Alternative Start Sites Downstream of Non-Sense Mutations Drive Antigen Presentation and Tolerance Induction to C-Terminal Epitopes

J Immunol. 2017 Jun 15;198(12):4581-4587. doi: 10.4049/jimmunol.1601131. Epub 2017 May 12.

Abstract

CTL responses to the transgene product remain an active area of concern for the gene therapy field. A patient's underlying genetic mutation may influence the qualitative nature of these potentially destructive T cell responses. Individuals with a mutation that introduces a premature termination codon (PTC) that prevents synthesis of the full-length peptide are considered more likely to mount a transgene-specific T cell response because of a lack of immune tolerance to C-terminal epitopes as a consequence of absent endogenous Ag presentation. In this article, we demonstrate that a human ornithine transcarbamylase gene containing various PTC-inducing non-sense mutations is able to generate and present epitopes downstream of the termination codon. Generation of these epitopes occurs primarily from alternative translation start sites downstream of the stop codon. Furthermore, we show that expression of these genes from adeno-associated virus vectors in C57BL/6 mice is able to induce peripheral tolerance to epitopes downstream of the PTC. These results suggest that, despite the lack of full-length endogenous protein, patients with PTC-inducing non-sense mutations may still present T cell epitopes downstream of the premature termination site that may render the subject tolerant to wild-type transgene products.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antigen Presentation*
  • CD8-Positive T-Lymphocytes
  • Codon, Nonsense*
  • Codon, Terminator*
  • Epitopes, T-Lymphocyte / immunology*
  • Humans
  • Immune Tolerance*
  • Mice
  • Mice, Inbred C57BL
  • Ornithine Carbamoyltransferase / genetics*
  • Ornithine Carbamoyltransferase / metabolism
  • Peptides / genetics
  • Peptides / immunology
  • T-Lymphocytes, Cytotoxic

Substances

  • Codon, Nonsense
  • Codon, Terminator
  • Epitopes, T-Lymphocyte
  • Peptides
  • Ornithine Carbamoyltransferase