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 ?₂ microglobulin (?₂m) operates as a universal signaling component of MHC-I molecules when fused with the CD3-? chain. Linking the H-2K^d-binding insulin B chain peptide insulin B chain, amino acids 15-23 (InsB_15-23) to the N terminus of ?₂m/CD3-?, 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/?₂m/CD3-? genes to a reporter T cell line and purified primary mouse CD8 T cells. The peptide/?₂m/CD3-? products paired with endogenous MHC-I chains and transmitted strong activation signals upon MHC-I cross-linking. The reporter T cell line transfected with InsB_15-23/?₂m/CD3-? mRNA was activated by an InsB_15-23-H-2K^d-specific CD8 T cell hybrid only when the transfected T cells expressed H-2K^d. Primary NOD CD8 T cells expressing either InsB_15-23/?₂m/CD3-? or islet-specific glucose-6-phosphatase catalytic subunit-related protein, amino acids 206-214 (IGRP_206-214)/?₂m/CD3-? killed their respective autoreactive CD8 T cell targets in vitro. Furthermore, transfer of primary CD8 T cells transfected with InsB_15-23/?₂m/CD3-? 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.

SUBMITTER: Fishman S 

PROVIDER: S-EPMC5368593 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature