title: Conversion of Der p 23, a new major house dust mite allergen, into a hypoallergenic vaccine.
abstract: Der p 23, a new, major house dust mite (HDM) allergen that is recognized by >70% of HDM-allergic patients, has high allergenic activity and, therefore, must be considered an important component for HDM-specific immunotherapy. We constructed and characterized a hypoallergenic Der p 23 vaccine for HDM immunotherapy. Three nonallergenic peptides from the C-terminal IgE epitope-containing part of Der p 23 (P4, P5) and P6, a mutant peptide containing serines instead of cysteines, were identified. Peptides were fused to the hepatitis B virus-derived PreS domain as recombinant fusion proteins (i.e., PreS-2XP4P5 and PreS-4XP6) that were expressed in Escherichia coli and purified to homogeneity. Compared with Der p 23, PreS-2XP4P5 and PreS-4XP6 showed no relevant IgE reactivity and exhibited considerably reduced allergenic activity in basophil activation tests using blood from HDM-allergic patients. Upon immunization of rabbits, only PreS-2XP4P5 induced high levels of Der p 23-specific IgG Abs that inhibited binding of patients' IgE to Der p 23, comparable to IgG Abs induced with Der p 23, whereas Abs induced with PreS-4XP6 had only low blocking capacity. Additionally, IgG Abs induced with PreS-2XP4P5 inhibited Der p 23-induced basophil activation comparable to IgG Abs induced with Der p 23. Compared with Der p 23, PreS-2XP4P5 induced lower T cell proliferation but higher levels of the tolerogenic cytokine IL-10 and the Th1 cytokine IFN-γ in PBMCs from HDM-allergic patients, indicating an immunomodulatory capacity of the fusion protein. Therefore, PreS-2XP4P5 represents a promising candidate for immunotherapy of HDM-allergic patients.
authors.affiliation: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology, and Immunology, Medical University of Vienna, 1090 Vienna, Austria;
authors.forename: Srinita
authors.initials: S
authors.lastname: Banerjee
authors.forename: Milena
authors.initials: M
authors.lastname: Weber
authors.forename: Katharina
authors.initials: K
authors.lastname: Blatt
authors.forename: Ines
authors.initials: I
authors.lastname: Swoboda
authors.forename: Margit
authors.initials: M
authors.lastname: Focke-Tejkl
authors.forename: Peter
authors.initials: P
authors.lastname: Valent
authors.forename: Rudolf
authors.initials: R
authors.lastname: Valenta
authors.forename: Susanne
authors.initials: S
authors.lastname: Vrtala
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D001491:Basophils / Q000276:immunology* / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D016753:Interleukin-10 / Q000276:immunology; D008297:Male; D039981:Pyroglyphidae / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D018417:Th1 Cells / Q000276:immunology; D014612:Vaccines / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D000485:Allergens; D039741:Antigens, Dermatophagoides; C573415:IFNG protein, human; C508609:IL10 protein, human; D011994:Recombinant Proteins; D014612:Vaccines; D016753:Interleukin-10; D007073:Immunoglobulin E; D007371:Interferon-gamma
references.citation: J Allergy Clin Immunol. 2000 Feb;105(2 Pt 2):S503-8
references.pmid: 10669532
references.citation: Curr Top Microbiol Immunol. 2011;352:55-69
references.pmid: 21400223
references.citation: Allergy. 2012 May;67(5):609-21
references.pmid: 22339348
references.citation: Clin Exp Allergy. 2009 Jul;39(7):1088-98
references.pmid: 19400901
references.citation: J Allergy Clin Immunol. 2011 Apr;127(4):865-72
references.pmid: 21377719
references.citation: Methods. 2014 Mar 1;66(1):67-74
references.pmid: 23911838
references.citation: Clin Exp Allergy. 2000 Oct;30(10):1423-9
references.pmid: 10998019
references.citation: J Allergy Clin Immunol. 2009 May;123(5):1150-6
references.pmid: 19152967
references.citation: Mol Immunol. 2008 May;45(9):2486-98
references.pmid: 18295887
references.citation: J Immunol. 2004 May 1;172(9):5684-92
references.pmid: 15100313
references.citation: Allergy. 2011 Jun;66(6):775-83
references.pmid: 21352238
references.citation: Vaccine. 2006 Apr 5;24(15):2781-9
references.pmid: 16455169
references.citation: J Allergy Clin Immunol. 2010 Nov;126(5):1024-31, 1031.e1-8
references.pmid: 20638112
references.citation: Vaccine. 2011 Mar 3;29(11):2140-8
references.pmid: 21215346
references.citation: Ann Allergy Asthma Immunol. 2000 Oct;85(4):317-21
references.pmid: 11061476
references.citation: Curr Top Microbiol Immunol. 2011;352:85-105
references.pmid: 21562971
references.citation: Curr Top Microbiol Immunol. 2011;352:107-19
references.pmid: 21567311
references.citation: Clin Exp Allergy. 2011 Oct;41(10):1468-77
references.pmid: 21711470
references.citation: J Immunol. 2013 Apr 1;190(7):3068-78
references.pmid: 23440415
references.citation: Allergy. 2010 Feb;65(2):184-90
references.pmid: 19796217
references.citation: Clin Exp Allergy. 2012 Jun;42(6):966-75
references.pmid: 22909168
references.citation: Nature. 1981 Feb 12;289(5798):592-3
references.pmid: 7464922
references.citation: AAPS J. 2010 Sep;12(3):330-7
references.pmid: 20419360
references.citation: Eur J Clin Invest. 2008 Dec;38(12):959-65
references.pmid: 19021722
references.citation: Allergy Proc. 1994 Nov-Dec;15(6):329-33
references.pmid: 7721083
references.citation: Clin Exp Allergy. 2004 Apr;34(4):597-603
references.pmid: 15080813
references.citation: Int Arch Allergy Immunol. 2012;159(3):253-62
references.pmid: 22722650
references.citation: J Immunol. 2013 Apr 1;190(7):3059-67
references.pmid: 23460742
references.citation: Curr Top Microbiol Immunol. 2011;352:71-84
references.pmid: 21725898
references.citation: J Allergy Clin Immunol. 2001 Jan;107(1):87-93
references.pmid: 11149996
references.citation: FASEB J. 2001 Sep;15(11):2042-4
references.pmid: 11511525
references.citation: J Allergy Clin Immunol. 2011 Jun;127(6):1562-70.e6
references.pmid: 21411130
references.citation: Allergy. 2005 Oct;60(10):1287-300
references.pmid: 16134996
references.citation: Int Arch Allergy Immunol. 2011;155(2):95-105
references.pmid: 21196753
references.citation: J Allergy Clin Immunol. 2011 Jul;128(1):178-184.e7
references.pmid: 21513971
references.citation: Clin Exp Allergy. 1994 Aug;24(8):737-42
references.pmid: 7982123
references.citation: J Immunol. 2002 May 1;168(9):4576-84
references.pmid: 11971005
references.citation: Adv Virus Res. 1988;34:65-142
references.pmid: 3046258
references.citation: J Allergy Clin Immunol. 2002 Jul;110(1):102-9
references.pmid: 12110828
references.citation: J Allergy Clin Immunol. 2012 Aug;130(2):435-43.e4
references.pmid: 22789398
references.citation: Curr Opin Allergy Clin Immunol. 2012 Oct;12(5):555-63
references.pmid: 22885888
references.citation: Anal Biochem. 1986 May 15;155(1):83-8
references.pmid: 3454661