title: T-cell mediated autoimmunity to the insulinoma-associated protein 2 islet tyrosine phosphatase in type 1 diabetes mellitus.
abstract: The target molecules of the T-cell response in type 1 diabetes, despite their pathogenic importance, remain largely uncharacterized, especially in humans. Interestingly, molecules such as insulin and glutamic acid decarboxylase (GAD) have been shown to be a target not only of autoantibodies, but also of autoreactive T-lymphocytes both in man and in the non-obese diabetic (NOD) mouse. In the present study we aimed to determine the existence of a specific T-cell response towards the insulinoma-associated protein 2 (IA-2) islet tyrosine phosphatase, a recently identified autoantigen which is the target of autoantibodies strongly associated with diabetes development. Human recombinant IA-2 produced in Escherichia coli, was tested for its reactivity with peripheral blood lymphocytes obtained from 16 newly diagnosed type 1 diabetic patients and from 25 normal controls, 15 of whom were HLA-DR-matched. A T-cell proliferation assay was performed in triplicate employing freshly isolated cells in the absence or in the presence of the antigen to be tested (at two different concentrations: 2 microg/ml and 10 microg/ml). A specific T-cell proliferation (defined as a stimulation index (S.I.) >/=3) was observed against IA-2 used at a concentration of 10 microg/ml (but not of 2 microg/ml) in 8/16 diabetic patients, in 1/15 HLA-DR-matched control subjects (P<0.01 by Fisher exact test) and in 0/10 of the remaining normal individuals. A statistically significant difference (P<0.003 by Mann-Whitney U test) was also observed in S.I. values between patients (3.1+/-1.4) and HLA-DR-matched controls (1.7+/-0.54) employing IA-2 at a concentration of 10 microg/ml. However, when IA-2 was used at a concentration of 2 microg/ml, the difference in S. I. between patients (1.65+/-0.8) and controls (1.0+/-0.3) did not reach statistical significance. In conclusion, these data show the presence of a specific, dose-dependent T-lymphocyte response against the IA-2 islet tyrosine phosphatase at the onset of type 1 diabetes. Consequently, this molecule appears to be a target not only at the B-lymphocyte but also at the T-lymphocyte level, reinforcing the potential pathogenic role of this autoantigen in the islet destructive process.
authors.affiliation: Department of Endocrinology, University 'La Sapienza', Rome, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Dotta
authors.forename: S
authors.initials: S
authors.lastname: Dionisi
authors.forename: V
authors.initials: V
authors.lastname: Viglietta
authors.forename: C
authors.initials: C
authors.lastname: Tiberti
authors.forename: M C
authors.initials: MC
authors.lastname: Matteoli
authors.forename: M
authors.initials: M
authors.lastname: Cervoni
authors.forename: C
authors.initials: C
authors.lastname: Bizzarri
authors.forename: G
authors.initials: G
authors.lastname: Marietti
authors.forename: M
authors.initials: M
authors.lastname: Testi
authors.forename: G
authors.initials: G
authors.lastname: Multari
authors.forename: L
authors.initials: L
authors.lastname: Lucentini
authors.forename: U
authors.initials: U
authors.lastname: Di Mario
mesh_terms: D000293:Adolescent; D000328:Adult; D001324:Autoantigens / Q000276:immunology*; D015551:Autoimmunity / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D004587:Electrophoresis, Agar Gel; D005260:Female; D005968:Glutamate Decarboxylase / Q000276:immunology; D006684:HLA-DR Antigens / Q000032:analysis; D006650:Histocompatibility Testing; D006801:Humans; D007328:Insulin / Q000276:immunology; D008297:Male; D008565:Membrane Proteins / Q000276:immunology*; D016133:Polymerase Chain Reaction; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases / Q000276:immunology*; D011863:Radioimmunoassay; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D011994:Recombinant Proteins; D012588:Scintillation Counting; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D001324:Autoantigens; D006684:HLA-DR Antigens; D007328:Insulin; D008565:Membrane Proteins; D011994:Recombinant Proteins; C516862:PTPRN protein, human; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D017027:Protein Tyrosine Phosphatases; C516863:Ptprn protein, mouse; D054635:Receptor-Like Protein Tyrosine Phosphatases, Class 8; D005968:Glutamate Decarboxylase