Tissue distribution and clonal diversity of the T and B cell repertoire in type 1 diabetes
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
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
View: Text | PDF
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

×

Figure 8

Lymphocyte repertoire diversity was comparable between donors with type 1 diabetes (T1D) and control donors.

Options: View larger image (or click on image) Download as PowerPoint
Lymphocyte repertoire diversity was comparable between donors with type ...
Receptor repertoire diversity was calculated using the Shannon Diversity Index for CD8+ T cells (CD8), B cells (IGH), CD4+ T conventional cells (Tconv), and Treg isolated from (A) pancreatic draining lymph node (pLN), (B) spleen, and (C) “irrelevant” mesenteric and/or inguinal lymph node (iLN), and diversity scores were compared for T1D (gray) versus control (white) subjects (P = NS, all). Data are presented using box-and-whisker plots, with the points representing outliers defined as > (×1.5 IQR + Q3) or < (Q1-1.5 × IQR). T cell receptor (TCR) clones observed in the Treg (x axis) and Tconv (y axis) subsets within the pLN of representative (D) T1D (nPOD 6285), (E) type 2 diabetes (T2D; nPOD 6273), and (F) control (nPOD 6254) subjects are shown in scatter plots, with each point representing a unique clone and its position along the axes representing the clone’s frequency in either subset. Sequences present in only the Tconv subset are to the left of the vertical dotted line, while those detected only in the Treg subset are below the horizontal dotted line. T cell clones common to both subsets are shown in the top right quadrant above and to the right of the dotted lines. (G) Box-and-whisker plots depict the percentage of TCR clones shared between Tconv and Treg subsets within the pLN (median ± distribution, P = NS, Mann-Whitney U test).