Journal Article

Technical Validation and Utility of an HLA Class II Tetramer Assay for Type 1 Diabetes: A Multicenter Study

Mallone, Roberto, James, Eddie A, Arribas-Layton, David, Ettinger, Ruth A, Overbergh, Lut, Kinnunen, Tuure, Buitinga, Mijke, Speake, Cate, Reijonen, Helena, Afonso, Georgia, Kwok, William W, Bissenova, Samal, Vandamme, Céline, Gomez, Rebecca

2024

Overview

Abstract Context Validated assays to measure autoantigen-specific T-cell frequency and phenotypes are needed for assessing the risk of developing diabetes, monitoring disease progression, evaluating responses to treatment, and personalizing antigen-based therapies. Objective Toward this end, we performed a technical validation of a tetramer assay for HLA-DRA-DRB1*04:01, a class II allele that is strongly associated with susceptibility to type 1 diabetes (T1D). Methods HLA-DRA-DRB1*04:01-restricted T cells specific for immunodominant epitopes from islet cell antigens GAD65, IGRP, preproinsulin, and ZnT8, and a reference influenza epitope, were enumerated and phenotyped in a single staining tube with a tetramer assay. Single and multicenter testing was performed, using a clone-spiked specimen and replicate samples from T1D patients, with a target coefficient of variation (CV) less than 30%. The same assay was applied to an exploratory cross-sectional sample set with 24 T1D patients to evaluate the utility of the assay. Results Influenza-specific T-cell measurements had mean CVs of 6% for the clone-spiked specimen and 11% for T1D samples in single-center testing, and 20% and 31%, respectively, for multicenter testing. Islet-specific T-cell measurements in these same samples had mean CVs of 14% and 23% for single-center and 23% and 41% for multicenter testing. The cross-sectional study identified relationships between T-cell frequencies and phenotype and disease duration, sex, and autoantibodies. A large fraction of the islet-specific T cells exhibited a naive phenotype. Conclusion Our results demonstrate that the assay is reproducible and useful to characterize islet-specific T cells and identify correlations between T-cell measures and clinical traits.

• Publisher: Oxford University Press
• Subject: Antibodies / Antigenic determinants / Autoantibodies / Autoimmunity / Cross-Sectional Studies / Development and progression / Diabetes / Diabetes mellitus (insulin dependent) / Diabetes Mellitus, Type 1 - diagnosis / Diabetes therapy / Diagnosis / Drb1 protein / Epitopes / Ethylenediaminetetraacetic acid / Histocompatibility antigen HLA / Histocompatibility antigens / HLA histocompatibility antigens / HLA-DR alpha-Chains / Humans / Immunodominance / Influenza / Influenza, Human / Lymphocytes / Lymphocytes T / Patients / Phenotypes / Preproinsulin / T cells / T-Lymphocytes / Type 1 diabetes / Viral antibodies