Summary
The presence of one of the major targets for auto-antibodies in Type 1 (insulin-dependent) diabetes mellitus, the enzyme glutamic acid decarboxylase, was studied in human, rat and mouse pancreatic tissue using immunoprecipitation and immunohistochemical techniques. Immunoprecipitation of glutamic acid decarboxylase was attempted with lysates of [35S]-methionine-labelled rat or mouse pancreatic islets using two different glutamic acid decarboxylase antisera, one mouse monoclonal antibody raised against the 65 kDa isoform of the enzyme, sera from six patients with Type 1 diabetes, one patient with stiff-man syndrome and sera from 19 non-obese diabetic mice. The same sera were used for immunoperoxidase staining of cryosections of human, rat or mouse pancreas. Using patient sera glutamic acid decarboxylase was detected by immunoprecipitations from isolated rat islets but not from islets of five different mouse strains tested, including the non-obese diabetic mouse. When using the non-obese diabetic mouse sera, glutamic acid decarboxylase could not be detected in either rat or mouse tissue. Immunoperoxidase staining demonstrated high levels of glutamic acid decarboxylase in human and rat pancreatic islets but low levels in mouse islets. Direct measurements of enzyme activity showed glutamic acid decarboxylase to be present in mouse islets at a level of about 40% of that in rat islets, and subsequent Western blot analyses indicated that mouse islets express the 67 kDa isoform, whereas in rat islets both the 67 and 65 kDa isoforms are present. The species difference at the level of one of the major islet cell autoantigens in Type 1 diabetes thus indicates that human or rat and mouse islets, respectively, express immunologically distinct forms of glutamic acid decarboxylase and differ in their way of regulating the enzyme production.
Article PDF
Similar content being viewed by others
References
Castano L, Eisenbarth GS (1990) Type I diabetes: a chronic autoimmune disease of human, mouse and rat. Annu Rev Immunol 8: 437–441
Baekkeskoy S, Nielsen JH, Marner B, Bilde T, Ludvigsson J, Lernmark Å (1982) Autoantibodies in newly diagnosed diabetic children immunoprecipitate human pancreatic islet cell proteins. Nature (Lond) 298: 167–169
Baekkeskov S, Aanstoot HJ, Christgau S et al. (1990) Identification of the 64K autoantigen in insulin-dependent diabetes as the GABA-synthesizing enzyme glutamic acid decarboxylase. Nature (Lond). 347: 151–156
Lernmark Å, Freedman ZR, Hoffman C et al. (1978) Islet cell-surface antibodies in juvenile diabetes mellitus. N Engl J Med 299: 375–380
Thomas NM, Ginsberg-Fellner F, McEvoy RC (1990) Strong association between diabetes and displacement of mouse anti-rat insulinoma cell monoclonal antibody by human serum in vitro. Diabetes 39: 1203–1211
Kolb H (1987) Mouse models of insulin dependent diabetes: low-dose streptozotocin induced-diabetes and nonobese diabetic (NOD) mice. Diabetes Metab Rev 3: 751–778
Atkinson MA, McLaren NK (1988) Autoantibodies in nonobese diabetic mice immunoprecipitate 64,000 Mr islet antigen. Diabetes 37: 845–850
Kämpe O, Andersson A, Björk E, Hallberg A, Karlsson FA (1989) High-glucose stimulation of 64,000 Mr islet cell autoantigen expression. Diabetes 38: 1326–1328
Kämpe O, Hallberg A, Björk E, Andersson A, Karlsson FA (1990) Human diabetic sera react with rat but not mouse islet cells. Immunology of Diabetes, 10th International Workshop, Jerusalem (Abstract)
Oertel WH, Schmechel DE, Mugnaini E, Tappaz ML, Kopin IJ (1981) Immunocytochemical localization of glutamate decarboxylase in rat cerebellum with a new antiserum. Neuroscience 6: 2715–2735
Erlander MG, Tillakaratne NJK, Feldblum S, Patel N, Tobin AJ (1991) Two genes encode distinct glutamate decarboxylase with different responses to pyridoxal phosphate. Neuron 7: 91–100
Howell SL, Taylor KW (1968) Potassium ions and the secretion of insulin by islets of Langerhans incubated in vitro. Biochem J 108: 17–24
Strandell E, Eizirik DL, Sandler S (1990) Reversal of β5-cell suppression in vitro pancreatic islets isolated from non obese diabetic mice during the phase preceding insulin-dependent diabetes mellitus. J Clin Invest 85: 1944–1950
Andersson A (1978) Isolated mouse pancreatic islets in culture: effects of serum and different culture media on the insulin production of the islets. Diabetologia 14: 397–404
Blobel G, Dobberstein B (1975) Transfer of proteins across membranes IL Reconstitution of functional rough microsomes from heterologous components. J Cell Biol 67: 852–862
Molinoff PB, Kravitz EA (1968) The metabolism of gamma-aminobutyric acid (GABA) in the lobster nervous system — glutamic decarboxylase. J Neurochem 15:391–409
Christie MR, Brown TJ, Cassidy D (1992) 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 35: 380–384
Winqvist O, Karlsson FA, Kämpe O (1992) 21-hydroxylase, a major autoantigen in idiopathic Addisons's disease. Lancet 339: 1559–1562
Kaufman DL, Houser CR, Tobin AJ (1991) Two forms of the gamma-amino butyric acid synthetic enzyme glutamate decarboxylase have distinct intraneuronal distributions and cofactor interactions. J Neurochem 56:720–723
Chang YC, Gottlieb DI (1988) Characterization of the proteins purified with monoclonal antibodies to glutamic acid decarboxylase. J Neurosci 8:2123–2130
Andersson A, Korsgren O, Naeser P (1990) DNA replication in transplanted and endogenous pancreatic islets of obese-hyperglycemic mice at different stages of the syndrome. Metabolism 38:974–978
Gilon P, Tappaz M, Remade C (1991) Localization of GAD-like immunoreactivity in pancreas and stomach of the rat and mouse. Histochemistry 96:355–365
Okada Y, Taniguchi H, Baba S (1982) High concentration of GABA in the pancreatic islets with special emphasis on β-cells. Excerpta Medica, Amsterdam, pp 379–386
Karlsen AE, Hagopian WA, Grubin CE et al. (1991) Cloning and primary structure of a human islet isoform of glutamic acid decarboxylase from chromosome 10. Proc Natl Acad Sci USA. 88: 8337–8341
Björk E, Kämpe O, Karlsson FA et al. Glucose regulation of the autoantigen GAD 65 in human pancreatic islets. J Clin Endocrinol Metabol (in press)
Gerling I, Chatterjee NK (1990) Autoantigen (64,000) expression in Coxsackie virus B4-induced experimental diabetes. Curr Top Microbiol Immunol 156:55–62
Gerling I, Nejman C, Chatterjee NK (1988) Effect of Coxsakie virus B4 infection in mice on expression of 64,000 Mr autoantigen and glucose sensitivity of islets before development of hyperglycemia. Diabetes 37:1419–1425
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Velloso, L.A., Kämpe, O., Eizirik, D.L. et al. Human autoantibodies react with glutamic acid decarboxylase antigen in human and rat but not in mouse pancreatic islets. Diabetologia 36, 39–46 (1993). https://doi.org/10.1007/BF00399091
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00399091