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Chapter 83 : Assay Methods for Measurement of Mediators and Markers of Allergic Inflammation

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Assay Methods for Measurement of Mediators and Markers of Allergic Inflammation, Page 1 of 2

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Abstract:

Assays facilitating the measurement of mediators and markers of allergic inflammation have evolved during the past 40 years, arising from two primary discoveries: first, that immunoglobulin E (IgE) is the source of reaginic activity in serum, and second, that tissue mast cells and blood basophils—cells expressing the high-affinity receptor for IgE (FcεR1)—respond to allergen by releasing the most relevant mediators underlying immediate hypersensitivity reactions. Naturally, skin testing has long been used to assess the mast cell response to allergen. Basophils, however, by virtue of their accessibility and the fact that they are the sole source of many of these mediators among blood leukocytes, continue to dominate the performance of assays assessing IgE-mediated responses.

Citation: Schroeder J, Peebles R, Guerrerio P. 2016. Assay Methods for Measurement of Mediators and Markers of Allergic Inflammation, p 801-814. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch83
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FIGURE 1

Diagrammatic representation of preparation of peripheral blood basophils by discontinuous Percoll density centrifugation. Leukocyte interface cells (buffy coat) are prepared from whole blood by centrifugation. Diluted buffy-coat cell suspensions are layered onto buffered isotonic gradients consisting of two Percoll densities (d) of 1.072 and 1.082 g/ml. Following centrifugation (700 × , 20 min, at 24°C), basophils band on the 1.082-g/ml Percoll interface and range from 5 to 50% purity. v/v, vol/vol.

Citation: Schroeder J, Peebles R, Guerrerio P. 2016. Assay Methods for Measurement of Mediators and Markers of Allergic Inflammation, p 801-814. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch83
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