Human in vitro assays

Mechanisms of allergy and tolerance in humans are investigated at the T cell and IgE level. Long-term and short-term T cell culture-systems have been set-up to measure allergen-specific T cell responses towards cow’s milk, peanut and other allergens. Furthermore, B cell responses at the level of immunoglobulin subtypes (IgE, IgG1, IgG4) are studied.

T and B cell responses have been analyzed in various groups of subjects 1,2,3:

Allergic individuals: have specific IgE towards the allergen of interest and clinical symptoms

  • Sensitized non-allergic individuals: have specific IgE towards the allergen of interest but no clinical symptoms
  • Atopic individuals: IgE towards other allergens, but not to the allergen of interest
  • Non-atopic individuals: no family history of atopy, no elevated IgE

In vitro assays for functional IgE reactivity are available:

  • RBL assay: rat mast cell line transfected with the human IgE-receptor
  • Basophil degranulation assay: human basophil degranulation measured by histamine release or CD63 expression 4

  1. Tiemessen MM, Van Ieperen-Van Dijk AG, Bruijnzeel-Koomen CA, Garssen J, Knol EF, Van Hoffen E. Cow’s milk-specific T-cell reactivity of children with and without persistent cow’s milk allergy: key role for IL-10. J Allergy Clin Immunol 2004;113(5):932-9 []
  2. Ruiter B, TrĂ©goat V, M’rabet L, Garssen J, Bruijnzeel-Koomen CA, Knol EF, Hoffen E. Characterization of T cell epitopes in alphas1-casein in cow’s milk allergic, atopic and non-atopic children. Clin Exp Allergy 2006;36(3):303-10 []
  3. Ruiter B, Knol EF, van Neerven RJ, Garssen J, Bruijnzeel-Koomen CA, Knulst AC, van Hoffen E. Maintenance of tolerance to cow’s milk in atopic individuals is characterized by high levels of specific immunoglobulin G4. Clin Exp Allergy 2007;37(7):1103-10 []
  4. Koppelman SJ, Wensing M, Ertmann M, Knulst AC, Knol EF. Relevance of Ara h1, Ara h2 and Ara h3 in peanut-allergic patients, as determined by immunoglobulin E Western blotting, basophil-histamine release and intracutaneous testing: Ara h2 is the most important peanut allergen. Clin Exp Allergy 2004;34(4):583-90 []