Detection of allergen composition and in vivo immunogenicity of depigmented allergoids of Betula alba

Carnés, Jerónimo; Himly, Martin; Gallego, Mayte; Iraola, Víctor; Robinson, Douglas S.; Fernández-Caldas, E; Briza, Peter

doi:10.1111/j.1365-2222.2008.03132.x

Cited by 37 publications

(34 citation statements)

References 22 publications

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“…However, this property also reduces binding of antibodies used in assays to detect allergen proteins and has hampered assessment of the allergen content of allergoids. Recently, we described the application of mass spectroscopy-based peptide mapping to determine allergen sequences in Dpg-Pol extracts of Betula alba [13]. Using these techniques in the present study, allergen sequences of the major allergens Der p 1 and Der p 2 and minor allergens (Der p 3, 4, 8, 10, 11 and 20) were detected in proteolytic digests of Dpg-Pol extracts of D. pteronyssinus.…”

Section: Discussionmentioning

confidence: 83%

“…One difficulty with assessing allergoids is that the polymerisation process blocks access to antibodies, making it difficult to use antibody-based assays to measure allergen content in these vaccines. Recently, we described the use of mass spectroscopic techniques to confirm preservation of major allergen content in Dpg-Pol birch pollen extracts [13], which may allow the development of methods for standardisation of allergoids [14]. …”

Section: Introductionmentioning

confidence: 99%

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Depigmented and Polymerised House Dust Mite Allergoid: Allergen Content, Induction of IgG4 and Clinical Response

Gallego

Iraola

Himly

et al. 2010

Int Arch Allergy Immunol

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Background: Polymerised allergenic extracts (allergoids) are commonly used in allergen immunotherapy. Clinical efficacy and safety of these extracts have been demonstrated. Recently, allergen sequences have been identified by mass spectrometry in depigmented and polymerised (Dpg-Pol) extracts. The objectives of this study were to investigate the presence of allergens in Dpg-Pol extracts of house dust mite and to analyze the immunological changes induced by these extracts in asthmatic patients enrolled in a double-blind, placebo-controlled study. Methods: Dpg-Pol extracts were manufactured and vaccines with a composition of 50% Dermatophagoides pteronyssinus and 50% D. farinae (100 HEPL/ml) were prepared. Allergen composition was analyzed by mass spectrometry. Patients with asthma and rhinoconjunctivitis were treated in a 1-year, double-blind, placebo-controlled, parallel-group study with 6 up-dosing and monthly maintenance injections. Specific IgE and IgG4 titres to D. pteronyssinus, Der p 1 and Der p 2 were measured in patients’ sera using the CAP system and direct ELISA experiments. Results: Sequences from the major allergens Der p 1 and Der p 2 and from other allergens were identified in native and Dpg-Pol extracts. There was a statistically significant increase in specific IgG4, a decrease in the ratio of IgE/IgG4 to D. pteronyssinus and a significant increase in specific IgG4 to Der p 1 and Der p 2 in the patients allotted to active treatment. Conclusions: The detection of allergen sequences suggests preservation of major and minor allergens in Dpg-Pol allergoids from house dust mites. Efficacy in asthma treatment and the increase in specific IgG4 seem to be associated with the presence of major allergens in Dpg-Pol allergen extracts.

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Section: Discussionmentioning

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Depigmented and Polymerised House Dust Mite Allergoid: Allergen Content, Induction of IgG4 and Clinical Response

Gallego

Iraola

Himly

et al. 2010

Int Arch Allergy Immunol

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“…One of the techniques used for extract modification is polymerisation with glutaraldehyde, a process that modifies IgE epitopes, resulting in a safer vaccine that allows the administration of increased concentrations of allergen per dose. These formulations are known as allergoids and are composed of high molecular weight molecules that include all the individual allergens present in native (N) extracts forming proteic macrocomplexes [4]. A further step in this sense implies the production of allergoids using highly purified extracts.…”

Section: Introductionmentioning

confidence: 99%

Modulation of the Humoral Response to Dermatophagoides pteronyssinus Allergens in BALB/c Mice by Extract Modification and Adjuvant Use

Coaña

Carnés²,

Gallego³

et al. 2011

Int Arch Allergy Immunol

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Background: Currently, several strategies are being used in order to improve the safety and efficacy of allergen-specific immunotherapy; these strategies include the use of modified hypoallergenic extracts as well as different adjuvants with immunomodulatory properties in combination with native or modified extracts. The objectives of this study were to investigate the humoral response generated in mice to modified Dermatophagoides pteronyssinus extracts in the presence or absence of two different adjuvants. Methods: BALB/c mice were inoculated either with native, depigmented or depigmented-polymerised D. pteronyssinus without adjuvants or combined with aluminium hydroxide or oligodeoxinucleotides containing CpG motifs. IgE concentration, specific total IgG, IgG1 and IgG2a titres were measured in mice sera and cross-reactivity inhibition experiments were performed. IgG antigenic profiles were obtained by immunoblotting for all formulations. Results: Inoculation of depigmented-polymerised extract induced statistically significant lower IgE levels than the native extract even when adsorbed onto aluminium hydroxide. When this extract was inoculated in the presence of oligodeoxinucleotides containing CpG motifs, it elicited high IgG levels, a high IgG2a/lgG1 ratio and low IgE production. Furthermore, the antigenic profiles observed after extract inoculation showed punctual differences between the depigmented-polymerised extract and the native or depigmented extracts. Conclusions: Our results suggest that the depigmentation and polymerisation process modifies the native extract’s antigenic and immunogenic properties and converts the depigmented-polymerised extract into a better choice for allergen-specific immunotherapy.

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“…Also, some allergies are caused by sensitization to a few proteins or even a single molecule, whereas others are caused by diverse and complex allergen expression [46,47].…”

Section: Discussionmentioning

confidence: 99%

Comparison of Skin Prick Tests with In Vitro Allergy Tests in the Characterization of Horses with Recurrent Airway Obstruction

Tilley¹,

Luis²,

Ferreira³

2012

Journal of Equine Veterinary Science

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Detection of allergen composition and in vivo immunogenicity of depigmented allergoids of Betula alba

Cited by 37 publications

References 22 publications

Depigmented and Polymerised House Dust Mite Allergoid: Allergen Content, Induction of IgG4 and Clinical Response

Depigmented and Polymerised House Dust Mite Allergoid: Allergen Content, Induction of IgG4 and Clinical Response

Modulation of the Humoral Response to <i>Dermatophagoides pteronyssinus </i>Allergens in BALB/c Mice by Extract Modification and Adjuvant Use

Comparison of Skin Prick Tests with In Vitro Allergy Tests in the Characterization of Horses with Recurrent Airway Obstruction

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