Yeast display of MHC-II enables rapid identification of peptide ligands from protein antigens (RIPPA).

Liu, Rongzeng; Jiang, Wei; Mellins, Elizabeth D

Liu, Rongzeng; Jiang, Wei; Mellins, Elizabeth D.

Afiliación

Liu R; Department of Pediatrics-Human Gene Therapy, Stanford University School of Medicine, Stanford, CA, USA.
Jiang W; Stanford Immunology, Stanford University School of Medicine, Stanford, CA, USA.
Mellins ED; Department of Immunology, Henan University of Science and Technology School of Medicine, Luoyang, China.

Cell Mol Immunol ; 18(8): 1847-1860, 2021 08.

Article en En | MEDLINE | ID: mdl-34117370

RESUMEN

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.

Asunto(s)

Linfocitos T CD4-Positivos/metabolismo; COVID-19/metabolismo; Epítopos de Linfocito T/metabolismo; Antígenos HLA-DQ/metabolismo; Antígeno HLA-DR4/metabolismo; Saccharomyces cerevisiae/metabolismo; Glicoproteína de la Espiga del Coronavirus/metabolismo; Técnicas del Sistema de Dos Híbridos; Linfocitos T CD4-Positivos/inmunología; Linfocitos T CD4-Positivos/virología; COVID-19/genética; COVID-19/inmunología; Epítopos de Linfocito T/genética; Epítopos de Linfocito T/inmunología; Citometría de Flujo; Antígenos HLA-DQ/genética; Antígenos HLA-DQ/inmunología; Antígeno HLA-DR4/genética; Antígeno HLA-DR4/inmunología; Ligandos; Saccharomyces cerevisiae/genética; Saccharomyces cerevisiae/inmunología; Glicoproteína de la Espiga del Coronavirus/genética; Glicoproteína de la Espiga del Coronavirus/inmunología

Palabras clave

MHC ligands; RIPPA; SARS-CoV-2 S; T cell epitopes; yeast display

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Saccharomyces cerevisiae / Linfocitos T CD4-Positivos / Antígenos HLA-DQ / Antígeno HLA-DR4 / Epítopos de Linfocito T / Técnicas del Sistema de Dos Híbridos / Glicoproteína de la Espiga del Coronavirus / COVID-19 Tipo de estudio: Diagnostic_studies Idioma: En Revista: Cell Mol Immunol Asunto de la revista: ALERGIA E IMUNOLOGIA Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: China

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google