| Literature DB >> 28063042 |
Eline Haspeslagh1,2, Nincy Debeuf1,3, Hamida Hammad1,3, Bart N Lambrecht4,5.
Abstract
Allergic asthma is a heterogeneous inflammatory lung disease affecting millions of people worldwide and with a steadily increasing incidence. Mouse models have been of utmost importance in uncovering key inflammatory cell types, cytokines, and pathways in the development and maintenance of allergic asthma. Historically, the mainstay in experimental asthma research was sensitizing rodents to the model protein antigen ovalbumin (OVA) with the pro-Th2 adjuvant aluminum hydroxide, followed by repetitive OVA exposures to the airways to initiate a Th2-skewed adaptive immune response leading to eosinophilic airway inflammation and airway hyperreactivity (AHR). In the last 5 years, OVA is often replaced by naturally occurring allergens such as house dust mite (HDM) or cockroach extracts, but the principle of first sensitizing and then repetitively challenging mice with the same antigen is unchanged. Here, we describe an often used and relevant HDM-based protocol to establish acute allergic asthma, and the methods we have developed to rapidly analyze inflammatory cell infiltration in the bronchalveolar lavage fluid by flow cytometry. Moreover, we explain the methods to restimulate T cells from lung-draining mediastinal lymph nodes with HDM to allow the measurement of cytokine secretion profiles of allergen reactive T cells.Entities:
Keywords: Acute allergic asthma; Bronchoalveolar lavage; Eosinophilia; Flow cytometry; House dust mite (HDM); Mediastinal lymph nodes; Murine model; Ovalbumin (OVA)
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Year: 2017 PMID: 28063042 DOI: 10.1007/978-1-4939-6786-5_10
Source DB: PubMed Journal: Methods Mol Biol ISSN: 1064-3745