In recent years, allergy research has focused on the causes and mechanisms of allergy. In parallel, there is also an impetus to try to understand mechanisms of natural tolerance and immunotherapy where allergy is being dampened. In Allergy: Methods and Protocols a groundbreaking new title from the Methods in Molecular Medicine series, leaders in the field provide assistance for researchers to gain insight into the molecular mechanisms involved in allergy by featuring an array of protocols. These cover a range of disciplines including allergy, immunology, cell biology and histology and include methods to investigate the cellular response to allergens, cytokine profile, MHC restriction, T regulatory cells. Techniques discussed include; B and T cell epitope mapping, characterization of allergens, conjugation of haptens, preparation of monoclonal antibodies, collection and sampling of airborne allergens, IgG antibodies and facilitated antigen blocking assays, identification and purification of mast cells and in situ hybridisation. Allergy: Methods and Protocols will be a remarkably useful bench tool for anyone embarking in or continuing with their research in allergy.
Written for:
Allergists, immunologists, immunotherapists, cell biologists, histologists, molecular biologists
Table of contents
List of chapters
1. Understanding the molecular mechanisms of allergy - Meinir Jones
2. T cell - primary culture from peripheral blood - Monika
Raulf-Heimsoth
3. Production of T cell lines - Helga Kahlert
4. Production of human T cell clones - Adrienne Verhoef
5. Mapping of human T-cell epitopes of allergens - Thomas Zeiler and
Tuomas Virtanen
6. Determining MHC restriction of T cell response - Mark Larche
7. Short term culture of CD8 cells and intracellular cytokine staining
- Beejal Vyas and Alistair Noble
8. Isolation, flow cytometric analysis and suppression assay of
CD4+CD25+ T regulatory cells - Hayley Jeal
9. Monocyte-derived dendritic cells as antigen presenting cells in T
cell proliferation and cytokine production - Sun-sang J Sung
10. Ultrasensitive ELISA for measurement of human cytokine responses in
primary culture - William P. Stefura, J. Darren Campbell, Renée
Douville, Monique J. Stinson, F. Estelle Simons, Allan B. Becker, and
Kent T. HayGlass
11. Quantification of human chemokine production in TLR-stimulated and
antigen-specific recall responses - Monique Stinson, Renee Douville,
Yuriy Lissitsyn, Melanie Blanchard, William Stefura, Estelle Simons,
Allan Becker, Peter Nickerson, Kevin Coombs, Kent HayGlass
12. Standardisation of allergen extracts - Jørgen Nedergaard Larsen and
Sten Dreborg
13. Immuno-electrophoresis for the characterisation of allergen
extracts - Gitte Nordskov Hansen and Jørgen Nedergaard Larsen
14. Conjugation of haptens - Ranulfo Lemus and Meryl H. Karol
15. Monoclonal antibodies - Helga Kahlert and Oliver Cromwell
16. Purification of antibodies - Per H Larsson and Anne Renström
17. Collection of air samples to quantify exposure to airborne
allergens - Susan Gordon
18. Assay of air sample eluates - Anne Renström and Susan Gordon
19. The halogen assay - a new technique for measuring airborne allergen
- Euan Tovey, Sandra De Lucca, Leanne Poulos and Tim O'Meara
20. Measurement of specific IgG anti-Fel d 1 antibodies - Meinir Jones
21. The facilitated-antigen binding (FAB) assay - a protocol to measure
allergen-specific inhibitory activity - James N Francis
22. Microscopic identification and purity determination of pollen
grains - Magdalena Rahl
23. Biopanning for the characterisation of allergen mimotopes -
Isabella Schöll and Erika Jensen-Jarolim
24. Identification of mast cells and mast cell subpopulations - Mark
Buckley and Andrew F Walls
25. Purification and characterisation of mast cell tryptase and chymase
- Alan R McEuen and Andrew F Walls
26. Experimental activation of mast cells and their pharmacological
modulation - Shaoheng He and Andrew F Walls
27. In situ hybridisation - Kayhan T. Nouri-Aria