Human monoclonal antibodies isolated from type I diabetes patients define multiple epitopes in the protein tyrosine phosphatase-like IA-2 antigen

Kolm-Litty, Verena and Berlo, Suzanne and Bonifacio, Ezio and Bearzatto, Massimo and Engel, Alfred M. and Christie, Michael R. and Ziegler, Annette-G. and Wild, Thomas and Endl, Josef (2000) Human monoclonal antibodies isolated from type I diabetes patients define multiple epitopes in the protein tyrosine phosphatase-like IA-2 antigen. Journal of Immunology, 165 (8). pp. 4676-4684. ISSN 0022-1767

Full content URL: http://www.jimmunol.org/content/165/8/4676.full

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Abstract

Protein tyrosine phosphatase-like IA-2 autoantigen is one of the major targets of humoral autoimmunity in patients with insulin-dependant diabetes mellitus (IDDM). In an effort to define the epitopes recognized by autoantibodies against IA-2, we generated five human mAbs (hAbs) from peripheral B lymphocytes isolated from patients most of whom had been recently diagnosed for IDDM. Determination and fine mapping of the critical regions for autoantibody binding was performed by RIA using mutant and chimeric constructs of IA-2- and IA-2β-regions. Four of the five IgG autoantibodies recognized distinct epitopes within the protein tyrosine phosphatase (PTP)-like domain of IA-2. The minimal region required for binding by three of the PTP-like domain-specific hAbs could be located to aa 777-979. Two of these hAbs cross-reacted with the related IA-2β PTP-like domain (IA-2β aa 741-1033). A further PTP-like domain specific hAb required the entire PTP-like domain (aa 687-979) for binding, but critical amino acids clustered in the N-terminal region 687-777. An additional epitope could be localized within the juxtamembrane domain (aa 603-779). In competition experiments, the epitope recognized by one of the hAbs was shown to be targeted by 10 of 14 anti-IA-2-positive sera. Nucleotide sequence analysis of this hAb revealed that it used a V(H) germline gene (DP-71) preferably expressed in autoantibodies associated with IDDM. The presence of somatic mutations in both heavy and light chain genes and the high affinity or this Ab suggest that the immune response to IA-2 is Ag driven.

Keywords:autoantibody, autoantigen, epitope, Ia 2 antigen, Ia 2beta antigen, Ia antibody, immunoglobulin G, monoclonal antibody, protein tyrosine phosphatase, unclassified drug, adolescent, adult, amino acid sequence, amino terminal sequence, antigen binding, antigen detection, antigen recognition, article, autoimmunity, B lymphocyte, child, clinical article, controlled study, human, human cell, insulin dependent diabetes mellitus, nucleotide sequence, priority journal, radioimmunoassay, Adult, Animal, Antibodies, Monoclonal, Autoantibodies, Autoantigens, Base Sequence, Binding Sites, Antibody, Binding, Competitive, Cell Line, Transformed, Diabetes Mellitus, Insulin-Dependent, Epitope Mapping, Genetic Vectors, Immunoglobulin Variable Region, Membrane Proteins, Mice, Middle Age, Molecular Sequence Data, Organ Specificity, Protein-Tyrosine-Phosphatase, Sequence Analysis, Species Specificity
Subjects:C Biological Sciences > C550 Immunology
Divisions:College of Science > School of Life Sciences
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ID Code:18139
Deposited On:07 Aug 2015 09:04

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