Details

Autor(en) / Beteiligte

• Ruibal, Paula
• Derksen, Ian
• Wolfswinkel, Marjolein
• Voogd, Linda
• Franken, Kees L. M. C.
• El Hebieshy, Angela F.
• Hall, Thorbald
• Schoufour, Tom A. W.
• Wijdeven, Ruud H.
• Ottenhoff, Tom H. M.
• Scheeren, Ferenc A.
• Joosten, Simone A.

Titel

Thermal‐exchange HLA‐E multimers reveal specificity in HLA‐E and NKG2A/CD94 complex interactions

Ist Teil von

• Immunology, 2023-03, Vol.168 (3), p.526-537

Ort / Verlag

England: Wiley Subscription Services, Inc

Erscheinungsjahr

2023

Quelle

Access via Wiley Online Library

Beschreibungen/Notizen

• There is growing interest in HLA‐E‐restricted T‐cell responses as a possible novel, highly conserved, vaccination targets in the context of infectious and malignant diseases. The developing field of HLA multimers for the detection and study of peptide‐specific T cells has allowed the in‐depth study of TCR repertoires and molecular requirements for efficient antigen presentation and T‐cell activation. In this study, we developed a method for efficient peptide thermal exchange on HLA‐E monomers and multimers allowing the high‐throughput production of HLA‐E multimers. We optimized the thermal‐mediated peptide exchange, and flow cytometry staining conditions for the detection of TCR and NKG2A/CD94 receptors, showing that this novel approach can be used for high‐throughput identification and analysis of HLA‐E‐binding peptides which could be involved in T‐cell and NK cell‐mediated immune responses. Importantly, our analysis of NKG2A/CD94 interaction in the presence of modified peptides led to new molecular insights governing the interaction of HLA‐E with this receptor. In particular, our results reveal that interactions of HLA‐E with NKG2A/CD94 and the TCR involve different residues. Altogether, we present a novel HLA‐E multimer technology based on thermal‐mediated peptide exchange allowing us to investigate the molecular requirements for HLA‐E/peptide interaction with its receptors. In this study, we developed a method for efficient peptide thermal exchange on HLA‐E monomers and multimers allowing the high‐throughput production of HLA‐E multimers. We optimized the thermal‐mediated peptide exchange, and flow cytometry staining conditions for the detection of TCR and NKG2A/CD94 receptors, showing that this novel approach can be used for high throughput identification and analysis of HLA‐E‐binding peptides which could be involved in T‐cell and NK cell‐mediated immune responses. Importantly, our analysis of NKG2A/CD94 interaction in the presence of modified peptides led to new molecular insights governing the interaction of HLA‐E with this receptor.