Radioimmunoassays for glutamic acid decarboxylase (GAD65) and GAD65 autoantibodies using 35S or 3H recombinant human ligands

Falorni, Alberto; Örtqvist, Eva; Persson, Bengt; Lernmark, Åke

doi:10.1016/0022-1759(95)00139-2

Cited by 184 publications

(151 citation statements)

References 26 publications

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“…The serum samples of 2157 subjects were analysed for GAD65Ab in a radioligand-binding assay described in detail elsewhere [10]. In short, human recombinant GAD65 was produced by in vitro transcription/translation and labelled with 35S-methionine [12].…”

Section: Methodsmentioning

confidence: 99%

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Glutamate decarboxylase (GAD65) and tyrosine phosphatase-like protein (IA-2) autoantibodies index in a regional population is related to glucose intolerance and body mass index

et al. 1999

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The clinical diagnosis of diabetes mellitus is based on specific criteria of blood glucose concentrations [1]. After diagnosis of the disease, clinical criteria are used to classify diabetes into Type I (insulin-dependent) or Type II (non-insulin-dependent) diabetes mellitus. The classification of diabetes is mostly unambiguous in people younger than 20 years of age and in older patients who predominately have Type II diabetes. It is widely recognised, however, that Type I diabetes can develop at any age which complicates classification in the older patient. Type I diabetes in children and young adults is strongly associated with the presence of autoantibodies against the Mr 65 kD isoform of glutamate decarboxylase (GAD65) [2] and IA-2 (ICA512), a tyrosine phosphatase-like transmembrane protein [3,4]. The diagnostic sensitivity of GAD65Ab has been reported to be about 70±80 % and the diagnostic specificity 99 % in such patients [5]. Furthermore, in newly diagnosed Type Diabetologia (1999) Results. We found 23 of 2157 (1.1 %) and 17 of 2152 (0.8 %) subjects exceeded the 99th centile of GAD65 autoantibody index and IA-2 autoantibody index, respectively. In 18 subjects with diabetic oral glucose tolerance test, GAD65 autoantibody concentrations were higher than in those with normal oral glucose tolerance test (p = 0.02). Subjects with IGT (n = 416) and diabetes (n = 18), i. e. abnormal OGTT (n = 434), had a higher IA-2Ab index compared with those with normal OGTT (p = 0.008). A stepwise multiple logistic regression test showed that the odds ratios for subjects in the highest BMI group to exceed the 95th or 99th GAD65 autoantibody centile were 3.6 (CI 1.4±8.9) and 17.6 (CI 2.6±121.6), respectively. Conclusion/interpretation. GAD65 and IA-2 autoantibodies, are associated with impaired or diabetic glucose tolerance in an adult regional population. This observation together with the association between GAD65 autoantibody concentrations and body mass index indicate a possible relation between islet autoimmunity and beta-cell function abnormalities with obesity and insulin resistance. [Diabetologia (1999)

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Section: Methodsmentioning

confidence: 99%

“…We have taken advantage of the standardised, highly sensitive and specific test for GAD65Ab and IA-2Ab using in vitro transcribed and translated human recombinant GAD65 and IA-2 [10,11] and a population-based investigation of 2157 subjects being either 30, 40, 50 or 60 years of age.…”

Section: : 555±559]mentioning

confidence: 99%

Glutamate decarboxylase (GAD65) and tyrosine phosphatase-like protein (IA-2) autoantibodies index in a regional population is related to glucose intolerance and body mass index

et al. 1999

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“…DKA was defined as described previously by us (3). GAD65 and insulinomaassociated protein-2 autoantibodies were measured in sera as reported previously (29,30,31), using normal ranges established for each ethnic group (3). Patients were classified as ␤ϩ or ␤Ϫ by the presence or absence of ␤-cell functional reserve, as measured within 2 weeks after resolution of initial presentation with DKA and after the 12-month follow-up.…”

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confidence: 99%

HLA Class II Alleles Specify Phenotypes of Ketosis-Prone Diabetes

et al. 2008

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OBJECTIVE -Ketosis-prone diabetes (KPD) comprises four subgroups based on the presence or absence of ␤-cell autoantibodies (Aϩ or AϪ) and ␤-cell functional reserve (␤ϩ or ␤Ϫ). Genetic factors could contribute to their distinctive phenotypes. Our aim was to specify the role of HLA class II alleles associated with susceptibility or resistance to autoimmune type 1 diabetes in determining KPD phenotypes.RESEARCH DESIGN AND METHODS -A total of 185 adults presenting with diabetic ketoacidosis were followed longitudinally for a mean of 5.5 years, with measurements of autoantibodies, ␤-cell functional reserve, insulin sensitivity, and insulin requirement. Frequencies of susceptibility and resistance alleles at HLA DQA1, DQB1, and DRB1 loci were correlated with clinical and phenotypic features of KPD subgroups and compared with those of ethnic-specific population control subjects.RESULTS -Susceptibility alleles were more frequent (P Ͻ 0.0001) in the two Aϩ than the two AϪ KPD subgroups; in the latter, the frequency was no greater than in population control subjects (except for DQB1*0302). Susceptibility alleles differentiated the two clinically similar ␤Ϫ subgroups (more frequent in Aϩ␤Ϫ than AϪ␤Ϫ KPD; P Ͻ 0.01). Resistance alleles were more frequent in the two ␤ϩ than the two ␤Ϫ KPD subgroups (P Ͻ 0.01). The frequencies of certain susceptibility (e.g., DQB1*02) and resistance (DQB1*0602) alleles were higher in African-American AϪ␤ϩ KPD patients than in African-American control subjects. DQB1*0302 was more frequent in all KPD subgroups compared with control subjects.CONCLUSIONS -HLA class II alleles associated with susceptibility or resistance to autoimmune type 1 diabetes help specify the four subgroups of KPD. Inheritance of these alleles may influence long-term ␤-cell functional reserve.

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“…Preparation of IgG subclass-specific Sepharose for the use in the biotin/SPBA -immobilized Preparation of recombinant 35 S-methionine-labelled GAD65 and incubation with human plasma have been described in detail elsewhere [10,11]. The recombinant GAD65 was diluted in wash buffer [10 000 counts per minute (cpm)/ well] and incubated with plasma overnight at +4°C during agitation.…”

Section: Subjectsmentioning

confidence: 99%

Determination of glutamic acid decarboxylase antibodies (GADA) IgG subclasses − comparison of three immunoprecipitation assays (IPAs)

Hillman

Törn

Landin‐Olsson

2007

Clinical and Experimental Immunology

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Radioimmunoassays for glutamic acid decarboxylase (GAD65) and GAD65 autoantibodies using 35S or 3H recombinant human ligands

Cited by 184 publications

References 26 publications

Glutamate decarboxylase (GAD65) and tyrosine phosphatase-like protein (IA-2) autoantibodies index in a regional population is related to glucose intolerance and body mass index

Glutamate decarboxylase (GAD65) and tyrosine phosphatase-like protein (IA-2) autoantibodies index in a regional population is related to glucose intolerance and body mass index

HLA Class II Alleles Specify Phenotypes of Ketosis-Prone Diabetes

Determination of glutamic acid decarboxylase antibodies (GADA) IgG subclasses − comparison of three immunoprecipitation assays (IPAs)

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