Abstract
Soluble cytokine receptors have proven to be very useful biomarkers in a large variety of diseases, including cancer, infections, and chronic inflammatory diseases. These soluble receptors are produced by proteolytic cleavage or alternative splicing. Several cytokine receptors including tumor necrosis factor receptor 2 (TNFR2) can be generated by both mechanisms. However, the conventional ELISA systems do not differentiate between these two types of soluble receptors. We describe a sandwich ELISA to specifically quantify soluble TNFR2 protein generated by alternative splicing. This method requires the use of a capturing monoclonal antibody (mAb) specific of an epitope present in the soluble TNFR2 generated by alternatively splicing but absent in the proteolytically generated isoform. Here we present a detailed protocol for the production and validation of such a mAb. This method has the potential to be applied for measuring other soluble cell surface molecules generated by alternative splicing.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Rose-John S, Heinrich PC (1994) Soluble receptors for cytokines and growth factors: generation and biological function. Biochem J 300:281ā290
Heaney ML, Golde DW (1996) Soluble cytokine receptors. Blood 87:847ā857
Levine SJ (2008) Molecular mechanisms of soluble cytokine receptor generation. J Biol Chem 283:14177ā14181
Heaney ML, Golde DW (1998) Soluble receptors in human disease. J Leukoc Biol 64:135ā146
Ernandez T, Mayadas TN (2009) Immunoregulatory role of TNFalpha in inflammatory kidney diseases. Kidney Int 76:262ā276
Heemann C, Kreuz M, Stoller I, Schoof N, von Bonin F, Ziepert M et al (2012) Circulating levels of TNF receptor II are prognostic for patients with peripheral T-cell non-Hodgkin lymphoma. Clin Cancer Res 18:3637ā3647
Hivert MF, Sullivan LM, Shrader P, Fox CS, Nathan DM, DāAgostino RB Sr et al (2010) The association of tumor necrosis factor alpha receptor 2 and tumor necrosis factor alpha with insulin resistance and the influence of adipose tissue biomarkers in humans. Metabolism 59:540ā546
Gohda T, Niewczas MA, Ficociello LH, Walker WH, Skupien J, Rosetti F et al (2012) Circulating TNF receptors 1 and 2 predict stage 3 CKD in type 1 diabetes. J Am Soc Nephrol 23:516ā524
Aderka D, Wysenbeek A, Engelmann H, Cope AP, Brennan F, Molad Y et al (1993) Correlation between serum levels of soluble tumor necrosis factor receptor and disease activity in systemic lupus erythematosus. Arthritis Rheum 36:1111ā1120
Gabay C, Cakir N, Moral F, Roux-Lombard P, Meyer O, Dayer JM et al (1997) Circulating levels of tumor necrosis factor soluble receptors in systemic lupus erythematosus are significantly higher than in other rheumatic diseases and correlate with disease activity. J Rheumatol 124:303ā308
Schrƶder J, StĆ¼ber F, Gallati H, Schade FU, Kremer B (1995) Pattern of soluble TNF receptors I and II in sepsis. Infection 23:143ā148
Lainez B, Fernandez-Real JM, Romero X, Esplugues E, CaƱete JD, Ricart W et al (2004) Identification and characterization of a novel spliced variant that encodes human soluble tumor necrosis factor receptor 2. Int Immunol 16:169ā177
CaƱete JD, Albaladejo C, HernĆ”ndez MV, LaĆnez B, Pinto JA, RamĆrez J et al (2012) Clinical significance of high levels of soluble tumour necrosis factor-Ī± receptor-2 produced by alternative splicing in rheumatoid arthritis: a longitudinal prospective cohort study. Rheumatology (Oxford) 50:721ā728
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Romero, X., CaƱete, J.D., Engel, P. (2014). Determination of Soluble Tumor Necrosis Factor Receptor 2 Produced by Alternative Splicing. In: Bayry, J. (eds) The TNF Superfamily. Methods in Molecular Biology, vol 1155. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0669-7_16
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0669-7_16
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0668-0
Online ISBN: 978-1-4939-0669-7
eBook Packages: Springer Protocols