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Recombinant glutamic acid decarboxylase (representing the single isoform expressed in human islets) detects IDDM-associated 64,000-Mr autoantibodies

Karlsen, Allan E. LU ; Hagopian, William A. ; Petersen, Jacob S. ; Boel, Esper ; Dyrberg, Thomas ; Grubin, Catherine E. ; Michelsen, Birgitte K. ; Madsen, Ole D. and Lernmark, Åke LU orcid (1992) In Diabetes 41(10). p.1355-1359
Abstract

GAD is an autoantigen in IDDM. Molecular cloning and specific antibodies allowed us to demonstrate that only the lower Mr GAD64 isoform is expressed in human islets, in contrast to human brain, rat islets, and rat brain, all of which express both GA064 and GAD67. Expression of the human islet GAD64 isoform in COS-7 and BHK cells resulted in an enzymatically active rGAD64, which is immunoreactive with diabetic sera comparable with that of the islet 64,000-Mr autoantigen. Immunoprecipitation analyses showed that 21/28 (75%) IDDM sera had rGAD64 antibodies compared with only 1/59 (1.7%) of the healthy control sera. In immunoblot analyses, an SMS serum - but only 1/10 randomly selected IDDM sera - recognized the... (More)

GAD is an autoantigen in IDDM. Molecular cloning and specific antibodies allowed us to demonstrate that only the lower Mr GAD64 isoform is expressed in human islets, in contrast to human brain, rat islets, and rat brain, all of which express both GA064 and GAD67. Expression of the human islet GAD64 isoform in COS-7 and BHK cells resulted in an enzymatically active rGAD64, which is immunoreactive with diabetic sera comparable with that of the islet 64,000-Mr autoantigen. Immunoprecipitation analyses showed that 21/28 (75%) IDDM sera had rGAD64 antibodies compared with only 1/59 (1.7%) of the healthy control sera. In immunoblot analyses, an SMS serum - but only 1/10 randomly selected IDDM sera - recognized the blotted rGAD64 without relation to immunoprecipitation titers. In conclusion, only the GAD64 isoform is expressed in human islets, in contrast to rat islets, which also express the GAD67 isoform. The immunological properties of human rGAD64 are comparable with the native 64,000-Mr islet autoantigen, allowing further studies of the immunopathogenesis of IDDM.

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author
; ; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
in
Diabetes
volume
41
issue
10
pages
1355 - 1359
publisher
American Diabetes Association Inc.
external identifiers
  • pmid:1397711
  • scopus:0026661122
ISSN
0012-1797
DOI
10.2337/diab.41.10.1355
language
English
LU publication?
no
id
70cb8a94-ab8c-4872-8479-5186605218d7
date added to LUP
2019-09-11 09:32:41
date last changed
2024-03-13 08:02:41
@article{70cb8a94-ab8c-4872-8479-5186605218d7,
  abstract     = {{<p>GAD is an autoantigen in IDDM. Molecular cloning and specific antibodies allowed us to demonstrate that only the lower M<sub>r</sub> GAD64 isoform is expressed in human islets, in contrast to human brain, rat islets, and rat brain, all of which express both GA064 and GAD67. Expression of the human islet GAD64 isoform in COS-7 and BHK cells resulted in an enzymatically active rGAD64, which is immunoreactive with diabetic sera comparable with that of the islet 64,000-M<sub>r</sub> autoantigen. Immunoprecipitation analyses showed that 21/28 (75%) IDDM sera had rGAD64 antibodies compared with only 1/59 (1.7%) of the healthy control sera. In immunoblot analyses, an SMS serum - but only 1/10 randomly selected IDDM sera - recognized the blotted rGAD64 without relation to immunoprecipitation titers. In conclusion, only the GAD64 isoform is expressed in human islets, in contrast to rat islets, which also express the GAD67 isoform. The immunological properties of human rGAD64 are comparable with the native 64,000-M<sub>r</sub> islet autoantigen, allowing further studies of the immunopathogenesis of IDDM.</p>}},
  author       = {{Karlsen, Allan E. and Hagopian, William A. and Petersen, Jacob S. and Boel, Esper and Dyrberg, Thomas and Grubin, Catherine E. and Michelsen, Birgitte K. and Madsen, Ole D. and Lernmark, Åke}},
  issn         = {{0012-1797}},
  language     = {{eng}},
  month        = {{01}},
  number       = {{10}},
  pages        = {{1355--1359}},
  publisher    = {{American Diabetes Association Inc.}},
  series       = {{Diabetes}},
  title        = {{Recombinant glutamic acid decarboxylase (representing the single isoform expressed in human islets) detects IDDM-associated 64,000-M<sub>r</sub> autoantibodies}},
  url          = {{http://dx.doi.org/10.2337/diab.41.10.1355}},
  doi          = {{10.2337/diab.41.10.1355}},
  volume       = {{41}},
  year         = {{1992}},
}