
authors.lastname: Wong
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D003602:Cytotoxicity, Immunologic; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics* / Q000276:immunology* / Q000517:prevention & control / Q000628:therapy; D004195:Disease Models, Animal; D005260:Female; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D056747:Immunomodulation; D007328:Insulin / Q000276:immunology; D051379:Mice; D016688:Mice, Inbred NOD; D012333:RNA, Messenger / Q000235:genetics*; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D014162:Transfection; D001613:beta 2-Microglobulin / Q000235:genetics
chemical_list: D007328:Insulin; D012333:RNA, Messenger; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D001613:beta 2-Microglobulin
keywords: CD8 T cells; NOD mice; immunotherapy; mRNA; type 1 diabetes