Tissue distribution and clonal diversity of the T and B cell repertoire in type 1 diabetes
Howard R. Seay, … , Harlan S. Robins, Todd M. Brusko
Howard R. Seay, … , Harlan S. Robins, Todd M. Brusko
Published December 8, 2016
Citation Information: JCI Insight. 2016;1(20):e88242. https://doi.org/10.1172/jci.insight.88242.
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Research Article Immunology

Tissue distribution and clonal diversity of the T and B cell repertoire in type 1 diabetes

Abstract

The adaptive immune repertoire plays a critical role in type 1 diabetes (T1D) pathogenesis. However, efforts to characterize B cell and T cell receptor (TCR) profiles in T1D subjects have been largely limited to peripheral blood sampling and restricted to known antigens. To address this, we collected pancreatic draining lymph nodes (pLN), “irrelevant” nonpancreatic draining lymph nodes, peripheral blood mononuclear cells (PBMC), and splenocytes from T1D subjects (n = 18) and control donors (n = 9) as well as pancreatic islets from 1 T1D patient; from these tissues, we collected purified CD4+ conventional T cells (Tconv), CD4+ Treg, CD8+ T cells, and B cells. By conducting high-throughput immunosequencing of the TCR β chain (TRB) and B cell receptor (BCR) immunoglobulin heavy chain (IGH) on these samples, we sought to analyze the molecular signature of the lymphocyte populations within these tissues and of T1D. Ultimately, we observed a highly tissue-restricted CD4+ repertoire, while up to 24% of CD8+ clones were shared among tissues. We surveyed our data set for previously described proinsulin- and glutamic acid decarboxylase 65–reactive (GAD65-reactive) receptors, and interestingly, we observed a TRB with homology to a known GAD65-reactive TCR (clone GAD4.13) present in 7 T1D donors (38.9%), representing >25% of all productive TRB within Tconv isolated from the pLN of 1 T1D subject. These data demonstrate diverse receptor signatures at the nucleotide level and enriched autoreactive clones at the amino acid level, supporting the utility of coupling immunosequencing data with knowledge of characterized autoreactive receptors.

Authors

Howard R. Seay, Erik Yusko, Stephanie J. Rothweiler, Lin Zhang, Amanda L. Posgai, Martha Campbell-Thompson, Marissa Vignali, Ryan O. Emerson, John S. Kaddis, Dave Ko, Maki Nakayama, Mia J. Smith, John C. Cambier, Alberto Pugliese, Mark A. Atkinson, Harlan S. Robins, Todd M. Brusko

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Figure 1

Productive clonality for donor samples showing unique T cell receptors (TRB) and B cell receptors (IgH) depicted by heatmap.

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Productive clonality for donor samples showing unique T cell receptors (...
Organ donors are listed along the y axis according to Network for Pancreatic Organ Donors with Diabetes (nPOD) case number and disease status. FACS-isolated cell subsets and tissue source are listed on the x axis. This cohort analysis resulted in an average of 35,822 unique sequences per sample, totaling an average of 46,523 total templates per sample. Dark gray boxes indicate samples that were not available for immunosequencing. Heatmap values depict productive clonality ranging from 0 to 0.6, with red to blue coloring centered at 0.1 (indicated by a black line). Productive clonality is a normalized score based on diversity and sample entropy, with higher values (blue) representing enriched oligoclones (samples with fewer predominant rearrangements). Conversely, clonality values approaching 0 (red) represent samples with highly diverse repertoire. For nPOD 6323, intraislet CD4+ and CD8+ T cells were also available for immunosequencing. †nPOD 6274 classification as T2D is based on a prior diagnosis, despite subsequent resumption of metabolic normalcy following gastric bypass surgery. pLN, pancreatic draining lymph node; iLN, “irrelevant” mesenteric and/or inguinal lymph node; PBMC, peripheral blood mononuclear cells; Tconv, conventional T cells; T1D, type 1 diabetes; T2D, type 2 diabetes; FDB, control, other/Flatbush diabetes; AAb+, autoantibody positive without diabetes.