title: Subcutaneous and Sublingual Immunotherapy in a Mouse Model of Allergic Asthma.
abstract: Allergic asthma, caused by inhaled allergens such as house dust mite or grass pollen, is characterized by reversible airway obstruction, associated with an eosinophilic inflammation of the airways, as well as airway hyper responsiveness and remodeling. The inhaled allergens trigger a type-2 inflammatory response with involvement of innate lymphoid cells (ILC2) and Th2 cells, resulting in high production of immunoglobulin E (IgE) antibodies. Consequently, renewed allergen exposure results in a classic allergic response with a distinct early and late phase, both resulting in bronchoconstriction and shortness of breath. Allergen specific immunotherapy (AIT) is the only treatment that is capable of modifying the immunological process underlying allergic responses including allergic asthma and both subcutaneous AIT (SCIT) as well as sublingual AIT (SLIT) have proven clinical efficacy in long term suppression of the allergic response. Although these treatments are very successful for rhinitis, application of AIT in asthma is hampered by variable efficacy, long duration of treatment, and the risk of severe side-effects. A more profound understanding of the mechanisms by which AIT achieves tolerance to allergens in sensitized individuals is needed to improve its efficacy. Mouse models have been very valuable as a preclinical model to characterize the mechanisms of desensitization in AIT and to evaluate novel approaches for improved efficacy. Here, we present a rapid and reproducible mouse model for allergen-specific immunotherapy. In this model, mice are sensitized with two injections of allergen absorbed to aluminum hydroxide to induce allergic sensitization, followed by subcutaneous injections (SCIT) or sublingual administrations (SLIT) of the allergen as immunotherapy treatment. Finally, mice are challenged by three intranasal allergen administrations. We will describe the protocols as well as the most important read-out parameters including measurement of invasive lung function measurements, serum immunoglobulin levels, isolation of broncho-alveolar lavage fluid (BALF), and preparation of cytospins. Moreover, we describe how to restimulate lung single cell suspensions, perform flow cytometry measurements to identify populations of relevant immune cells, and perform ELISAs and Luminex assays to measure the cytokine concentrations in BALF and lung tissue.
authors.affiliation: Department of Pathology and Medical Biology, Laboratory of Experimental Pulmonology and Inflammation Research (EXPIRE), University Medical Center Groningen, University of Groningen, Hanzeplein 1 (EA52/T2.212), 9713 GZ, Groningen, The Netherlands.|Groningen Research Institute for Asthma and COPD (GRIAC), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
authors.forename: Laura
authors.initials: L
authors.lastname: Hesse
authors.affiliation: Department of Pathology and Medical Biology, Laboratory of Experimental Pulmonology and Inflammation Research (EXPIRE), University Medical Center Groningen, University of Groningen, Hanzeplein 1 (EA52/T2.212), 9713 GZ, Groningen, The Netherlands. M.C.Nawijn@umcg.nl.|Groningen Research Institute for Asthma and COPD (GRIAC), University Medical Center Groningen, University of Groningen, Groningen, The Netherlands. M.C.Nawijn@umcg.nl.
authors.forename: Martijn C
authors.initials: MC
authors.lastname: Nawijn
mesh_terms: D000485:Allergens / Q000008:administration & dosage*; D000818:Animals; D052179:Antigens, Plant / Q000008:administration & dosage*; D001249:Asthma / Q000139:chemically induced / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology / Q000276:immunology; D045424:Complex Mixtures / Q000008:administration & dosage; D016207:Cytokines / Q000096:biosynthesis; D004195:Disease Models, Animal; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D007108:Immune Tolerance / Q000187:drug effects; D007073:Immunoglobulin E / Q000096:biosynthesis; D007279:Injections, Subcutaneous / Q000379:methods*; D008168:Lung / Q000276:immunology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D010047:Ovalbumin / Q000008:administration & dosage; D010936:Plant Extracts / Q000008:administration & dosage*; D039981:Pyroglyphidae / Q000737:chemistry / Q000276:immunology; D063729:Sublingual Immunotherapy / Q000379:methods*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D045424:Complex Mixtures; D016207:Cytokines; D010936:Plant Extracts; C451253:grass pollen extract, alum-adsorbed; D007073:Immunoglobulin E; D010047:Ovalbumin
keywords: Allergic asthma; Bronchoalveolar lavage; Eosinophilia; FlexiVent; Flow cytometry; Grass pollen (GP); House dust mite (HDM); Mouse model; Subcutaneous immunotherapy (SCIT); Sublingual immunotherapy (SLIT)