Characterization of Major Allergens of &lt;i&gt;Parietaria officinalis&lt;/i&gt;

Kahlert, Helga; Weber, Bernhard H. F.; Teppke, Manfred; Wahl, R.; Cromwell, Oliver; Fiebig, Helmut

doi:10.1159/000237213

Cited by 15 publications

(9 citation statements)

References 16 publications

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“…Bet v 1-specific mouse mAb of the IgG1 isotype (1D6, 1G3, 1G7, 2E10, 3B4, 4B10, 6B6, 6D10, 7G6, 9B11, 9D6, 10D6) were raised with birch pollen extract according to procedures described elsewhere [41, 42]. …”

Section: Methodsmentioning

confidence: 99%

Characterization of a Hypoallergenic Recombinant Bet v 1 Variant as a Candidate for Allergen-Specific Immunotherapy

Kahlert¹,

Suck²,

Weber³

et al. 2007

Int Arch Allergy Immunol

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Background: Recombinant allergens and especially their hypoallergenic variants are promising candidates for a more effective and safer specific immunotherapy. Methods: Physicochemical and immunological characteristics of a folding variant of recombinant Bet v 1 (rBet v 1-FV) were investigated in comparison to natural Bet v 1 (nBet v 1) and the correctly folded recombinant Bet v 1 (rBet v 1-WT) by SDS-PAGE, size exclusion chromatography, multi-angle light scattering, circular dichroism, immunoblotting and enzyme allergosorbent test inhibition assay for detection of IgE reactivity and ELISA with Bet v 1-specific monoclonal antibodies. The functional IgE reactivity of the different Bet v 1 proteins was investigated using basophil activation in terms of CD203c expression and histamine release. T cell reactivity was investigated using T cell lines raised from birch pollen-allergic subjects against nBet v 1. Immunogenicity was investigated in mice. Results: Physicochemical characterization revealed purity, homogeneity and monomeric properties of rBet v 1-FV. Unlike nBet v 1 and rBet v 1-WT, rBet v 1-FV showed almost no IgE binding in immunoblots. The reduction of allergenicity was further proved by IgE-binding inhibition assays, basophil activation and histamine release. T cell reactivity was completely conserved, as demonstrated by proliferation of Bet v 1-specific T cell lines with multiple epitope specificities. rBet v 1-FV showed strong immunogenicity in mice. Conclusions: Due to its reduced IgE reactivity and decreased capacity to activate basophils, but retained T cell reactivity and strong immunogenicity, rBet v 1-FV proved to be a very promising candidate for specific immunotherapy in birch pollen-allergic subjects.

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Section: Methodsmentioning

confidence: 99%

Characterization of a Hypoallergenic Recombinant Bet v 1 Variant as a Candidate for Allergen-Specific Immunotherapy

Kahlert¹,

Suck²,

Weber³

et al. 2007

Int Arch Allergy Immunol

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“…Purified major allergen Phl p 5 was prepared by affinity chromatography using the monoclonal antibody 1D11 [25, 26]coupled to protein G Sepharose 4 fast flow (Pharmacia, Freiburg, Germany) by a method described elsewhere [27]. …”

Section: Methodsmentioning

confidence: 99%

Reactivity of T Cells with Grass Pollen Allergen Extract and Allergoid

Kähler¹,

Stüwe²,

Cromwell³

et al. 1999

Int Arch Allergy Immunol

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Background: Successful allergen–specific immunotherapy is achieved with progressively increasing doses of allergen or allergoid. In order to gain further insight into the mechanism of action of allergoids several in vitro investigations were conducted. Methods: Peripheral blood mononuclear cells (PBMC) from grass pollen allergic and nonallergic subjects were stimulated with either grass pollen extract or allergoid and the proliferation and cytokine production (IL–5, IFN–γ) were measured. Similar investigations were performed with Phl p 5–specific T cell lines (TCL) and clones (TCC). Dendritic cells and PBMC were compared in terms of their relative efficacies as antigen–presenting cells. Results: Both allergen and allergoid induced proliferation and Th2 and Th1 cytokine synthesis by PBMC of allergic subjects, whereas PBMC of nonallergic subjects did not produce IL–5. The maximum level of IL–5 was obtained with a lower concentration than was necessary for maximal IFN–γ production. Higher stimulation doses of allergen and allergoid shifted the cytokine profiles towards a Th1 phenotype. TCL and TCC clearly showed reactivity with both allergen and allergoid when using autologous PBMC for antigen presentation, but compared with the native allergen the reactivity of the allergoid was reduced with most of the TCC. Using dendritic cells for antigen presentation a pronounced increase of stimulation of the TCC especially for the allergoids becomes obvious. Conclusion: In common with grass pollen allergen the corresponding allergoids possess a strong allergen–specific T cell–stimulating capacity. However, the degree of T cell stimulation by the allergoid seems to be dependent on the type of the antigen–presenting cell. Both, allergen and allergoid, can modulate T cell responses in a dose–dependent manner.

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“…Isotype controls were IgG1 (107.3), IgG2a (G155-178), and IgG2b (27-35) obtained from PharMingen. For secondary isotype control we used a mouse mAb 2F9 of the IgG1 isotype with specificity for Par o 1 (43) and FITC-labeled goat-anti-mouse IgG1 (Coulter-Immunotech).…”

Section: Facs Analysismentioning

confidence: 99%

T Cell Reactivity with Allergoids: Influence of the Type of APC

Kahlert

Grage-Griebenow

Stüwe

et al. 2000

The Journal of Immunology

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The use of allergoids for allergen-specific immunotherapy has been established for many years. The characteristic features of these chemically modified allergens are their strongly reduced IgE binding activity compared with the native form and the retained immunogenicity. T cell reactivity of chemically modified allergens is documented in animals, but in humans indirect evidence of reactivity has been concluded from the induction of allergen-specific IgG during immunotherapy. Direct evidence of T cell reactivity was obtained recently using isolated human T cells. To obtain further insight into the mechanism of action of allergoids, we compared the Ag-presenting capacity of different APC types, including DC and macrophages, generated from CD14+ precursor cells from the blood of grass pollen allergic subjects, autologous PBMC, and B cells. These APC were used in experiments together with Phl p 5-specific T cell clones under stimulation with grass pollen allergen extract, rPhl p 5b, and the respective allergoids. Using DC and macrophages, allergoids exhibited a pronounced and reproducible T cell-stimulating capacity. Responses were superior to those with PBMC, and isolated B cells failed to present allergoids. Considerable IL-12 production was observed only when using the DC for Ag presentation of both allergens and allergoids. The amount of IL-10 in supernatants was dependent on the phenotype of the respective T cell clone. High IL-10 production was associated with suppressed IL-12 production from the DC in most cases. In conclusion, the reactivity of Th cells with allergoids is dependent on the type of the APC.

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Characterization of Major Allergens of <i>Parietaria officinalis</i>

Cited by 15 publications

References 16 publications

Characterization of a Hypoallergenic Recombinant Bet v 1 Variant as a Candidate for Allergen-Specific Immunotherapy

Characterization of a Hypoallergenic Recombinant Bet v 1 Variant as a Candidate for Allergen-Specific Immunotherapy

Reactivity of T Cells with Grass Pollen Allergen Extract and Allergoid

T Cell Reactivity with Allergoids: Influence of the Type of APC

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