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A Comprehensive HLA-DR4 MHC Class II Tetramer Platform for the Detection and Functional Validation of

Henghui Li1, Jingyao Li1, Ying Wang1

  • 1School of Medicine, Shanghai University, Shanghai 200444, China.

International Journal of Molecular Sciences
|July 15, 2026
PubMed

Post-translational modification (PTM) neoantigens have emerged as key drivers of autoimmune inflammation. However, standardized protocols for MHC Class II tetramer preparation for the detection of such antigen-specific T cells remain limited, hindering the broader application of this important discovery. This study systematically engineered an HLA-DR4 (HLA-DRB1*04:02 and HLA-DRA*01:01) tetramer platform based on carboxyethyl-modified neoantigen ITGA2B peptide (ITG-CE), a PTM associated with autoimmune diseases (AUIDs) such as Ankylosing Spondylitis (AS). The platform provides a major histocompatibility complex (MHC) Class II tetramer associated with the PTM neoantigen and integrates modular protein construct, a controllable PTM peptide exchange strategy, and a specific T cell receptor (TCR) validation model. It can be employed to investigate PTM neoantigen presentation and CD4+ T cell auto-reactivity, providing extensive application value for future research into the mechanisms of PTM-induced AUIDs and immune monitoring.

Keywords:
HLA-DR4T-cell receptor (TCR) recognitionantigen-specific CD4+ T cellsautoimmune diseases (AUIDs)fluorescence polarization (FP)high-throughput screeningmajor histocompatibility complex (MHC) class II tetramerpost-translational modification (PTM) neoantigens

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