Document Detail

Comparison of the islet cell antibody pattern of monoclonal glutamic acid decarboxylase antibodies recognizing linear and conformational epitopes.
MedLine Citation:
PMID:  8739307     Owner:  NLM     Status:  MEDLINE    
In order to compare the reactivity of glutamic acid decarboxylase (GAD) antibodies recognizing linear and conformational epitopes as islet cell cytoplasmic antibodies (ICA), monoclonal antibodies were generated. An ELISA displacement test using two biotinylated monoclonals recognizing a linear (M61/7E11) or a conformational GAD65 epitope (M65/6B12) was performed to identify epitope regions recognized by monoclonal GAD antibodies. The GAD binding by monoclonal GAD antibodies was tested by immunofluorescence on fixed and unfixed pancreatic sections of human, rat, and mouse, and by Dot-blot experiments. 16/23 (69.6%) of the monoclonals were specifically reactive with GAD65 and 7/23 (30.4%) were reactive with both GAD isoforms. 8/16 (50%) of monoclonal GAD65 antibodies recognized a linear GAD epitope located at the N-terminus (pattern 1). 5/16 (31.3%) displaced M65/6B12, indicating the recognition of a conformational GAD epitope (pattern 2). Monoclonals belonging to patterns 1 and 2 showed strong ICA binding. 3/16 (18.8%) of monoclonals specific for GAD65 with weak or no immunostaining of pancreatic islets (pattern 3) did not inhibit the binding of both biotinylated antibodies in the displacement test, indicating other epitope specificities. In conclusion, GAD antibodies recognizing both conformational and linear epitopes of the GAD65 molecule are involved in ICA binding with strong reactivity. Furthermore, results obtained with monoclonals of pattern 3 suggest the occurrence of GAD65 epitopes partly inaccessible on cryosections, which may result in an ICA-negative test of GAD65 autoantibody positive sera.
P Augstein; M Schlosser; B Ziegler; J Hahmann; L Mauch; M Ziegler
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Acta histochemica     Volume:  98     ISSN:  0065-1281     ISO Abbreviation:  Acta Histochem.     Publication Date:  1996 Apr 
Date Detail:
Created Date:  1997-01-14     Completed Date:  1997-01-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370320     Medline TA:  Acta Histochem     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  229-41     Citation Subset:  IM    
Department of Immunochemistry, Institute of Diabetes Gerhardt Katsch, Karlsburg, University of Greifswald, Germany.
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MeSH Terms
Antibodies, Monoclonal / immunology*
Autoantibodies / immunology*
Blotting, Western
Cerebellum / enzymology
Enzyme-Linked Immunosorbent Assay
Epitopes / chemistry,  immunology*
Fluorescent Antibody Technique
Glutamate Decarboxylase / chemistry*,  immunology,  metabolism
Islets of Langerhans / enzymology,  immunology*
Mice, Inbred BALB C
Pancreas / cytology,  enzymology
Protein Conformation
Rats, Inbred Strains
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Autoantibodies; 0/Epitopes; 0/islet cell antibody; EC Decarboxylase

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