ELISA based on peptide antigens reproducing cross-reactive viral epitopes to detect antibodies in latent autoimmune diabetes in adults vs. type 1 diabetes

MethodsX

Anna Maria Papini¹  Eugenio Ragazzi²  Annunziata Lapolla³  Alessandra Gallo³  Gloria Giovanna Del Vescovo³  Francesca Nuti⁴  Lorenzo Altamore⁴  Feliciana Real-Fernández⁴  Paolo Rovero⁵  Pietro Traldi⁶ 

[1] Corresponding authors.;Department of Pharmaceutical and Pharmacological Sciences, School of Medicine and Surgery, University of Padova, Padova, Italy;Diabetology and Dietetics, Department of Medicine (DIMED), University of Padova, Padova, Italy;Interdepartmental Research Unit of Peptide and Protein Chemistry and Biology, Department of Chemistry “Ugo Schiff”, University of Florence, Sesto Fiorentino, Italy;Interdepartmental Research Unit of Peptide and Protein Chemistry and Biology, Department of Neurosciences, Psychology, Drug Research and Child Health, Section of Pharmaceutical Sciences and Nutraceutics, University of Florence, Sesto Fiorentino, Italy;Istituto di Ricerca Pediatrica, Città della Speranza, Padova, Italy;

关键词: Customized peptide-based solid-phase Enzyme Linked ImmunoSorbent Assay;   

DOI  :  

来源: DOAJ

【 摘 要 】

Diagnosis of Latent Autoimmune Diabetes in Adults (LADA) is based on the adult-age, anti-islet autoantibodies, and temporary insulin-independence. As in Type-1-Diabetes (T1DM), autoimmunity may trigger LADA and enteroviruses-infections can play a role. Anti-human Glutamic-Acid-Decarboxylase (hGAD) autoantibodies are accepted clinical biomarkers, but do not discriminate LADA vs. T1DM. The hypothesis is that protein antigens detecting anti-hGAD antibodies do not expose epitopes specific for different disease forms.We investigated the diagnostic value of autoantibodies in LADA vs. T1DM to peptides of hGAD65/67 isoforms, and Enterovirus-Coxsackie-B4 (CVB4), as antigens sharing the epitope PEVKXK (X: E/T) included in CD8 T-cell CVB4 epitope restricted by diabetes-associated HLA-A2.1. Statistically significant differences of IgM and/or IgG in LADA and T1DM vs. controls were identified. In LADA IgMs to GAD65/67 peptides are diagnostics, IgGs to GAD65/67 peptides correlate with anti-CVB4 peptide antibodies. IgM and/or IgG to all tested peptides can predict LADA, monitoring CVB4 infected patients, improving LADA vs. T1DM stratification. • A customized SP-ELISA based on synthetic peptides Ac-hGAD65(250-273)-NH2 (1), Ac-hGAD67(258-281)-NH2 (2), and Ac-CVB4P2C(28-50)-NH2 (3) is described. • The method was designed to detect specific IgM and/or IgG in LADA, T1DM, vs. controls • Final aim is improvement of LADA vs. T1DM patient stratification.

【 授权许可】

Unknown