Ugotavljanje bolnikove kompletne specifične IgE alergogene aktivnosti z uporabo kombinacije rekombinantnih alergenov in multiparameterske analize aktivacije bazofilcev (Slovene)

Evaluation of the allergenic activity of allergic patients IgE using recombinant allergens and multiplex basophil activation testing   More than 25% of the population suffers from IgE-associated allergy. Currently, the in vitro diagnosis of allergic diseases is based on measurement of IgE reactivity to allergens, which basically only provides information about the presence and/or concentration of specific IgE in patient’s sera. Unfortunately, this standard approach does not give any information relating to biologic relevance i.e. allergenic activity of the IgE antibodies measured in sera and thus the clinical relevance of allergen-specific IgE is very limited. Importantly, a significant number of individuals with allergen specific IgE do not develop symptoms. Furthermore, allergic diseases range from mild to systemic and severe allergic reactions. Therefore, establishment of the clinical relevance of patient’s IgE is a very important issue. Recently two distinct approaches for the in vitro evaluation of the clinical relevance of IgE antibodies were developed: one is the use of recombinant allergens and the second one is the use of the basophil activation test (BAT). In this collaborative project, a group with profound expertise in producing defined recombinant allergens (Austrian partner from the Medical University Vienna: Prof. Rudolf Valenta) will work together with Dr. Peter Korosec from the University Clinic in Golnik, Slovenia, a world renowned expert in basophil activation testing. The allergic disease model will be house dust mite (HDM) which is one of the most important indoor allergen source. The combined expertise of the two groups will allow to incorporate recombinant allergens in BAT (basophil activation testing) testing and to develop a novel platform of recombinant BAT in three steps. First, complete basophil CD63 log dose response curves for an extensive panel of recombinant HDM allergens (rDer p 2, 4, 5, 7, 10, 11, 14, 15, 18, 21 and 23) and nDer p 1 will be established. In a second step HDM recombinant allergens will be bound to different fluorophores which could be used in flow cytometry and thus in BAT testing. The third step will be the most advanced as it will incorporate the second step using different recombinant allergens bound to fluorophores in combination with the multiplex BAT allergenic activity measurement. The proposed multiplex BAT assay should overcome disadvantages of current in vitro IgE measurement approaches, as will not show only to which components patients are IgE sensitized, but also what is the allergenic activity of the individual allergen-specific IgE antibodies. The proposed multiplex methodology should make basophil testing applicable for daily diagnostic allergy testing.

The in vitro diagnosis of allergic diseases is currently based on measurement of IgE reactivity to allergens, which basically provides information about the presence and/or concentration of specific IgE in patient’s sera. Unfortunately, this standard approach does give only limited information relating to biologic relevance i.e. allergenic activity of the IgE antibodies. Furthermore, allergic diseases range from mild to systemic and severe allergic reactions. Therefore, establishment of the allegenic activity of patient’s IgE is a very important issue. Recently two approaches for the in vitro evaluation of the clinical relevance of IgE antibodies were developed: one is the use of recombinant allergens and the second one is the use of the basophil activation test (BAT). In this project we combine the methodology and technology of recombinant allergens and basophil activation test with the aim to develop novel diagnostic and research platforms. We established and characterized IgE allergenic activity on different clinical and experimental models of allergic diseases and further explored differences in effector cell function between basophils and mast cell during systemic allergic reaction - anaphylaxis. The major part of this project was already published, including two papers in Journal of Allergy and Clinical Immunology (JACI) with impact factor 13.081. JACI is the first in the field of Allergology and the sixth in the field of Immunology. We can conclude that the results of this project are very important for the novel diagnostic approaches and therapeutic concepts and that some diagnostic parts are already used in clinical settings.

With this project we accelerated the transfer of novel diagnostic platforms in clinical practice. With this transfer we improved the treatment and quality of life of Slovenian patients with allergic diseases. In this transfer we included platforms, related to the use of recombinant allergens and basophil activation test. A part of this project resulted in an European patent. We strongly believe that our patent will show positive economic effects both in Slovenia and in wider European area and further bust Slovenian research in the field of allergic diseases.