Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1993 Sep;92(3):1394–1399. doi: 10.1172/JCI116714

Prevalence of autoantibodies to the 65- and 67-kD isoforms of glutamate decarboxylase in insulin-dependent diabetes mellitus.

J Seissler 1, J Amann 1, L Mauch 1, H Haubruck 1, S Wolfahrt 1, S Bieg 1, W Richter 1, R Holl 1, E Heinze 1, W Northemann 1, et al.
PMCID: PMC288282  PMID: 8376591

Abstract

We investigated the presence of autoantibodies to baculovirus-expressed human recombinant 65- and 67-kD isoforms of glutamate decarboxylase (GAD65 and GAD67) in insulin-dependent diabetes mellitus (IDDM). In the immunoprecipitation test using [35S]methionine-labeled GADs antibodies to GAD65 were detected in 13/15 (87%) islet cell antibody (ICA)-positive and in 1/35 (2.9%) ICA-negative first-degree relatives of patients with IDDM, in 6/11 (54.5%) ICA-positive nondiabetic schoolchildren, and in 35/50 (70%) patients with newly diagnosed IDDM. GAD67 antibodies were positive only in five (33%) of the ICA-positive relatives (P < 0.05) and in nine (18%) IDDM patients at onset (P < 0.00001). After onset of IDDM antibodies to GAD65 and GAD67 declined but were still positive in 25 and 9.4% of subjects with long-standing IDDM (> 10 yr). In all study groups antibodies to GAD67 were only detected in GAD65 antibody-positive sera. An immunotrapping enzyme activity assay for GAD65 antibodies was positive in 64/75 (85.3%) of sera that were GAD antibody positive in the immunoprecipitation test (r = 0.870, P < 0.0001). In two (2.7%) sera GAD65 antibodies that block GAD enzyme activity were found. Our data suggest that antibodies to GAD65 but not to GAD67 represent sensitive markers for preclinical and overt IDDM. The immunotrapping assay here described represents a valuable technique for specific and sensitive screening for GAD antibodies.

