Primary airway epithelial cell culture provides a valuable tool for studying cell differentiation, cell–cell interactions, and the role of immune system factors in asthma pathogenesis. In this chapter, we discuss the application of mouse tracheal epithelial cell cultures for the st ...
The role of macrophage subsets in allergic diseases in vivo is under current investigation. These cells perform sentinel functions in the lung, the skin, and the gastrointestinal mucosa. Their interface with environmental cues influences the initiation, progression, developmen ...
Eosinophils are prominent in allergic diseases, and their effector functions are studied in numerous gene-deleted and transgenic mouse models. However, mouse eosinophils and human eosinophils are not structurally or functionally equivalent, and assays designed to evaluate t ...
Lymphocyte characterization is primarily based on the differential expression of surface markers. In this context, flow-cytometry analysis (FACS) is an exceptional technique that not only allows the identification of B-cell subsets, but can also be used to evaluate cell function, ac ...
T lymphocytes play positive and negative roles in the pathogenesis of allergic disease. Isolation and functional characterization of T lymphocyte subpopulations is an important aspect of understanding allergy models and allergy therapies. Measurement of the T cell surface prot ...
Antigen presenting cells (APCs) such as dendritic cells (DCs) and macrophages comprise a relatively small fraction of leukocytes residing in lymphoid and non-lymphoid tissues. Accordingly, functional analyses of these cells have been hampered by low cell yields. Also, alveolar mac ...
The creation of bone marrow and fetal liver chimeric mice has proven to be a valuable tool in the field of immunology. Chimeric mice are used to study the contribution of various cell types of hematopoietic versus non-hematopoietic origin in the course of an immune response. In this chapter, we descri ...
Multiparametric flow cytometry is a powerful technique that allows the quantification and characterization of heterogeneous populations of cells. Advances in flow cytometric instrumentation, software, and reagents are occurring at a rapid pace, and flow cytometric methods a ...
Transcript profiling reveals valuable insights to molecular and cellular activity related to disease. Gene expression profiles provide clues as to how tissues or cells in a particular environment may respond to stimuli. Gene-targeted examination of transcript changes is accomp ...
The increased incidence of asthma over the last 50 years in developed countries has been associated with a decrease in infections acquired early in childhood. These early infections are thought to shape subsequent immune responses. Although there have been multiple clinical associat ...
Isolated tracheal rings have often been used to directly measure the contractile output of airway smooth muscle (ASM). Here, we describe the method for excising murine tracheas, mounting tracheal rings in organ baths, and measuring the isometric forces generated by the ASM when stimulated ...
The generation of transgenic mouse models has been a powerful technique for several decades and is still widely used. There have been many manuals and general reviews of this technology. This chapter is designed to be a “how-to” resource with detailed specifics.
Airway remodeling is a term used to collectively indicate bronchial structural changes that may lead to irreversible airflow obstruction and progressive decline in lung function in asthmatic patients. Bronchial myofibroblasts contribute to airway remodeling by producing c ...
The bronchial epithelium has been increasingly recognized as an important immunomodulatory compartment in asthma and other lung diseases. Clara cells, which comprise the nonciliated secretory epithelial cells, are an important epithelial cell type with functions in the regul ...
This chapter describes the use of bilateral vagotomy as a tool for determining autonomic regulation of airway responses to the exogenous bronchoconstrictor thromboxane mimetic U46619 in an acute model of asthma in the mouse. Mice receive a sensitization of ovalbumin (OVA) and adjuvant f ...
This chapter provides an outline of the procedures necessary to measure airway hyperresponsiveness to inhaled methacholine in mouse models of allergic lung disease. We present a method for acquiring detailed measurements of respiratory mechanics using broadband low-frequen ...
Sensitization of mice to real-life allergens or harmless antigen with the use of adjuvants will lead to the induction of DAMPs in the immune system. We have shown that the Th2-inducing adjuvant aluminum hydroxide or exposure of the airways to house dust mite leads to the release of DAMPs: uric acid, ATP, a ...
The ability to accurately mimic normal processes for sensitization and allergen challenge in an experimental animal model are useful in that they allow researchers to critically manipulate the complex interactions of multiple cell types. In the context of the allergic lung, multiple c ...
Mouse models of allergic airway inflammation have proven essential in understanding the mechanisms and pathophysiology underling human asthma. There is a diverse range of mouse models described in the literature that typically vary slightly by allergen, duration of exposure, and r ...
We describe a method for allergic rhinitis (AR) induction in mice. Methodology involves nasal infusions of small volumes of ovalbumin for both initial sensitization and challenges. The latter are frequent and carried out over several weeks. This methodology more closely resembles nat ...
