Single cell assessment of allergen-specific T cell responses with MHC class II peptide tetramers: Methodological aspects

Erik Wambre, Laurence Van Overtvelt, Bernard Maillère, Robert Humphreys, Eric Von Hofe, Lydia Ferhat, Didier Ebo, Philippe Moingeon

Research output: Contribution to journalArticlepeer-review

31 Scopus citations


Background: We report herein critical methodological principles for assessing, at a single cell level, allergen-specific T cell responses using MHC class II peptide tetramers. Methods: We developed MHC class II peptide tetramers to monitor T cell responses against the immunodominant Bet v 1 141-155 peptide in individuals with either an HLA-DRB1*0101, DRB1*0401 or DRB1*1501 background. In vitro stimulation was performed with serially truncated versions of the Bet v 1141-155 epitope chemically conjugated to the Ii-Key peptide. Results: Identification of Bet v 1141-155 as a high-affinity epitope for multiple HLA-DRB1 allotypes led to the development of corresponding tetramers detecting Bet v 1141-155-specific T cells with a high specificity and sensitivity. Stimulation with Bet v 1141-155 Ii-Key conjugate peptides is the most efficient procedure to expand Bet v 1141-155-specific CD4+ T cells, allowing to detect such cells in both allergic and healthy individuals. MHC class II Bet v 1141-155 tetramer-positive T cells produce IFN-γ and IL-10 in healthy individuals, and IL-5 in allergic patients. Frequencies of Bet v 1-specific CD4+ T cells circulating in the blood of allergic or nonallergic individuals range from approximately 10-5 to 10 -3 CD4+ T cells, outside or within the pollen season, respectively. Conclusions: MHC class II peptide tetramers are valuable tools to assess allergen-specific T cell responses, both qualitatively and quantitatively. Selection of a high-affinity T cell epitope, as well as optimization of in vitro stimulation conditions to expand rare T cell progenitors are critical success factors in those analyses.

Original languageEnglish
Pages (from-to)99-112
Number of pages14
JournalInternational Archives of Allergy and Immunology
Issue number2
StatePublished - May 2008
Externally publishedYes


  • Bet v 1
  • MHC class II tetramers
  • T cell tolerance


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