ahmed-identificationand-2015.pdf (1.06 MB)
Identification and characterization of a novel IL-4 receptor α chain (IL-4Rα) antagonist to inhibit IL-4 signalling
journal contribution
posted on 2015-01-01, 00:00 authored by Nayyar Ahmed, Pathum Dhanapala, Cenk SuphiogluCenk SuphiogluBACKGROUND/AIMS: In recent times, allergy has become a financial, physical and psychological burden to the society as a whole. In allergic cascades, cytokine IL-4 binds to IL-4 receptor (IL-4R), consequently producing allergen-specific IgE antibodies by B cells. In addition, among other functions, IL-4 is also responsible for B and T cell proliferation and differentiation. Hence, characterization of novel antagonists that inhibit IL-4 signalling forms the overall aim of this study. METHODS: Phage display was used to screen a random 12-mer synthetic peptide library with a human IL-4Rα to identify peptide candidates. Once identified, the peptides were commercially synthesized and used for in vitro immunoassays. RESULTS: We have successfully used phage display to identify M13 phage clones that demonstrated specific binding to IL-4Rα. The peptide N1 was synthesized for use in ELISA, demonstrating significant binding to IL-4Rα and inhibiting interaction with cytokine IL-4. Furthermore, the peptide was tested in a transfected HEK-Blue IL-4 reporter cell line model, which produces alkaline phosphatase (AP). QUANTI-Blue, a substrate, breaks down in the presence of AP producing a blue coloration. Using this colorimetric analysis, >50% inhibition of IL-4 signalling was achieved. CONCLUSION: We have successfully identified and characterised a synthetic peptide antagonist against IL-4Rα, which effectively inhibits IL-4 interaction with the IL-4Rα in vitro. Since IL-4 interaction with IL-4Rα is a common pathway for many allergies, a prophylactic treatment can be devised by inhibiting this interaction for future treatment of allergies.
History
Journal
Cellular physiology and biochemistryVolume
36Issue
3Pagination
831 - 842Publisher
KargerLocation
Basal, SwitzerlandPublisher DOI
Link to full text
eISSN
1421-9778Language
engPublication classification
C Journal article; C1 Refereed article in a scholarly journalCopyright notice
2015, KargerUsage metrics
Categories
No categories selectedKeywords
Alkaline PhosphataseAnti-Allergic AgentsBiological AssayCell LineColorimetryGene ExpressionHEK293 CellsHumansInterleukin-4Interleukin-4 Receptor alpha SubunitPeptide LibraryPeptidesProtein BindingProtein IsoformsSignal TransductionScience & TechnologyLife Sciences & BiomedicineCell BiologyPhysiologyAllergyPhage displayIL-4 signalling pathwayIL-4 receptorIgEELISAHEK-Blue cell lineTUMOR-NECROSIS-FACTORPHAGE-DISPLAYALLERGIC DISEASESMONOCLONAL-ANTIBODIESFRAGMENT LIBRARIESPEPTIDE LIBRARIESSOLUBLE RECEPTORSFOOD ALLERGYPROTEIN
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC