Abstract

The aim of this grant proposal is to elucidate at a molecular level the interaction between autoantibodies and TPO. Recombinant TPO-specific F(ab) fragments representing the repertoire of TPO antibodies in the sera of patients with autoimmune thyroid disease will be generated. From these F(ab) fragments, combinatorial libraries of IgGl, IgG4, and/or IgG2 with kappa or lambda light chains (or both) will be constructed, depending on the TPO autoantibody profile present in the individual patient's serum. TPO-specific F(ab) fragments will be analyzed for IgG gene family, affinity and specificity to TPO as well as the relative role of heavy versus light chain in TPO binding. Disease-associated antigenic domains and epitopes on TPO will also be determined. Competition studies will be performed using TPO-specific F(ab) fragments and serum TPO antibodies. The linear epitopes recognized by TPO-specific F(ab) fragments will be determined by screening with a TPO cDNA fragment library. Conformational TPO epitopes recognized by both the recombinant F(ab) fragments and TPO antibodies present in patients' sera will be determined by evaluating the interaction with a series of TPO/myeloperoxidase (MPO) chimeras to be constructed. The relationship between domains and epitopes on TPO recognized by autoantibodies, and manifestations of autoimmune thyroid disease will be evaluated.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK036182-12
Application #
2016175
Study Section
Endocrinology Study Section (END)
Program Officer
Akolkar, Beena
Project Start
1985-07-01
Project End
1997-11-30
Budget Start
1996-12-01
Budget End
1997-11-30
Support Year
12
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
University of California San Francisco
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143

  Publications

Chen, Chun-Rong; Hamidi, Sepehr; Braley-Mullen, Helen et al. (2010) Antibodies to thyroid peroxidase arise spontaneously with age in NOD.H-2h4 mice and appear after thyroglobulin antibodies. Endocrinology 151:4583-93
McLachlan, Sandra M; Rapoport, Basil (2007) Thyroid peroxidase as an autoantigen. Thyroid 17:939-48
Guo, Jin; McLachlan, Sandra M; Rapoport, Basil (2006) Antibodies focused on the human autoantibody immunodominant region are induced by B lymphocytes that constitutively express thyroid peroxidase diverted to the major histocompatibility complex II pathway. Thyroid 16:343-9
Quaratino, Sonia; Ruf, Jean; Osman, Mohamed et al. (2005) Human autoantibodies modulate the T cell epitope repertoire but fail to unmask a pathogenic cryptic epitope. J Immunol 174:557-63
McLachlan, Sandra M; Nagayama, Yuji; Rapoport, Basil (2005) Insight into Graves' hyperthyroidism from animal models. Endocr Rev 26:800-32
Guo, Jin; McLachlan, Sandra M; Pichurin, Pavel N et al. (2005) Relationship between thyroid peroxidase T cell epitope restriction and antibody recognition of the autoantibody immunodominant region in human leukocyte antigen DR3 transgenic mice. Endocrinology 146:4961-7
Chazenbalk, G D; Pichurin, P N; Guo, J et al. (2005) Interactions between the mannose receptor and thyroid autoantigens. Clin Exp Immunol 139:216-24
McLachlan, Sandra M; Braley-Mullen, Helen; Chen, Chun-Rong et al. (2005) Dissociation between iodide-induced thyroiditis and antibody-mediated hyperthyroidism in NOD.H-2h4 mice. Endocrinology 146:294-300
Guo, Jin; Pichurin, Pavel N; Morris, John C et al. (2004) Naked deoxyribonucleic acid vaccination induces recognition of diverse thyroid peroxidase T cell epitopes. Endocrinology 145:3671-8
Latrofa, Francesco; Phillips, Martin; Rapoport, Basil et al. (2004) Human monoclonal thyroglobulin autoantibodies: epitopes and immunoglobulin genes. J Clin Endocrinol Metab 89:5116-23

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