Full text

PDF
1394

Images in this article

1395Image
on p.1395

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atkinson M. A., Maclaren N. K., Scharp D. W., Lacy P. E., Riley W. J. 64,000 Mr autoantibodies as predictors of insulin-dependent diabetes. Lancet. 1990 Jun 9;335(8702):1357–1360. doi: 10.1016/0140-6736(90)91241-2. [DOI] [PubMed] [Google Scholar]
  2. Baekkeskov S., Aanstoot H. J., Christgau S., Reetz A., Solimena M., Cascalho M., Folli F., Richter-Olesen H., De Camilli P., Camilli P. D. Identification of the 64K autoantigen in insulin-dependent diabetes as the GABA-synthesizing enzyme glutamic acid decarboxylase. Nature. 1990 Sep 13;347(6289):151–156. doi: 10.1038/347151a0. [DOI] [PubMed] [Google Scholar]
  3. Baekkeskov S., Landin M., Kristensen J. K., Srikanta S., Bruining G. J., Mandrup-Poulsen T., de Beaufort C., Soeldner J. S., Eisenbarth G., Lindgren F. Antibodies to a 64,000 Mr human islet cell antigen precede the clinical onset of insulin-dependent diabetes. J Clin Invest. 1987 Mar;79(3):926–934. doi: 10.1172/JCI112903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boehm B. O., Manfras B., Seissler J., Schöffling K., Glück M., Holzberger G., Seidl S., Kühnl P., Trucco M., Scherbaum W. A. Epidemiology and immunogenetic background of islet cell antibody--positive nondiabetic schoolchildren. Ulm-Frankfurt population study. Diabetes. 1991 Nov;40(11):1435–1439. doi: 10.2337/diab.40.11.1435. [DOI] [PubMed] [Google Scholar]
  5. Bonifacio E., Bingley P. J., Shattock M., Dean B. M., Dunger D., Gale E. A., Bottazzo G. F. Quantification of islet-cell antibodies and prediction of insulin-dependent diabetes. Lancet. 1990 Jan 20;335(8682):147–149. doi: 10.1016/0140-6736(90)90013-u. [DOI] [PubMed] [Google Scholar]
  6. Bärmeier H., McCulloch D. K., Neifing J. L., Warnock G., Rajotte R. V., Palmer J. P., Lernmark A. Risk for developing type 1 (insulin-dependent) diabetes mellitus and the presence of islet 64K antibodies. Diabetologia. 1991 Oct;34(10):727–733. doi: 10.1007/BF00401518. [DOI] [PubMed] [Google Scholar]
  7. Chang Y. C., Gottlieb D. I. Characterization of the proteins purified with monoclonal antibodies to glutamic acid decarboxylase. J Neurosci. 1988 Jun;8(6):2123–2130. doi: 10.1523/JNEUROSCI.08-06-02123.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Christgau S., Schierbeck H., Aanstoot H. J., Aagaard L., Begley K., Kofod H., Hejnaes K., Baekkeskov S. Pancreatic beta cells express two autoantigenic forms of glutamic acid decarboxylase, a 65-kDa hydrophilic form and a 64-kDa amphiphilic form which can be both membrane-bound and soluble. J Biol Chem. 1991 Nov 5;266(31):21257–21264. [PubMed] [Google Scholar]
  9. Christie M. R., Brown T. J., Cassidy D. Binding of antibodies in sera from Type 1 (insulin-dependent) diabetic patients to glutamate decarboxylase from rat tissues. Evidence for antigenic and non-antigenic forms of the enzyme. Diabetologia. 1992 Apr;35(4):380–384. doi: 10.1007/BF00401206. [DOI] [PubMed] [Google Scholar]
  10. Christie M. R., Daneman D., Champagne P., Delovitch T. L. Persistence of serum antibodies to 64,000-Mr islet cell protein after onset of type I diabetes. Diabetes. 1990 Jun;39(6):653–656. doi: 10.2337/diab.39.6.653. [DOI] [PubMed] [Google Scholar]
  11. Christie M., Landin-Olsson M., Sundkvist G., Dahlquist G., Lernmark A., Baekkeskov S. Antibodies to a Mr-64,000 islet cell protein in Swedish children with newly diagnosed type 1 (insulin-dependent) diabetes. Diabetologia. 1988 Aug;31(8):597–602. doi: 10.1007/BF00264766. [DOI] [PubMed] [Google Scholar]
  12. De Aizpurua H. J., Wilson Y. M., Harrison L. C. Glutamic acid decarboxylase autoantibodies in preclinical insulin-dependent diabetes. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9841–9845. doi: 10.1073/pnas.89.20.9841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. DeAizpurua H. J., Harrison L. C., Cram D. S. An ELISA for antibodies to recombinant glutamic acid decarboxylase in IDDM. Diabetes. 1992 Sep;41(9):1182–1187. doi: 10.2337/diab.41.9.1182. [DOI] [PubMed] [Google Scholar]
  14. Erlander M. G., Tillakaratne N. J., Feldblum S., Patel N., Tobin A. J. Two genes encode distinct glutamate decarboxylases. Neuron. 1991 Jul;7(1):91–100. doi: 10.1016/0896-6273(91)90077-d. [DOI] [PubMed] [Google Scholar]
