TheDermatophagoides pteronyssinusGroup 2 Allergen Contains a Universally Immunogenic T Cell Epitope

Wu, Bo; Elst, Luc Vander; Carlier, Vincent; Jacquemin, Marc; Saint-Remy, Jean-Marie

doi:10.4049/jimmunol.169.5.2430

Cited by 15 publications

(19 citation statements)

References 33 publications

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“…The latter finding reconciles the results presented here with the X-ray crystallographic studies of Johannessen et al [14], who used ammonium sulphate precipitation. In the study by Wu et al [13], using undisclosed monoclonal antibodies to validate the structure of their rDer p 2 secreted from P. pastoris, the binding of these antibodies would be consistent with the observation here, i.e. even in the absence of the natural secondary structure, the IgE binding of rDer p 2 secreted from P. pastoris was largely intact.…”

Section: Discussionsupporting

confidence: 68%

“…Firstly, rDer p 2 was prepared by a single gel filtration step according to Wu et al [13]. Secondly, the allergen from this preparation was precipitated with 10% trichloro-acetic acid (TCA) and redissolved in PBS.…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, since Der p 2 and Der f 2 are secreted from P. pastoris [13,14], expression in this host could have the advantage of being more amenable to screening assays, e.g. for random site-directed mutagenesis studies.…”

Section: Introductionmentioning

confidence: 99%

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Structural and IgE Binding Analyses of Recombinant Der p 2 Expressed from the Hosts Escherichia coli and Pichia pastoris

Tanyaratsrisakul

Malainual²,

Jirapongsananuruk³

et al. 2009

Int Arch Allergy Immunol

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Background: The house dust mite allergen Der p 2 is one of the most important indoor allergens associated with allergic disease. Recombinant Der (rDer) p 2 with high IgE binding activity can be readily produced in Escherichia coli and Pichia pastoris, but the structure and IgE binding of the different methods of preparation have not been compared. Methods: Secondary structure was assessed by circular dichroism (CD). Intrinsic fluorescence and hydrophobic probe (1-anilinonaphthalene 8-sulphonic acid, ANS) were used to study the Der p 2 hydrophobic cavity. IgE binding was assessed by ELISA inhibition. Results: CD analysis showed the expected secondary structure for both nDer p 2 and refolded Der p 2 prepared from E. coli inclusion bodies but primarily random structure for Der p 2 secreted from P. pastoris. The secreted product, however, had disulphide bonding and could be refolded to a similar structure to natural Der (nDer) p 2 after precipitation with trichloro-acetic or ammonium sulphate. ANS binding and intrinsic Trp92 fluorescence showed that all recombinant proteins were different to nDer p 2 and that the allergen secreted from P. pastoris did not form a hydrophobic cavity. Despite the marked structural changes, all preparations of Der p 2 had similar IgE binding to nDer p 2. Conclusion: Despite almost identical IgE binding, rDer p 2 prepared from both E. coli and P. pastoris showed structural differences to nDer p 2. Der p 2 secreted from P. pastoris lacked most of the natural structure, but refolding could induce the natural structure.

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Section: Discussionsupporting

confidence: 68%

Section: Methodsmentioning

confidence: 99%

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Structural and IgE Binding Analyses of Recombinant Der p 2 Expressed from the Hosts Escherichia coli and Pichia pastoris

Tanyaratsrisakul

Malainual²,

Jirapongsananuruk³

et al. 2009

Int Arch Allergy Immunol

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“…p21-35 has been extensively studied in our laboratory: it contains both a major promiscuous T cell epitope and a dominant B epitope (17), thereby providing a suitable sequence for the study of tolerance induction at B and T cell levels. Injection of p21-35 in conventional adjuvant elicits strong specific Ab responses, while the same peptide under tolerogenic conditions results in prevention and partial suppression of the immune response toward Der p 2 (18).…”

Section: In Vivo Induction Of Type 1-like Regulatory T Cells Using Gementioning

confidence: 99%

In Vivo Induction of Type 1-Like Regulatory T Cells Using Genetically Modified B Cells Confers Long-Term IL-10-Dependent Antigen-Specific Unresponsiveness

Ahangarani

Janssens

VanderElst

et al. 2009

The Journal of Immunology

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“…We have previously demonstrated that p21 contained a promiscuous T cell epitope that was able to bind to MHC class II alleles of at least four different mouse haplotypes (33). Ile 28 in the p21 sequence was shown to act as a major MHC class II anchoring residue for each mouse strain.…”

Section: Wehi Cell Lysis Depends On Cognate Interaction With P21 Bounmentioning

confidence: 99%

CD4+CD25+ T Cells Lyse Antigen-Presenting B Cells by Fas-Fas Ligand Interaction in an Epitope-Specific Manner

Janssens

Carlier

et al. 2003

The Journal of Immunology

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128

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Suppression by regulatory T cells is now acknowledged to play a key role in the down-regulation of T cell responses to foreign and self Ags. In addition to the naturally occurring CD4+CD25+ population, several subtypes of induced regulatory cells have been reported, but their mechanisms of action remain unclear. Conversely, cytotoxic CD4+ cells that lyse cells presenting their cognate peptide have been described, but their potential role in immunoregulation remains to be delineated. A CD4+ T cell line derived from BALB/c mice immunized with peptide 21–35, containing a major T cell epitope of a common allergen, Dermatophagoides pteronyssinus group 2 allergen, was found to lyse the Ag-presenting WEHI cell line via Fas-Fas ligand and only in the presence of the cognate peptide. Cytolytic activity was likewise shown for other T cell lines and occurred even after a single cycle of in vitro stimulation. Moreover, T cells that efficiently lysed WEHI cells were unresponsive to stimulation with their cognate Ag and were dependent on IL-2 for growth and survival, which was reflected in a constitutive expression of CD25 independently of activation status. Proliferating B cells were also killed by the CTLs. By lysing Ag-presenting B cells in an epitope-specific manner, the nonproliferating CTLs were shown to down-regulate the proliferation of bystander T cells. These data demonstrate that cytotoxic CD4+CD25+ T cells that lack proliferation capacities have the potential to down-regulate an immune response by killing Ag-presenting B cells. This could represent an important and specific down-regulatory mechanism of secondary immune responses in vivo.

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TheDermatophagoides pteronyssinusGroup 2 Allergen Contains a Universally Immunogenic T Cell Epitope

Cited by 15 publications

References 33 publications

Structural and IgE Binding Analyses of Recombinant Der p 2 Expressed from the Hosts <i>Escherichia coli</i> and <i>Pichia pastoris</i>

Structural and IgE Binding Analyses of Recombinant Der p 2 Expressed from the Hosts <i>Escherichia coli</i> and <i>Pichia pastoris</i>

In Vivo Induction of Type 1-Like Regulatory T Cells Using Genetically Modified B Cells Confers Long-Term IL-10-Dependent Antigen-Specific Unresponsiveness

CD4+CD25+ T Cells Lyse Antigen-Presenting B Cells by Fas-Fas Ligand Interaction in an Epitope-Specific Manner

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