The T210M Substitution in the HLA-a*02:01 gp100 Epitope Strongly Affects Overall Proteasomal Cleavage Site Usage and Antigen Processing
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
MHC class I-restricted epitopes, which carry a tumor-specific mutation resulting in improved MHC binding affinity, are preferred T cell receptor targets in innovative adoptive T cell therapies. However, T cell therapy requires efficient generation of the selected epitope. How such mutations may affect proteasome-mediated antigen processing has so far not been studied. Therefore, we analyzed by in vitro experiments the effect on antigen processing and recognition of a T210M exchange, which previously had been introduced into the melanoma gp100209–217tumor epitope to improve the HLA-A*02:01 binding and its immunogenicity. A quantitative analysis of the main steps of antigen processing shows that the T210M exchange affects proteasomal cleavage site usage within the mutgp100201–230 polypeptide, leading to the generation of an unique set of cleavage products. The T210M substitution qualitatively affects the proteasome-catalyzed generation of spliced and non-spliced peptides predicted to bind HLA-A or -B complexes. The T210M substitution also induces an enhanced production of the mutgp100209–217 epitope and its N-terminally extended peptides. The T210M exchange revealed no effect on ERAP1-mediated N-terminal trimming of the precursor peptides. However, mutant N-terminally extended peptides exhibited significantly increased HLA-A*02:01 binding affinity and elicited CD8+ T cell stimulation in vitro similar to the wtgp100209–217 epitope. Thus, our experiments demonstrate that amino acid exchanges within an epitope can result in the generation of an altered peptide pool with new antigenic peptides and in a wider CD8+ T cell response also towards N-terminally extended versions of the minimal epitope.
Date Issued
2015-10-27
Date Acceptance
2015-10-01
Citation
Journal of Biological Chemistry, 2015, 290 (51), pp.30417-30428
ISSN
1083-351X
Publisher
American Society for Biochemistry and Molecular Biology
Start Page
30417
End Page
30428
Journal / Book Title
Journal of Biological Chemistry
Volume
290
Issue
51
Copyright Statement
This research was originally published in The Journal of Biological Chemistry, 2015, vol 290, 30417-30428. © The American Society for Biochemistry and Molecular Biology.
Sponsor
NC3Rs (National Centre for the Replacement, Refinement and Reduction of Animals in Research)
Grant Number
NC/K001949/1
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
CTL EPITOPE
PEPTIDE HYDROLYSIS
PRESENTED PEPTIDES
IMMUNOPROTEASOMES
RECOGNITION
SEQUENCE
BINDING
CELLS
INDUCTION
DIVERSITY
antigen presentation
antigen processing
mutant
protein degradation
ubiquitin-dependent protease
Amino Acid Substitution
Antigen Presentation
CD8-Positive T-Lymphocytes
Cell Line, Transformed
Cell Line, Tumor
Epitopes, T-Lymphocyte
HLA-A2 Antigen
HLA-B Antigens
Humans
Proteasome Endopeptidase Complex
gp100 Melanoma Antigen
06 Biological Sciences
11 Medical And Health Sciences
03 Chemical Sciences
Publication Status
Published