PMID- 33226602 OWN - NLM STAT- MEDLINE DCOM- 20210326 LR - 20210326 IS - 1940-6029 (Electronic) IS - 1064-3745 (Linking) VI - 2223 DP - 2021 TI - Methods for Experimental Allergen Immunotherapy: Subcutaneous and Sublingual Desensitization in Mouse Models of Allergic Asthma. PG - 295-335 LID - 10.1007/978-1-0716-1001-5_20 [doi] AB - Allergic asthma is characterized by airway hyperresponsiveness, remodeling, and reversible airway obstruction. This is associated with an eosinophilic inflammation of the airways, caused by inhaled allergens such as house dust mite or grass pollen. The inhaled allergens trigger a type-2 inflammatory response with the involvement of innate lymphoid cells (ILC2) and Th2 cells, resulting in high immunoglobulin E (IgE) antibody production by B cells and mucus production by airway epithelial cells. As a consequence of the IgE production, subsequent allergen reexposure results in a classic allergic response with distinct early and late phases, 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. Both subcutaneous AIT (SCIT) as well as sublingual AIT (SLIT) have shown clinical efficacy in long-term suppression of the allergic response. Although AIT treatments are very successful for rhinitis, application in asthma is hampered by variable efficacy, long duration of treatment, and risk of severe side effects. A more profound understanding of the mechanisms by which AIT induces tolerance to allergens in sensitized individuals is needed to be able to improve its efficacy. Mouse models have been very valuable in preclinical research for characterizing the mechanisms of desensitization in AIT and evaluating novel approaches to improve its 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 adsorbed to aluminum hydroxide, followed by subcutaneous injections (SCIT) or sublingual administrations (SLIT) of allergen extracts as an immunotherapy treatment. Finally, mice are challenged by intranasal allergen administrations. We will also describe the protocols as well as the most important readout parameters for the measurements of invasive lung function, serum immunoglobulin levels, isolation of bronchoalveolar lavage fluid (BALF), and preparation of cytospin slides. Moreover, we describe how to perform ex vivo restimulation of lung single-cell suspensions with allergens, flow cytometry for identification of relevant immune cell populations, and ELISAs and Luminex assays for assessment of the cytokine concentrations in BALF and lung tissue. FAU - Hesse, Laura AU - Hesse L AD - Department of Pathology and Medical Biology, Laboratory of Experimental Pulmonology and Inflammation Research (EXPIRE), University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. AD - Groningen Research Institute for Asthma and COPD (GRIAC), University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. FAU - Petersen, Arjen H AU - Petersen AH AD - Department of Pathology and Medical Biology, Laboratory of Experimental Pulmonology and Inflammation Research (EXPIRE), University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. AD - Groningen Research Institute for Asthma and COPD (GRIAC), University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. FAU - Nawijn, Martijn C AU - Nawijn MC AD - Department of Pathology and Medical Biology, Laboratory of Experimental Pulmonology and Inflammation Research (EXPIRE), University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. m.c.nawijn@umcg.nl. AD - Groningen Research Institute for Asthma and COPD (GRIAC), University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. m.c.nawijn@umcg.nl. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - United States TA - Methods Mol Biol JT - Methods in molecular biology (Clifton, N.J.) JID - 9214969 RN - 0 (Adjuvants, Immunologic) RN - 0 (Allergens) RN - 0 (Complex Mixtures) RN - 0 (Cytokines) RN - 37341-29-0 (Immunoglobulin E) RN - 5QB0T2IUN0 (Aluminum Hydroxide) SB - IM MH - Adjuvants, Immunologic/administration & dosage MH - Administration, Intranasal MH - Allergens/*administration & dosage/immunology MH - Aluminum Hydroxide/administration & dosage MH - Animals MH - Asthma/immunology/pathology/*therapy MH - Bronchoalveolar Lavage Fluid/chemistry/cytology/immunology MH - Complex Mixtures/administration & dosage/immunology MH - Cytokines/genetics/immunology MH - *Disease Models, Animal MH - Ear MH - Eosinophils/immunology/pathology MH - Female MH - Humans MH - Immunoglobulin E/genetics/immunology MH - Injections, Subcutaneous MH - Lung/drug effects/immunology/pathology MH - Mice MH - Mice, Inbred BALB C MH - Mice, Inbred C57BL MH - Neutrophils/immunology/pathology MH - Pollen/chemistry/*immunology MH - Pyroglyphidae/chemistry/*immunology MH - Single-Cell Analysis/methods MH - Sublingual Immunotherapy/*methods OTO - NOTNLM OT - Allergic asthma OT - BALB/cByJ and C57BL/6 mouse models OT - Bronchoalveolar lavage OT - Eosinophilia OT - FlexiVent OT - Flow cytometry OT - Grass pollen (GP) OT - House dust mite (HDM) OT - Subcutaneous immunotherapy (SCIT) OT - Sublingual immunotherapy (SLIT) EDAT- 2020/11/24 06:00 MHDA- 2021/03/27 06:00 CRDT- 2020/11/23 12:12 PHST- 2020/11/23 12:12 [entrez] PHST- 2020/11/24 06:00 [pubmed] PHST- 2021/03/27 06:00 [medline] AID - 10.1007/978-1-0716-1001-5_20 [doi] PST - ppublish SO - Methods Mol Biol. 2021;2223:295-335. doi: 10.1007/978-1-0716-1001-5_20.