title: Islet amyloid polypeptide is a target antigen for diabetogenic CD4+ T cells.
abstract: OBJECTIVE
  To investigate autoantigens in β-cells, we have used a panel of pathogenic T-cell clones that were derived from the NOD mouse. Our particular focus in this study was on the identification of the target antigen for the highly diabetogenic T-cell clone BDC-5.2.9.
  
  
  METHODS
  To purify β-cell antigens, we applied sequential size exclusion chromatography and reverse-phase high-performance liquid chromatography to membrane preparations of β-cell tumors. The presence of antigen was monitored by measuring the interferon-γ production of BDC-5.2.9 in response to chromatographic fractions in the presence of NOD antigen-presenting cells. Peak antigenic fractions were analyzed by ion-trap mass spectrometry, and candidate proteins were further investigated through peptide analysis and, where possible, testing of islet tissue from gene knockout mice.
  
  
  RESULTS
  Mass-spectrometric analysis revealed the presence of islet amyloid polypeptide (IAPP) in antigen-containing fractions. Confirmation of IAPP as the antigen target was demonstrated by the inability of islets from IAPP-deficient mice to stimulate BDC-5.2.9 in vitro and in vivo and by the existence of an IAPP-derived peptide that strongly stimulates BCD-5.2.9.
  
  
  CONCLUSIONS
  IAPP is the target antigen for the diabetogenic CD4 T-cell clone BDC-5.2.9.
authors: Integrated Department of Immunology, University of Colorado Denver School of Medicine and National Jewish Health, Denver, Colorado, USA.
authors: Thomas
authors: T
authors: Delong
authors: Rocky L
authors: RL
authors: Baker
authors: Nichole
authors: N
authors: Reisdorph
authors: Richard
authors: R
authors: Reisdorph
authors: Roger L
authors: RL
authors: Powell
authors: Michael
authors: M
authors: Armstrong
authors: Gene
authors: G
authors: Barbour
authors: Brenda
authors: B
authors: Bradley
authors: Kathryn
authors: K
authors: Haskins
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D001324:Autoantigens / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D058228:Islet Amyloid Polypeptide / Q000276:immunology*; D007515:Islets of Langerhans / Q000276:immunology*; D013058:Mass Spectrometry; D051379:Mice; D016688:Mice, Inbred NOD
chemical_list: D001324:Autoantigens; D058228:Islet Amyloid Polypeptide