  15. Gottlieb D. I., Chang Y. C., Schwob J. E. Monoclonal antibodies to glutamic acid decarboxylase. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8808–8812. doi: 10.1073/pnas.83.22.8808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Karlsen A. E., Hagopian W. A., Grubin C. E., Dube S., Disteche C. M., Adler D. A., Bärmeier H., Mathewes S., Grant F. J., Foster D. Cloning and primary structure of a human islet isoform of glutamic acid decarboxylase from chromosome 10. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8337–8341. doi: 10.1073/pnas.88.19.8337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaufman D. L., Erlander M. G., Clare-Salzler M., Atkinson M. A., Maclaren N. K., Tobin A. J. Autoimmunity to two forms of glutamate decarboxylase in insulin-dependent diabetes mellitus. J Clin Invest. 1992 Jan;89(1):283–292. doi: 10.1172/JCI115573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaufman D. L., Houser C. R., Tobin A. J. Two forms of the gamma-aminobutyric acid synthetic enzyme glutamate decarboxylase have distinct intraneuronal distributions and cofactor interactions. J Neurochem. 1991 Feb;56(2):720–723. doi: 10.1111/j.1471-4159.1991.tb08211.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Landin-Olsson M., Palmer J. P., Lernmark A., Blom L., Sundkvist G., Nyström L., Dahlquist G. Predictive value of islet cell and insulin autoantibodies for type 1 (insulin-dependent) diabetes mellitus in a population-based study of newly-diagnosed diabetic and matched control children. Diabetologia. 1992 Nov;35(11):1068–1073. doi: 10.1007/BF02221683. [DOI] [PubMed] [Google Scholar]
  20. Martino G. V., Tappaz M. L., Braghi S., Dozio N., Canal N., Pozza G., Bottazzo G. F., Grimaldi L. M., Bosi E. Autoantibodies to glutamic acid decarboxylase (GAD) detected by an immuno-trapping enzyme activity assay: relation to insulin-dependent diabetes mellitus and islet cell antibodies. J Autoimmun. 1991 Dec;4(6):915–923. doi: 10.1016/0896-8411(91)90054-g. [DOI] [PubMed] [Google Scholar]
  21. Miller L. P., Martin D. L., Mazumder A., Walters J. R. Studies on the regulation of GABA synthesis: substrate-promoted dissociation of pyridoxal-5'-phosphate from GAD. J Neurochem. 1978 Feb;30(2):361–369. doi: 10.1111/j.1471-4159.1978.tb06538.x. [DOI] [PubMed] [Google Scholar]
  22. Richter W., Endl J., Eiermann T. H., Brandt M., Kientsch-Engel R., Thivolet C., Jungfer H., Scherbaum W. A. Human monoclonal islet cell antibodies from a patient with insulin-dependent diabetes mellitus reveal glutamate decarboxylase as the target antigen. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8467–8471. doi: 10.1073/pnas.89.18.8467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Riley W. J., Maclaren N. K., Krischer J., Spillar R. P., Silverstein J. H., Schatz D. A., Schwartz S., Malone J., Shah S., Vadheim C. A prospective study of the development of diabetes in relatives of patients with insulin-dependent diabetes. N Engl J Med. 1990 Oct 25;323(17):1167–1172. doi: 10.1056/NEJM199010253231704. [DOI] [PubMed] [Google Scholar]
  24. Scherbaum W. A., Hampl W., Muir P., Glück M., Seissler J., Egle H., Hauner H., Boehm B. O., Heinze E., Banatvala J. E. No association between islet cell antibodies and Coxsackie B, mumps, rubella and cytomegalovirus antibodies in non-diabetic individuals aged 7-19 years. Diabetologia. 1991 Nov;34(11):835–838. doi: 10.1007/BF00408360. [DOI] [PubMed] [Google Scholar]
  25. Seissler J., Hering B., Richter W., Glück M., Yassin N., Bretzel R. G., Boehm B. O., Federlin K., Scherbaum W. A. Antibodies to the M(r) 64,000 (64K) protein in islet cell antibody positive non-diabetic individuals indicate high risk for impaired beta-cell function. Diabetologia. 1992 Jun;35(6):550–554. doi: 10.1007/BF00400483. [DOI] [PubMed] [Google Scholar]
  26. Solimena M., De Camilli P. Autoimmunity to glutamic acid decarboxylase (GAD) in Stiff-Man syndrome and insulin-dependent diabetes mellitus. Trends Neurosci. 1991 Oct;14(10):452–457. doi: 10.1016/0166-2236(91)90044-u. [DOI] [PubMed] [Google Scholar]
  27. Tarn A. C., Thomas J. M., Dean B. M., Ingram D., Schwarz G., Bottazzo G. F., Gale E. A. Predicting insulin-dependent diabetes. Lancet. 1988 Apr 16;1(8590):845–850. doi: 10.1016/s0140-6736(88)91601-7. [DOI] [PubMed] [Google Scholar]
  28. Thivolet C. H., Tappaz M., Durand A., Petersen J., Stefanutti A., Chatelain P., Vialettes B., Scherbaum W., Orgiazzi J. Glutamic acid decarboxylase (GAD) autoantibodies are additional predictive markers of type 1 (insulin-dependent) diabetes mellitus in high risk individuals. Diabetologia. 1992 Jun;35(6):570–576. doi: 10.1007/BF00400486. [DOI] [PubMed] [Google Scholar]
  29. Velloso L. A., Kämpe O., Eizirik D. L., Hallberg A., Andersson A., Karlsson F. A. Human autoantibodies react with glutamic acid decarboxylase antigen in human and rat but not in mouse pancreatic islets. Diabetologia. 1993 Jan;36(1):39–46. doi: 10.1007/BF00399091. [DOI] [PubMed] [Google Scholar]
  30. Wilkin T. J., Schoenfeld S. L., Diaz J. L., Kruse V., Bonifacio E., Palmer J. P. Systematic variation and differences in insulin-autoantibody measurements. Diabetes. 1989 Feb;38(2):172–181. doi: 10.2337/diab.38.2.172. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES