Cryptic MHC class I-binding peptides are revealed by aminoglycoside-induced stop codon read-through into the 3' UTR.
Aminoglycosides have been proposed as therapies for genetic disorders caused by nonsense mutations, because of their capacity to enhance translational read-through of premature termination codons (PTCs), thereby permitting expression of functional full-length protein. However, a potential consequence of this strategy is the development of an autoimmune response to HLA-presented epitopes encoded downstream of the PTC or other stop codons. Using a recombinant virus-expression system in tissue culture and in mice, we demonstrate that gentamicin can induce expression and MHC class I presentation of a model epitope encoded downstream of a PTC at levels sufficient to activate CD8(+) T cells. The degree of read-through-derived peptide presentation varies with the sequence of the stop codon and +1 nucleotide. Additionally, we applied a mass spectrometry exploration of the HLA class I peptide repertoire of gentamicin-treated cells and identified multiple peptides derived from read-through of conventional stop codons. These results substantiate the possibility of self-reactivity to cryptic epitopes revealed by stop codon read-through therapies and potentially other therapeutic approaches involving compounds that alter translational fidelity.
Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, PA 19107.
Elliot
E
Goodenough
Tara M
TM
Robinson
Matthew B
MB
Zook
Kevin M
KM
Flanigan
John F
JF
Atkins
Michael T
MT
Howard
Laurence C
LC
Eisenlohr
D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D018389:Codon, Nonsense / Q000235:genetics; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000276:immunology; D005839:Gentamicins / Q000494:pharmacology*; D047468:Immunoprecipitation; D008156:Luciferases; D051379:Mice; D009841:Oligonucleotides / Q000235:genetics; D009842:Oligopeptides / Q000235:genetics* / Q000378:metabolism; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects* / Q000235:genetics; D053719:Tandem Mass Spectrometry
D020413:3' Untranslated Regions; D000911:Antibodies, Monoclonal; D018389:Codon, Nonsense; D018984:Epitopes, T-Lymphocyte; D005839:Gentamicins; C408902:MHC binding peptide; D009841:Oligonucleotides; D009842:Oligopeptides; D008156:Luciferases
antigen presentation; autoimmunity; recoding