Cargando…

A mutagenesis study of autoantigen optimization for potential T1D vaccine design

A previously reported autoreactive antigen, termed the X-idiotype, isolated from a unique cell population in Type 1 diabetes (T1D) patients, was found to stimulate their CD4+ T cells. This antigen was previously determined to bind more favorably than insulin and its mimic (insulin superagonist) to H...

Descripción completa

Detalles Bibliográficos
Autores principales: Song, Yi, Bell, David R., Ahmed, Rizwan, Chan, Kevin C., Lee, Sangyun, Hamad, Abdel Rahim A., Zhou, Ruhong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10120010/
https://www.ncbi.nlm.nih.gov/pubmed/37040399
http://dx.doi.org/10.1073/pnas.2214430120
_version_ 1785029106334695424
author Song, Yi
Bell, David R.
Ahmed, Rizwan
Chan, Kevin C.
Lee, Sangyun
Hamad, Abdel Rahim A.
Zhou, Ruhong
author_facet Song, Yi
Bell, David R.
Ahmed, Rizwan
Chan, Kevin C.
Lee, Sangyun
Hamad, Abdel Rahim A.
Zhou, Ruhong
author_sort Song, Yi
collection PubMed
description A previously reported autoreactive antigen, termed the X-idiotype, isolated from a unique cell population in Type 1 diabetes (T1D) patients, was found to stimulate their CD4+ T cells. This antigen was previously determined to bind more favorably than insulin and its mimic (insulin superagonist) to HLA-DQ8, supporting its strong role in CD4+ T cell activation. In this work, we probed HLA-X-idiotype-TCR binding and designed enhanced-reactive pHLA-TCR antigens using an in silico mutagenesis approach which we functionally validated by cell proliferation assays and flow cytometry. From a combination of single, double, and swap mutations, we identified antigen-binding sites p4 and p6 as potential mutation sites for HLA binding affinity enhancement. Site p6 is revealed to favor smaller but more hydrophobic residues than the native tyrosine, such as valine (Y6V) and isoleucine (Y6I), indicating a steric mechanism in binding affinity improvement. Meanwhile, site p4 methionine mutation to hydrophobic residues isoleucine (M4I) or leucine (M4L) modestly increases HLA binding affinity. Select p6 mutations to cysteine (Y6C) or isoleucine (Y6I) exhibit favorable TCR binding affinities, while a swap p5-p6 tyrosine–valine double mutant (V5Y_Y6V) and a p6-p7 glutamine–glutamine double mutant (Y6Q_Y7Q) exhibit enhanced HLA binding affinity but weakened TCR affinity. This work holds relevance to potential T1D antigen-based vaccine design and optimization.
format Online
Article
Text
id pubmed-10120010
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher National Academy of Sciences
record_format MEDLINE/PubMed
spelling pubmed-101200102023-04-22 A mutagenesis study of autoantigen optimization for potential T1D vaccine design Song, Yi Bell, David R. Ahmed, Rizwan Chan, Kevin C. Lee, Sangyun Hamad, Abdel Rahim A. Zhou, Ruhong Proc Natl Acad Sci U S A Biological Sciences A previously reported autoreactive antigen, termed the X-idiotype, isolated from a unique cell population in Type 1 diabetes (T1D) patients, was found to stimulate their CD4+ T cells. This antigen was previously determined to bind more favorably than insulin and its mimic (insulin superagonist) to HLA-DQ8, supporting its strong role in CD4+ T cell activation. In this work, we probed HLA-X-idiotype-TCR binding and designed enhanced-reactive pHLA-TCR antigens using an in silico mutagenesis approach which we functionally validated by cell proliferation assays and flow cytometry. From a combination of single, double, and swap mutations, we identified antigen-binding sites p4 and p6 as potential mutation sites for HLA binding affinity enhancement. Site p6 is revealed to favor smaller but more hydrophobic residues than the native tyrosine, such as valine (Y6V) and isoleucine (Y6I), indicating a steric mechanism in binding affinity improvement. Meanwhile, site p4 methionine mutation to hydrophobic residues isoleucine (M4I) or leucine (M4L) modestly increases HLA binding affinity. Select p6 mutations to cysteine (Y6C) or isoleucine (Y6I) exhibit favorable TCR binding affinities, while a swap p5-p6 tyrosine–valine double mutant (V5Y_Y6V) and a p6-p7 glutamine–glutamine double mutant (Y6Q_Y7Q) exhibit enhanced HLA binding affinity but weakened TCR affinity. This work holds relevance to potential T1D antigen-based vaccine design and optimization. National Academy of Sciences 2023-04-11 2023-04-18 /pmc/articles/PMC10120010/ /pubmed/37040399 http://dx.doi.org/10.1073/pnas.2214430120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Song, Yi
Bell, David R.
Ahmed, Rizwan
Chan, Kevin C.
Lee, Sangyun
Hamad, Abdel Rahim A.
Zhou, Ruhong
A mutagenesis study of autoantigen optimization for potential T1D vaccine design
title A mutagenesis study of autoantigen optimization for potential T1D vaccine design
title_full A mutagenesis study of autoantigen optimization for potential T1D vaccine design
title_fullStr A mutagenesis study of autoantigen optimization for potential T1D vaccine design
title_full_unstemmed A mutagenesis study of autoantigen optimization for potential T1D vaccine design
title_short A mutagenesis study of autoantigen optimization for potential T1D vaccine design
title_sort mutagenesis study of autoantigen optimization for potential t1d vaccine design
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10120010/
https://www.ncbi.nlm.nih.gov/pubmed/37040399
http://dx.doi.org/10.1073/pnas.2214430120
work_keys_str_mv AT songyi amutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT belldavidr amutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT ahmedrizwan amutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT chankevinc amutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT leesangyun amutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT hamadabdelrahima amutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT zhouruhong amutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT songyi mutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT belldavidr mutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT ahmedrizwan mutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT chankevinc mutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT leesangyun mutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT hamadabdelrahima mutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign
AT zhouruhong mutagenesisstudyofautoantigenoptimizationforpotentialt1dvaccinedesign