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Deep exploration of the TCR CDR3β repertoire specific for viral CD4 T-cell epitopes inside the circulating T-cell repertoire
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Deep exploration of the TCR CDR3β repertoire specific for viral CD4 T-cell epitopes inside the circulating T-cell repertoire
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Journal Article
Deep exploration of the TCR CDR3β repertoire specific for viral CD4 T-cell epitopes inside the circulating T-cell repertoire
2025
Overview
This study provides an in-depth analysis of the diversity of the CD4 TCR CDR3β repertoire specific to influenza A (HA) and Epstein-Barr virus (EBNA) epitopes.
Epitope-specific CD4 T cells from 13 healthy donors were enriched using a short-term culture step, isolated based on activation markers, and sequenced for their TCR CDR3β region using high-throughput sequencing. The frequency of each clonotype was then identified within the complete circulating CD4 T-cell CDR3β repertoire.
For both epitopes, the clonotype distribution was markedly skewed, with a small number of highly expanded clones comprising approximately 60% of the repertoire, alongside numerous low-frequency clonotypes. VJ gene usage and motif preferences differed between the two peptides, highlighting epitope-specific TCR selectivity. The response was predominantly composed of private T-cell clonotypes. The proportion of public clonotypes can increase among donors sharing HLA class II molecules and reveals in HLA-unrelated donors the level of TCR promiscuity.
Overall, our data demonstrate that CD4 T-cell responses to these viral epitopes are polyclonal and highly personalized. The modest overlap of clonotypes between donors, coupled with a long tail of low-frequency clones, suggests that the full diversity of the epitope-specific T-cell repertoire is likely broader than previously estimated.
Publisher
Frontiers Media SA,Frontiers Media S.A
Subject
/ Antigens
/ CD4-Positive T-Lymphocytes - immunology
/ Cloning
/ Complementarity Determining Regions - genetics
/ Complementarity Determining Regions - immunology
/ Epitopes
/ Epitopes, T-Lymphocyte - genetics
/ Epitopes, T-Lymphocyte - immunology
/ Epstein-Barr Virus Nuclear Antigens - immunology
/ Female
/ Hemagglutinin Glycoproteins, Influenza Virus - immunology
/ Herpesvirus 4, Human - immunology
/ High-Throughput Nucleotide Sequencing
/ Histocompatibility antigen HLA
/ Humans
/ Influenza A virus - immunology
/ Male
/ Peptides
/ Receptors, Antigen, T-Cell, alpha-beta - genetics
/ Receptors, Antigen, T-Cell, alpha-beta - immunology
/ TCR
/ viral
