Simultaneous detection of circulating autoreactive CD8+ T-cells specific for different islet cell-associated epitopes using combinatorial MHC multimers

Jurjen H. Velthuis, Wendy W. Unger, Joana R. F. Abreu, Gaby Duinkerken, Kees Franken, Mark Peakman, Arnold H. Bakker, Sine Reker Hadrup, Bart Keymeulen, Jan Wouter Drijfhout, Ton N. Schumacher, Bart O. Roep

Research output: Contribution to journalJournal articleResearchpeer-review


OBJECTIVE - Type 1 diabetes results from selective T-cell-mediated destruction of the insulin-producing β-cells in the pancreas. In this process, islet epitope-specific CD8+ T-cells play a pivotal role. Thus, monitoring of multiple islet-specific CD8+ T-cells may prove to be valuable for measuring disease activity, progression, and intervention. Yet, conventional detection techniques (ELISPOT and HLA tetramers) require many cells and are relatively insensitive. RESEARCH DESIGN AND METHODS - Here, we used a combinatorial quantum dot major histocompatibility complex multimer technique to simultaneously monitor the presence of HLA-A2 restricted insulin B10-18, prepro-insulin (PPI)15-24, islet antigen (IA)-2797-805, GAD65114-123, islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) 265-273, and pre-pro islet amyloid polypeptide (ppIAPP) 5-13-specific CD8+ T-cells in recent-onset diabetic patients, their siblings, healthy control subjects, and islet cell transplantation recipients. RESULTS - Using this kit, islet autoreactive CD8+ T-cells recognizing insulin B10-18, IA-2 797-805, and IGRP265-273 were shown to be frequently detectable in recent-onset diabetic patients but rarely in healthy control subjects; PPI15-24 proved to be the most sensitive epitope. Applying the "Diab-Q-kit" to samples of islet cell transplantation recipients allowed detection of changes of autoreactive T-cell frequencies against multiple islet cell-derived epitopes that were associated with disease activity and correlated with clinical outcome. CONCLUSIONS - A kit was developed that allows simultaneous detection of CD8+ T-cells reactive to multiple HLA-A2-restricted β-cell epitopes requiring limited amounts of blood, without a need for in vitro culture, that is applicable on stored blood samples. © 2010 by the American Diabetes Association.
Original languageEnglish
Issue number7
Pages (from-to)1721-1730
Number of pages10
Publication statusPublished - 2010
Externally publishedYes


  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • CD8 antigen
  • epitope
  • glutamate decarboxylase 65[114-123]
  • HLA A2 antigen
  • insulin B[10-18]
  • islet antigen 2[797-805]
  • islet specific glucose 6 phosphate catalytic subunit related protein[5-13]
  • major histocompatibility antigen
  • prepro islet amyloid polypeptide[5-13]
  • preproinsulin[15-24]
  • quantum dot
  • unclassified drug
  • adolescent
  • article
  • blood sampling
  • CD8+ T lymphocyte
  • child
  • controlled study
  • diabetic patient
  • disease activity
  • disease course
  • enzyme linked immunospot assay
  • human
  • human cell
  • insulin dependent diabetes mellitus
  • major clinical study
  • major histocompatibility complex
  • pancreas islet cell
  • pancreas islet transplantation
  • priority journal
  • Adolescent
  • CD8-Positive T-Lymphocytes
  • Child
  • Child, Preschool
  • Diabetes Mellitus, Type 1
  • Epitopes
  • Female
  • Flow Cytometry
  • HLA-A Antigens
  • Humans
  • Infant
  • Insulin-Secreting Cells
  • Islets of Langerhans
  • Islets of Langerhans Transplantation
  • Major Histocompatibility Complex
  • Male
  • Statistics, Nonparametric


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