This proposal examines type A and B complexes of peptide-MHC class II molecules and their corresponding unique T cells. Type A and B pMHCs represent different conformational isomers, generated in different intracellular compartments of the antigen presenting cell (APC): type B pMHC are generated in endocytic vesicles different from the pMHC assembly - or MIIC - compartment. In the case of lysozyme (HEL), the explanation centers on the editing role of H-2DM. The proposal contains two major goals described in overlapping sections. Experiments using type B T cell receptor (TCR) transgenic mice indicate that in vivo, the T cells were activated by HEL administered with inflammatory agents or adjuvants. In vivo and ex vivo experiments will follow the paths that the protein HEL takes in order to generate the type B epitopes, i.e. peptide release, release via exosomes, change in intracellular traffic, and presentation by way of apoptotic material. In vivo experiments include intra-vital microscopy in order to trace the dynamics of antigen presentation of type B (and A) T cells: the type B T cell, because of its uniqueness in being presented in a non-DM site, serves as an indicator of how antigen presentation and T cell stimulation may develop and spread beyond the first cell that takes up the antigen. We propose examining the potential of a type B T cell to induce pathology by using the TCR transgenic mouse genetically crossed to mouse strains that express HEL. Type B T cells directed to an autologous protein evade negative selection: a scenario is envisioned whereby peptides from an autologous tissue protein are released and activate the dormant type B T cells and cause pathology. Strains expressing HEL expressed on beta cells will allow us to examine the pathological potential of the type B T cells, and the conditions that may lead to diabetes development. Finally, the possibilities are considered that type B T cells could have other functional expressions depending on the continuous and systemic release of peptides from HEL. We will examine the MLA11.2 TCR transgenic mice when bred into strains in which HEL is expressed systemically, either in the circulation or in the APC network. Importantly, abundant type B T cells to insulin were found spontaneously in the NOD mouse. This seminal finding may be part of the explanation for the development of diabetic autoimmunity. T cells will be examined for the nature of recognition of insulin epitopes. We will examine presentation of insulin at both a biochemical and functional level, searching for the natural peptides bound to I-Ag7 molecules, as well as peptides in the beta cell granule that may explain the development of type B pMHC complex. We will examine the interactions of insulin and insulin peptides with H2-DM and I-Ag7. The biological role of the type B T cells to insulin peptides will be examined under various situations, focusing in particular on their diabetogenic potential. PROJECT NARRATIVE: This investigation examines the function of a set of T cells that participate in autoimmune reactions. It examines how these T cells are activated. It contains a section examining these T cells in diabetic autoimmunity, i.e. type 1 diabetes mellitus.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI024742-25
Application #
8287551
Study Section
Cellular and Molecular Immunology - A Study Section (CMIA)
Program Officer
Gondre-Lewis, Timothy A
Project Start
1987-05-01
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2014-06-30
Support Year
25
Fiscal Year
2012
Total Cost
$488,089
Indirect Cost
$166,978
Name
Washington University
Department
Pathology
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Calderon, Boris; Carrero, Javier A; Unanue, Emil R (2014) The central role of antigen presentation in islets of Langerhans in autoimmune diabetes. Curr Opin Immunol 26:32-40
Nayak, Deepak K; Calderon, Boris; Vomund, Anthony N et al. (2014) ZnT8-reactive T cells are weakly pathogenic in NOD mice but can participate in diabetes under inflammatory conditions. Diabetes 63:3438-48
Mohan, James F; Unanue, Emil R (2013) A novel pathway of presentation by class II-MHC molecules involving peptides or denatured proteins important in autoimmunity. Mol Immunol 55:166-8
Carrero, Javier A; Calderon, Boris; Towfic, Fadi et al. (2013) Defining the transcriptional and cellular landscape of type 1 diabetes in the NOD mouse. PLoS One 8:e59701
Yang, Chiao-Wen; Unanue, Emil R (2013) Neutrophils control the magnitude and spread of the immune response in a thromboxane A2-mediated process. J Exp Med 210:375-87
Mohan, James F; Calderon, Boris; Anderson, Mark S et al. (2013) Pathogenic CD4? T cells recognizing an unstable peptide of insulin are directly recruited into islets bypassing local lymph nodes. J Exp Med 210:2403-14
Calderon, Boris; Unanue, Emil R (2012) Antigen presentation events in autoimmune diabetes. Curr Opin Immunol 24:119-28
Mohan, James F; Petzold, Shirley J; Unanue, Emil R (2011) Register shifting of an insulin peptide-MHC complex allows diabetogenic T cells to escape thymic deletion. J Exp Med 208:2375-83
Strong, Beverly S I; Unanue, Emil R (2011) Presentation of type B peptide-MHC complexes from hen egg white lysozyme by TLR ligands and type I IFNs independent of H2-DM regulation. J Immunol 187:2193-201
Edelson, Brian T; Bradstreet, Tara R; Wumesh, K C et al. (2011) Batf3-dependent CD11b(low/-) peripheral dendritic cells are GM-CSF-independent and are not required for Th cell priming after subcutaneous immunization. PLoS One 6:e25660

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