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 ((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)),((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)),((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)):

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 ((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))