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

Mol Ther. 2017 Feb 1;25(2):456-464. doi: 10.1016/j.ymthe.2016.12.007. Epub 2017 Jan 18.


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 InsB15-232m/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 InsB15-232m/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 InsB15-232m/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.

Keywords: CD8 T cells; NOD mice; immunotherapy; mRNA; type 1 diabetes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer*
  • Animals
  • CD8-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / metabolism*
  • Cytotoxicity, Immunologic
  • Diabetes Mellitus, Type 1 / genetics*
  • Diabetes Mellitus, Type 1 / immunology*
  • Diabetes Mellitus, Type 1 / prevention & control
  • Diabetes Mellitus, Type 1 / therapy
  • Disease Models, Animal
  • Female
  • Gene Expression
  • Gene Order
  • Genetic Vectors / genetics
  • Immunomodulation*
  • Insulin / immunology
  • Mice
  • Mice, Inbred NOD
  • RNA, Messenger / genetics*
  • Receptor-CD3 Complex, Antigen, T-Cell / genetics
  • Recombinant Fusion Proteins / genetics
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism*
  • Transfection
  • beta 2-Microglobulin / genetics


  • Insulin
  • RNA, Messenger
  • Receptor-CD3 Complex, Antigen, T-Cell
  • Recombinant Fusion Proteins
  • beta 2-Microglobulin