Type IA or insulin dependent diabetes (T1D) is caused by autoimmune destruction of insulin producing ? cells in the pancreatic islets by T lymphocytes. The detection of IgG autoantibodies to insulin and other ? cell autoantigens in pre-T1D suggests that T-B lymphocyte interactions are a critical early event in the loss of immune tolerance that leads to T1D. Research in this laboratory is focused on the function of B lymphocytes that recognize the key ? cell autoantigen, insulin. By engineering NOD mice to express an insulin autoantibody as a B cell receptor transgene, we discovered that B lymphocytes make unappreciated contributions to the pathogenesis of T1D as antigen presenting cells. Anti-insulin B lymphocytes were found to process and present pathogenic epitopes from insulin B chain to diabetogenic T cells. In all previous studies insulin epitopes were identified by T cell responses to synthetic peptides and natural insulin epitopes on diabetogenic MHCII (IAg7 in NOD and DQ8 in humans) were not know. By combining the novel resource of our anti-insulin B cells with advanced proteomics, we have overcome the barrier to detection of natural insulin epitopes and have discovered an unexpected quantity and quality of insulin-related peptides eluted from IAg7 on anti-insulin B cells. Features of the insulin ?immunopeptidome? differ strikingly from synthetic peptides used to mimic epitopes. Pathogenic B chain motifs are in low abundance relative to other insulin-related epitopes and they reside on larger peptides that may support more complex interactions. Surprisingly, a large majority of insulin- related residues from IAg7 reside in multiple epitope clusters encompassing the c-peptide of proinsulin. These findings suggest: i) natural B chain epitopes may reside on larger polypeptides and drive more diverse responses than detected with synthetic peptides (e.g. B9-23); ii) features of c-peptide epitopes eluted from IAg7 indicate anti-insulin B cells capture proinsulin at the site of attack in the islets; iii) properties of proinsulin c-peptide that favor MHCII loading govern thymic education of anti-insulin T cells. These hypotheses will be tested in three aims of the proposal. First, we will identify naturally processed B chain epitopes from IAg7 on B cells that drive unique autoaggressive T cells in T1D. Second, the role of insulitis in loading proinsulin- peptides onto IAg7 in anti-insulin B cells will be determined, and quantitative proteomics will be developed to test this potential biomarker. Third, the functional significance of proinsulin peptides eluted from IAg7 for generation of beneficial T cells that leave the thymus will be assessed. Each of these aims is positioned for rapid translation into human T1D by providing more effective reagents for detection and regulation of autoaggressive T cells, developing a biomarker of early islet invasion, and identifying T cells better suited for maintaining immune tolerance.

Public Health Relevance

This project is directly relevant to improving the diagnosis and treatment of human type 1 diabetes. We now demonstrate that anti-insulin B cells provide a novel means to identify the authentic epitopes from insulin and proinsulin that are recognized by diabetes-causing CD4 T cells. Because this study focuses on anti-insulin B cells, it is especially relevant for younger children with T1D in whom insulin autoimmunity dominates and this group is also experiencing the greatest increase in T1D incidence.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI051448-16
Application #
9382430
Study Section
Hypersensitivity, Autoimmune, and Immune-mediated Diseases Study Section (HAI)
Program Officer
Bourcier, Katarzyna
Project Start
2003-05-01
Project End
2022-12-31
Budget Start
2018-01-23
Budget End
2018-12-31
Support Year
16
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Type
DUNS #
079917897
City
Nashville
State
TN
Country
United States
Zip Code
37232
Cho, Sung Hoon; Raybuck, Ariel L; Stengel, Kristy et al. (2016) Germinal centre hypoxia and regulation of antibody qualities by a hypoxia response system. Nature 537:234-238
Wan, Xiaoxiao; Thomas, James W; Unanue, Emil R (2016) Class-switched anti-insulin antibodies originate from unconventional antigen presentation in multiple lymphoid sites. J Exp Med 213:967-78
Packard, Thomas A; Smith, Mia J; Conrad, Francis J et al. (2016) B Cell Receptor Affinity for Insulin Dictates Autoantigen Acquisition and B Cell Functionality in Autoimmune Diabetes. J Clin Med 5:
Bonami, Rachel H; Thomas, James W (2015) Targeting Anti-Insulin B Cell Receptors Improves Receptor Editing in Type 1 Diabetes-Prone Mice. J Immunol 195:4730-41
Williams, Jonathan M; Bonami, Rachel H; Hulbert, Chrys et al. (2015) Reversing Tolerance in Isotype Switch-Competent Anti-Insulin B Lymphocytes. J Immunol 195:853-64
Bonami, Rachel H; Wolfle, William T; Thomas, James W et al. (2014) NFATc2 (NFAT1) assists BCR-mediated anergy in anti-insulin B cells. Mol Immunol 62:321-8
Kendall, Peggy L; Case, James B; Sullivan, Allison M et al. (2013) Tolerant anti-insulin B cells are effective APCs. J Immunol 190:2519-26
Henry-Bonami, Rachel A; Williams, Jonathan M; Rachakonda, Amita B et al. (2013) B lymphocyte ""original sin"" in the bone marrow enhances islet autoreactivity in type 1 diabetes-prone nonobese diabetic mice. J Immunol 190:5992-6003
Cho, Sung Hoon; Raybuck, Ariel; Wei, Mei et al. (2013) B cell-intrinsic and -extrinsic regulation of antibody responses by PARP14, an intracellular (ADP-ribosyl)transferase. J Immunol 191:3169-78
Lee, Keunwook; Heffington, Lindsey; Jellusova, Julia et al. (2013) Requirement for Rictor in homeostasis and function of mature B lymphoid cells. Blood 122:2369-79

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