title: Crohn's disease-specific anti-CUZD1 pancreatic antibodies are absent in ruminants with paratuberculosis.
abstract: BACKGROUND
  Pancreatic autoantibodies (PABs) specifically recognizing GP2 and/or CUZD1 are present in more than 35% of patients with Crohn's disease (CrD). We have recently provided evidence of the presence of GP2-specific PABs in ruminants with paratuberculosis (ptb), a Mycobacterium avium paratuberculosis (MAP)-induced disease resembling CrD.
  
  
  OBJECTIVE
  To assess whether anti-CUZD1 antibodies are also present in ruminants with ptb.
  
  
  METHODS
  A total of 110 samples (73 cattle/37 sheep) were studied including 40 with ptb (24 cattle/16 sheep; 20 anti-GP2 antibody pos) and 70 without ptb (49 cattle/21 sheep; 10 anti-GP2 antibody pos). The samples were pre-characterized for anti-MAP and anti-GP2 antibodies by ELISA. Evidence of MAP was confirmed by PCR. Anti-CUZD1 antibody testing was performed by indirect immunofluorescence (IIF) based on transfected HEK293 cells expressing CUZD1. Anti-sheep or anti-cattle specific antisera were used as revealing antibodies.
  
  
  RESULTS
  None of the ruminant sera had anti-CUZD1 antibodies by IIF testing at dilutions varying from 1/10 to 1/160. Methodological flaws were prevented by a series of tests. Control sera from anti-CUZD1 positive CrD samples have shown anti-CUZD1 antibody reactivity at various concentrations. Antibody reactivity to GP2-expressing HEK293 cells has confirmed the reactivity to GP2 in ruminant sera found positive for anti-GP2 antibodies by ELISA.
  
  
  CONCLUSIONS
  The present study has found no evidence of anti-CUZD1 PABs in MAP-induced ptb. Our findings indicate that the induction of CUZD1-specific PABs is unrelated to MAP infection and that the mechanisms responsible for the loss of tolerance to GP2 and CUZD1 are probably quite distinct.
authors.affiliation: Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece; Department of Rheumatology, Faculty of Medicine, School of Health Sciences, University of Thessaly, Biopolis, 45000 Larissa, Greece. Electronic address: liaskosch@med.uth.gr.
authors.forename: Christos
authors.initials: C
authors.lastname: Liaskos
authors.affiliation: Department of Animal Production, Technological Educational Institute of Thessaly, Larissa 41110, Greece. Electronic address: spyrou@teilar.gr.
authors.forename: Vassiliki
authors.initials: V
authors.lastname: Spyrou
authors.affiliation: Department of Medicine, Faculty of Veterinary Medicine, University of Thessaly, Karditsa 43100, Greece. Electronic address: lathan@vet.uth.gr.
authors.forename: Labrini V
authors.initials: LV
authors.lastname: Athanasiou
authors.affiliation: Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece. Electronic address: kleliaorf_bet@yahoo.gr.
authors.forename: Timoklia
authors.initials: T
authors.lastname: Orfanidou
authors.affiliation: Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece. Electronic address: mavropoulos_thanos@hotmail.com.
authors.forename: Athanasios
authors.initials: A
authors.lastname: Mavropoulos
authors.affiliation: Division of Transplantation Immunology and Mucosal Biology, King's College London School of Medicine at King's College Hospital, London SE5 9RJ, UK. Electronic address: eirigopoulou@med.uth.gr.
authors.forename: Eirini I
authors.initials: EI
authors.lastname: Rigopoulou
authors.affiliation: Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, University of Thessaly, Karditsa 43100, Greece. Electronic address: gsamir@vet.uth.gr.
authors.forename: Georgios S
authors.initials: GS
authors.lastname: Amiridis
authors.affiliation: The Zabludowicz Center for Autoimmune Diseases, Sheba Medical Center, Tel-Hashomer, Israel. Electronic address: shoenfel@post.tau.ac.il.
authors.forename: Yehuda
authors.initials: Y
authors.lastname: Shoenfeld
authors.affiliation: Department of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Thessaly, 43100 Karditsa, Greece. Electronic address: billinis@vet.uth.gr.
authors.forename: Charalambos
authors.initials: C
authors.lastname: Billinis
authors.affiliation: Cellular Immunotherapy and Molecular Immunodiagnostics, Biomedical Section, Institute for Research and Technology Thessaly (IReTeTh), ITE, Larissa 41222, Greece; Department of Rheumatology, Faculty of Medicine, School of Health Sciences, University of Thessaly, Biopolis, 45000 Larissa, Greece; Division of Transplantation Immunology and Mucosal Biology, King's College London School of Medicine at King's College Hospital, London SE5 9RJ, UK. Electronic address: bogdanos@med.uth.gr.
authors.forename: Dimitrios P
authors.initials: DP
authors.lastname: Bogdanos
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000097:blood*; D002417:Cattle; D003424:Crohn Disease / Q000276:immunology*; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology*; D010179:Pancreas / Q000276:immunology*; D010283:Paratuberculosis / Q000097:blood*; D012418:Ruminants; D012756:Sheep
chemical_list: D001323:Autoantibodies; C488236:CUZD1 protein, human; D008565:Membrane Proteins