Yeast display of MHC-II enables rapid identification of peptide ligands from protein antigens (RIPPA).
Cell Mol Immunol
; 18(8): 1847-1860, 2021 08.
Article
in English
| MEDLINE | ID: covidwho-1387308
ABSTRACT
CD4+ T cells orchestrate adaptive immune responses via binding of antigens to their receptors through specific peptide/MHC-II complexes. To study these responses, it is essential to identify protein-derived MHC-II peptide ligands that constitute epitopes for T cell recognition. However, generating cells expressing single MHC-II alleles and isolating these proteins for use in peptide elution or binding studies is time consuming. Here, we express human MHC alleles (HLA-DR4 and HLA-DQ6) as native, noncovalent αß dimers on yeast cells for direct flow cytometry-based screening of peptide ligands from selected antigens. We demonstrate rapid, accurate identification of DQ6 ligands from pre-pro-hypocretin, a narcolepsy-related immunogenic target. We also identify 20 DR4-binding SARS-CoV-2 spike peptides homologous to SARS-CoV-1 epitopes, and one spike peptide overlapping with the reported SARS-CoV-2 epitope recognized by CD4+ T cells from unexposed individuals carrying DR4 subtypes. Our method is optimized for immediate application upon the emergence of novel pathogens.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Saccharomyces cerevisiae
/
CD4-Positive T-Lymphocytes
/
HLA-DQ Antigens
/
HLA-DR4 Antigen
/
Epitopes, T-Lymphocyte
/
Two-Hybrid System Techniques
/
Spike Glycoprotein, Coronavirus
/
COVID-19
Language:
English
Journal:
Cell Mol Immunol
Journal subject:
Allergy and Immunology
Year:
2021
Document Type:
Article
Affiliation country:
S41423-021-00717-5
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