Conformational IgE Epitope Mapping of Der p 2 and the Evaluations of Two Candidate Hypoallergens for Immunotherapy

Scientific Reports
Kavita Reginald, Fook Tim Chew

Abstract

Epitope mapping of Der p 2, a clinically important dust-mite allergen is the first step in designing immunotherapy hypoallergen vaccine candidates. Twenty-one single alanine mutants of Der p 2 were generated and their secondary structure was analysed using circular dichroism spectra. Only one mutant, K96A resulted in a misfolded protein. All mutants were tested for serum IgE reactivity using serum from dust mite allergic individuals by immuno dot-blots. Mutations to five residues, N10, E25, K77, K96 and E102 consistently showed reduced IgE reactions compared to wild-type Der p 2, and therefore these residues constitute the major IgE epitopes of Der p 2. Two mutants with consistent low IgE binding, K96A and E102A, were subsequently evaluated as hypoallergen candidates. IgG antibodies raised in mice against both mutants could inhibit human IgE-binding to WT Der p 2. Both mutants had intact T-cell epitopes as they were able to stimulate peripheral blood mononuclear cell proliferation similar to WT Der p 2. However, a switch in Th1:Th2 cytokine profile was not observed. In summary, we have identified the major conformational epitopes of Der p 2, and evaluated two Der p 2 hypoallergen vaccine candidates for immunotherapy.

References

Mar 1, 1990·Journal of Molecular Graphics·G Vriend
Oct 1, 1982·The Journal of Allergy and Clinical Immunology·G J GleichJ W Yunginger
May 1, 1997·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·A M Smith, M D Chapman
Mar 27, 1998·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·G A HakkaartR van Ree
Sep 2, 1998·The Journal of Allergy and Clinical Immunology·S R Durham, S J Till
Jul 31, 1999·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·T BallR Valenta
Aug 12, 1999·The New England Journal of Medicine·S R DurhamK T Nouri-Aria
Dec 14, 1999·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·W R Thomas, W Smith
Dec 22, 1999·Allergy·F T ChewB W Lee
Jun 11, 2002·Journal of Molecular Biology·U DerewendaD C Benjamin
Jul 3, 2002·Nature Reviews. Immunology·Rudolf Valenta
Feb 7, 2004·The Journal of Immunology : Official Journal of the American Association of Immunologists·Piia KarisolaHarri Alenius
Aug 18, 2004·Proceedings of the National Academy of Sciences of the United States of America·V NiederbergerR Valenta
Oct 8, 2004·The Journal of Immunology : Official Journal of the American Association of Immunologists·Jens HolmMichael D Spangfort
Apr 20, 2005·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·M I KidonA Balakrishnan
Apr 6, 2006·The Journal of Immunology : Official Journal of the American Association of Immunologists·Siew Leong ChanYu Keung Mok
Jul 26, 2006·International Archives of Allergy and Immunology·R J J van NeervenP A Würtzen
May 31, 2008·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·P A WürtzenH Ipsen
Mar 3, 2010·Annual Review of Immunology·Rudolf ValentaSusanne Vrtala
Jul 14, 2012·The Journal of Allergy and Clinical Immunology·Kuan-Wei ChenSusanne Vrtala
May 27, 2014·Current Treatment Options in Allergy·Katharina MarthVerena Niederberger

❮ Previous
Next ❯

Citations

Sep 27, 2019·The Journal of Immunology : Official Journal of the American Association of Immunologists·Jill GlesnerAnna Pomés
Jan 9, 2020·Expert Opinion on Therapeutic Patents·Eduardo Santos da SilvaNeuza Maria Alcantara-Neves
Aug 18, 2020·Molecular Biology Reports·Hui Cao, Zhigang Liu
Aug 24, 2019·Scientific Reports·Kavita ReginaldFook Tim Chew
Nov 28, 2018·Allergy, Asthma & Immunology Research·Bhakkawarat KulwanichSurapon Piboonpocanun
Mar 22, 2019·Scientific Reports·Sze Lei PangChyan Leong Ng
Sep 29, 2020·Frontiers in Immunology·Anna PomésMaksymilian Chruszcz
Sep 11, 2020·The Journal of Immunology : Official Journal of the American Association of Immunologists·Geoffrey A MuellerAnna Pomés
Dec 2, 2020·PloS One·Véronique GomordLouis-P Vézina
Jan 15, 2021·Scientific Reports·Sze Lei PangFook Tim Chew
Apr 19, 2021·Allergy·Maksymilian ChruszczHeimo Breiteneder
Sep 1, 2021·The Journal of Immunology : Official Journal of the American Association of Immunologists·Min Hwa HongIn Sik Kim

❮ Previous
Next ❯

Methods Mentioned

BETA
circular dichroism
dot-blots
dot blot
dot blots
ELISA
ELISAs

Software Mentioned

Microimage

Related Concepts

Related Feeds

Allergy and Asthma

Allergy and asthma are inflammatory disorders that are triggered by the activation of an allergen-specific regulatory t cell. These t cells become activated when allergens are recognized by allergen-presenting cells. Here is the latest research on allergy and asthma.