High-Throughput Sequencing of Islet-Infiltrating Memory CD4+ T Cells Reveals a Similar Pattern of TCR Vβ Usage in Prediabetic and Diabetic NOD Mice

Idania Marrero,David E Hamm,Joanna D Davies,Matthias G Von Herrath

doi:10.1371/journal.pone.0076546

Idania Marrero, David E Hamm + Show 2 more

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https://doi.org/10.1371/journal.pone.0076546

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Abstract

Autoreactive memory CD4+ T cells play a critical role in the development of type 1 diabetes, but it is not yet known how the clonotypic composition and TCRβ repertoire of the memory CD4+ T cell compartment changes during the transition from prediabetes to diabetes. In this study, we used high-throughput sequencing to analyze the TCRβ repertoire of sorted islet-infiltrating memory CD4+CD44high T cells in 10-week-old prediabetic and recently diabetic NOD mice. We show that most clonotypes of islet-infiltrating CD4+CD44high T cells were rare, but high-frequency clonotypes were significantly more common in diabetic than in prediabetic mice. Moreover, although the CD4+CD44high TCRβ repertoires were highly diverse at both stages of disease development, dominant use of TRBV1 (Vβ2), TRBV13-3 (Vβ8.1), and TRBV19 (Vβ6) was evident in both prediabetic and diabetic mice. Our findings strongly suggest that therapeutic targeting of cells specifically expressing the dominant TCRβ might reduce pancreatic infiltration in prediabetic mice and attenuate the progression to diabetes.

Highlights

Type 1 diabetes (T1D) is an autoimmune disease characterized by selective destruction of insulin-producing b cells within pancreatic islets
Our results reveal a high degree of TCRb diversity among islet-infiltrating CD4+CD44high cells of Nonobese diabetic (NOD) mice and demonstrate that TRBV1, 13-3, and 19 gene segments are dominantly used within the memory repertoire in both prediabetic and diabetic mice
On comparing the CDR3b amino acid sequences of our unique clonotypes with published complementarity-determining region 3 (CDR3) sequences amplified from NOD mouse islets, we found that 12 of the CDR3b sequences using the same TCR b-chain variable (TRBV) and TRBJ gene segment have been reported by others (Table 4, Ref [28,29,30])

Summary

Introduction

Type 1 diabetes (T1D) is an autoimmune disease characterized by selective destruction of insulin-producing b cells within pancreatic islets. Nonobese diabetic (NOD) mice spontaneously develop T1D, and the disease shares many characteristics with human T1D [3]. T cells generally begin to infiltrate the islets at ,3–4 weeks of age and continue to accumulate until b-cell destruction is sufficient to induce overt diabetes at about 12 weeks of age or later [4,5]. Studies examining the T cell receptor (TCR) b repertoire of memory CD4+ T cells in human T1D have been limited mostly to the analysis of peripheral T cells with antigen-specific tetramers. Using GAD65-specific tetramers, Laughlin et al demonstrated that memory CD4+ T cell clones in peripheral blood of two T1D patients with recurrent autoimmunity express identical TCR Vb and complementarity-determining region 3 (CDR3) sequences [7]. In NOD mice, Vb, Db, and Jb gene usage of islet-infiltrating antigen-specific CD4+ T cells has suggested significant skewing of

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