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Horm Metab Res 2011; 43(7): 483-488
DOI: 10.1055/s-0031-1275705
Original Basic

© Georg Thieme Verlag KG Stuttgart · New York

Recognition of Ii-Key/MHC Class II Epitope Hybrids Derived from Proinsulin and GAD Peptides by T Cells in Type 1 Diabetes

M. Vadacca1 , M. G. Valorani2 , 3 , E. von Hofe4 , N. L. Kallinteris4 , R. Buzzetti5 , P. Pozzilli1 , 2
  • 1Department of Endocrinology and Diabetes, University Campus Bio-Medico, Rome, Italy
  • 2Bart's & the London School of Medicine & Dentistry, Centre of Diabetes, Institute of Cell and Molecular Science, Queen Mary University of London, UK
  • 3Immunology Research Department, Livio Patrizi Foundation, Research Group BIOS, Rome, Italy
  • 4Antigen Express Inc., Worcester, MA, USA
  • 5Department of Clinical Science, University Sapienza, Rome, Italy
Further Information

Publication History

received 10.09.2010

accepted 23.03.2011

Publication Date:
21 April 2011 (online)

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Abstract

In order to determine whether the Ii-Key technology can enhance the presentation of specific epitopes associated with type 1 diabetes, we have designed and synthesized a series of Ii-Key/proinsulin and GAD epitope hybrid peptides. Peptides of proinsulin and GAD shown to be recognized by CD4+ T cells of type 1 diabetes patients have been selected from the literature and modified with Ii-Key. A total of 23 Caucasian type 1 diabetes subjects and 17 normal subjects as controls were included in the study. Reactive T cells were identified using an IFN-γ ELISPOT assay. We selected 5 proinsulin and 5 GAD epitopes. Regarding the activity of the proinsulin Ii-Key hybrids, 3 out of 15 patients (20%) demonstrated a positive response to one or more Ii-Key hybrid peptides compared to no responders in the control subjects. Two out of 8 patients demonstrated a positive response to one or more Ii-Key/GAD65 hybrids. Proinsulin Ii-Key hybrids and peptides were recognized only by DR3/DR4 0302+ve diabetic patients. Control subjects showed no detectable response to stimulation with Ii-Key hybrids or peptides, neither for proinsulin nor GAD65. We have now shown that the use of Ii-Key-modified MHC class II epitopes, derived from proteins associated with insulin-secreting cells, can detect the presence of specifically activated CD4+ T helper cells with greater sensitivity than unmodified epitopes in the standard ELISPOT assay. The use of these technologies may be of use in identifying patients at the earliest stages of type 1 diabetes.

Key words

Ii-Key hybrid - type 1 diabetes - MHC class II

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Correspondence

Dr. M VadaccaMD, PhD 

University Campus Bio-Medico

Via Alvaro del Portillo 21

00128 Rome

Italy

Phone: +39/062/254 11

Fax: +39/062/254 14 56

Email: m.vadacca@unicampus.it

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