Abstract
Allergen-specific Immunoglobulin E (IgE) determination lies at the heart of diagnosis of sensitization to food and other allergens. In the past few years, reporter systems capable of detecting the presence of allergen-specific IgE have been developed by several labs. These rely on humanized rat basophil leukemia cell lines stably transfected with reporter genes such as firefly luciferase. In this chapter, we describe protocols for the use of the RS-ATL8 cell line (IgE cross-linking-induced luciferase expression; EXiLE) in 96-well and 384-well formats. We also describe optional treatment steps for enveloped virus and complement inactivation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Hill DJ, Heine RG, Hosking CS (2004) The diagnostic value of skin prick testing in children with food allergy. Pediatr Allergy Immunol 15:435–441
Kepley CL, Youssef L, Andrews RP et al (1999) Syk deficiency in nonreleaser basophils. J Allergy Clin Immunol 104:279–284
Küster H, von Manstein S, Ruocchio-Wiglinghaus S et al (2005) Mutations in the high-affinity IgE receptor beta-chain are not associated with nonresponder status. Allergy 60:1040–1045
Wan D, Wang X, Nakamura R et al (2014) Use of humanized rat basophil leukemia (RBL) reporter systems for detection of allergen-specific IgE sensitization in human serum. Methods Mol Biol 1192:177–184
Taudou G, Varin-Blank N, Jouin H et al (1993) Expression of the alpha chain of human Fc epsilon RI in transfected rat basophilic leukemia cells: functional activation after sensitization with human mite-specific IgE. Int Arch Allergy Immunol 100:344–350
Miller L, Blank U, Metzger H, Kinet JP (1989) Expression of high-affinity binding of human immunoglobulin E by transfected cells. Science 244:334–337
Wan D, Ludolf F, Alanine DGW et al (2014) Use of humanised rat basophilic leukaemia cell line RS-ATL8 for the assessment of allergenicity of Schistosoma mansoni proteins. PLoS Negl Trop Dis 8:e3124
Nakamura R, Ishiwatari A, Higuchi M et al (2012) Evaluation of the luciferase assay-based in vitro elicitation test for serum IgE. Allergol Int 61:431–437
Wang X, Cato P, Lin H-C et al (2013) Optimisation and use of humanised RBL NF-AT-GFP and NF-AT-DsRed reporter cell lines suitable for high-throughput scale detection of allergic sensitisation in array format and identification of the ECM-integrin interaction as critical factor. Mol Biotechnol 56:136–146
Nakamura R, Uchida Y, Higuchi M et al (2010) A convenient and sensitive allergy test: IgE crosslinking-induced luciferase expression in cultured mast cells. Allergy 65:1266–1273
Dibbern DA, Palmer GW, Williams PB et al (2003) RBL cells expressing human Fc epsilon RI are a sensitive tool for exploring functional IgE-allergen interactions: studies with sera from peanut-sensitive patients. J Immunol Methods 274:37–45
Poulsen LK, Sørensen TB (1993) Elimination of viral infection risk from blood samples for allergy testing. Allergy 48:207–208
Acknowledgments
E.A.A. was funded by a University of Nottingham Vice-Chancellor’s Scholarship for Research Excellence (International) and the Gordon Memorial College Trust Fund. Sera from milk allergic individuals and fractionated milk allergens were kindly provided by M. Alcocer, University of Nottingham. Our thanks to Colin Fitzsimmons (University of Cambridge), who introduced us to the use of the resin for detergent removal.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Ali, E.A., Nakamura, R., Falcone, F.H. (2017). Use of Humanized RS-ATL8 Reporter System for Detection of Allergen-Specific IgE Sensitization in Human Food Allergy. In: Lin, J., Alcocer, M. (eds) Food Allergens. Methods in Molecular Biology, vol 1592. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6925-8_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6925-8_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6923-4
Online ISBN: 978-1-4939-6925-8
eBook Packages: Springer Protocols