High GAD65 autoantibody levels in nondiabetic adults are associated with HLA but not with CTLA-4 or INS VNTR

Abstract. Rolandsson O, Hägg E, Janer M, Rutledge E, Gaur LK, Nilsson M, Hallmans G, Lernmark Å (Umeå University, Umeå, Sweden; Institute for Systems Biology and University of Washington, Seattle, WA, USA). High GAD65 autoantibody levels in nondiabetic adults are associated with HLA but not with CTLA‐4 or INS VNTR. J Intern Med 2003; 253: 447–453. Objectives. To explore the relationship between genetic background and antibody levels in a nondiabetic population. We evaluated if high levels of autoantibodies against the 65 kDa isoform of glutamic acid decarboxylase (GAD65Ab), were associated with high‐risk genes, i.e. HLA, CTLA‐4 and INS VNTR genes. Design and subjects. Seventy‐five (M/F 39/36) subjects exceeding the 95th percentile of GAD65 autoantibody index and 75 age and sex matched subjects below the 95th percentile, randomly selected amongst participants in the Västerbotten Intervention Programme. Methods. The GAD65 Ab were measured in a radioligand‐binding assay. HLA class II typing was performed by an oligoblot hybridization method. CTLA‐4 repeat length was analysed and divided into short forms and long forms. Class I and class III alleles of INS VNTR were detected. Differences in distribution were tested by Pearson chi‐square with Yates correction. Odds ratios (OR) were used to compare groups calculated with Cochran's and Mantel–Haenszel statistics. Results. The DQB1*0201‐DQA1*0501‐DRB1*03 haplotype was increased in subjects with high GAD65Ab levels (P = 0.04). This increase seemed to be explained by a difference in haplotype frequencies amongst men (P = 0.01). Calculating OR showed a significant association between the DQB1*0201‐DQA1*0501‐DRB1*03 haplotype and elevated levels of GAD65Ab in all subjects (OR 2.2, 95% CI 1.02–4.9) as well as in men (OR 4.6, 95% CI 1.3–15.9). There was no association between high levels of GAD65Ab and either INS VNTR or CTLA‐4 polymorphisms. Conclusion. Our study suggests that adult males with the DQB1*0201‐DQA1*0501‐DRB1*03 haplotype tend to develop high GAD65Ab titres. As none of these subjects have developed diabetes these data suggest that HLA may be important in GAD65Ab formation but that additional factors are required for the progression to overt type 1 diabetes.