Quantitative V gene–targeted T cell receptor sequencing as a biomarker in type 1 diabetes
Laurie G. Landry, Kristen L. Wells, Amanda M. Anderson, Kristen R. Miller, Kenneth L. Jones, Aaron W. Michels, Maki Nakayama
Laurie G. Landry, Kristen L. Wells, Amanda M. Anderson, Kristen R. Miller, Kenneth L. Jones, Aaron W. Michels, Maki Nakayama
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Research Article Immunology

Quantitative V gene–targeted T cell receptor sequencing as a biomarker in type 1 diabetes

Abstract

Developing biomarkers to quantitatively monitor disease-specific T cell activity is crucial for assessing type 1 diabetes (T1D) progression and evaluating immunotherapies. This study presents an approach using V gene–targeted sequencing to quantify T cell receptor (TCR) clonotypes as biomarkers for pathogenic T cells in T1D. We identified “public” TCR clonotypes shared among multiple nonobese diabetic (NOD) mice and human organ donors, with a subset expressed exclusively by islet-antigen-reactive T cells in those with T1D. Employing V gene–targeted sequencing of only TCRs containing TRAV16/16D allowed quantitative detection of the public islet-antigen-reactive TCR clonotypes in peripheral blood of NOD mice. Frequencies of these public TCR clonotypes distinguished prediabetic NOD mice from those protected from diabetes. In human islets, public TCR clonotypes identical to preproinsulin-specific clones were exclusively found in T1D donors. This quantifiable TCR sequencing approach uncovered public, disease-specific clonotypes in T1D, providing biomarker candidates to monitor pathogenic T cell frequencies in blood for assessing disease activity and therapeutic response.

Authors

Laurie G. Landry, Kristen L. Wells, Amanda M. Anderson, Kristen R. Miller, Kenneth L. Jones, Aaron W. Michels, Maki Nakayama

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Figure 3

Comparison of TCR repertoires between islets, pancreatic lymph nodes, and peripheral blood.

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Comparison of TCR repertoires between islets, pancreatic lymph nodes, an...
TCR α and β clonotypes in the islets, pancreatic lymph nodes (PLNs), and peripheral blood of 3 prediabetic adult NOD mice were determined and analyzed for frequencies and sharing between the tissues. (A) Individual unique clonotypes are aligned in the order of prevalence in the islets (x axis), and frequencies of clonotypes in the islets were plotted in gray (y axis). Clonotypes that were detected in PLNs and blood samples of the same mouse are marked with red and blue bars, respectively. (B) The association between the frequency of unique clonotypes ranked within 10, 100, and 1,000 in the islets as well as all clonotypes detected in the islets and the percentage of those clonotypes in PLNs (red) and blood samples (blue) are shown. Filled and open triangles represent α and β, respectively. A linear mixed model was fit to the percentage of shared unique clonotypes by rank, accounting for correlation within each mouse. An interaction between categorical rank by group (PLNs vs. blood) was included and significant differences between PLNs and blood samples at each rank were assessed using least square means (Supplemental Table 6). (C) Frequencies of public NY8.3 (top panels), sub-public NY8.3 (panels in between), and all extended NY8.3 clonotypes (bottom panels) in the islets, PLNs, and blood samples that were determined by whole TCR sequencing are shown in left panels. Right panels show frequencies of public, sub-public, and extended NY8.3 clonotypes in blood samples that were determined by TRAV16-targeted sequencing. Each color designates each mouse studied (red NOD26, blue NOD28, and white NOD29).