A hypoallergenic variant of Der p 1 as a candidate for mite allergy vaccines

Walgraffe, David; Mattéotti, Christel; Bakkoury, Mohamed El; Garcia, Lida; Marchand, Céline; Bullens, Dominique; Vandenbranden, Michel; Jacquet, Alain

doi:10.1016/j.jaci.2008.11.038

Cited by 31 publications

(28 citation statements)

References 38 publications

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“…Noteworthy, if Der p 2 alone has been previously successfully refolded from inclusion bodies [13], this has not yet been achieved for Der p 1. Whereas the proregion of cysteine proteases is critical for the folding of such enzymes [33,34], recent data suggest that recombinant proDer p 1 remains prone to aggregation [35], indicating that the presence of the propeptide per se is not sufficient to ensure natural conformation. Although not shown, our preliminary refolding data on F2 and F4 suggest that such fusion proteins tend to remain insoluble and form aggregates.…”

Section: Discussionmentioning

confidence: 99%

Recombinant Fusion Proteins Assembling Der p 1 and Der p 2 Allergens from <i>Dermatophagoides pteronyssinus</i>

Bussières

Floch²,

Bulder³

et al. 2010

Int Arch Allergy Immunol

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Background: Fusion proteins assembling multiple allergens can be engineered by recombinant DNA technologies in order to produce tools for diagnostic and immunotherapeutic purposes. Herein, we developed and characterized chimeras assembling Der p 1 and Der p 2 allergens as potential candidate vaccines against house dust mite allergy. Methods: Fusion proteins encompassing Der p 2 with either mature or proDer p 1 were expressed in Escherichia coli or Pichia pastoris. Forms with mutation in Der p 1 catalytic site were also engineered. Purified chimeras were characterized by immunoblotting, circular dichroism, disulfide bond mapping, basophil and T lymphocyte stimulation assays. Results: Four fusion proteins were expressed in E. coli as inclusion bodies, whereas only chimeras comprising proDer p 1 were obtained in yeast. All such hybrids formed polymers and aggregates, and yeast-expressed chimeras were unstable. Circular dichroism analysis performed after refolding of bacteria expressed chimeras encompassing mature Der p 1 confirmed partial folding, consistent with the occurrence of both correct and inappropriate intramolecular disulfide bonds. All fusion molecules were recognized by Der p 1- and Der p 2-specific human IgEs, monoclonal and polyclonal antibodies. Fusion proteins activate basophils from mite-allergic patients and trigger the proliferation of specific CD4+ T cells, albeit to a lower level when compared to individual allergens. Conclusions: Production of multiple Der p 1-Der p 2 fusion proteins exhibiting partial folding and proper antigenic properties has been achieved. Nonetheless, significant solubility and stability issues currently limit the application of such chimeras for immunotherapy or diagnostic.

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Section: Discussionmentioning

confidence: 99%

Recombinant Fusion Proteins Assembling Der p 1 and Der p 2 Allergens from <i>Dermatophagoides pteronyssinus</i>

Bussières

Floch²,

Bulder³

et al. 2010

Int Arch Allergy Immunol

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“…Our study on IgE binding and competition, however, indicates that targeting the 4 residues with less conservative substitutions could be a strategy for developing hypoallergic Der p 2. The T-cell repertoire to Der p 2 is complex with individuals responding to several epitopes, 23 so variants made hypoallergenic by a few amino acid substitution would be expected to retain most of their T-cell reactivity as in folding variants, such as those described for Der p 1 24 that could lose some epitopes by altering antigen processing. This would need to be tested as per the Der p 1 study.…”

Section: Discussionmentioning

confidence: 99%

“…This would need to be tested as per the Der p 1 study. 24 Studies on IgE binding and competition also show that it would be important to determine which of the Der p 2 variants would induce IgG antibodies that can block IgE binding to the variants in the environment. This would be critical for immunotherapy based on the strategy for inducing IgG-blocking antibodies.…”

Section: Discussionmentioning

confidence: 99%

Effect of Amino Acid Polymorphisms of House Dust Mite Der p 2 Variants on Allergic Sensitization

Tanyaratsrisakul

Jirapongsananuruk

Kulwanich

et al. 2016

Allergy Asthma Immunol Res

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“…An ideal vaccine for desensitization ought to be immunogenic, while being nonallergenic [62,63]. In order to evaluate the mutants’ immunogenicity, the proliferation of T lymphocytes from allergic individuals induced by the two mutants was compared with the proliferation induced by Mus m 1.…”

Section: Discussionmentioning

confidence: 99%

In Search of a Vaccine for Mouse Allergy: Significant Reduction of Mus m 1 Allergenicity by Structure-Guided Single-Point Mutations

Ferrari

Breda

Longhi

et al. 2011

Int Arch Allergy Immunol

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Background: Mouse urinary proteins are relevant allergens from mice urine. We used the recombinant protein Mus m 1 as an allergen model to identify if, by altering Mus m 1 architecture via single-point mutations, we could effectively modify its allergenicity. Methods: Based on structural considerations, we synthesized two single-point mutants, Mus m 1-Y120A and Mus m 1-Y120F, which were expected to harbor large structural alterations. Circular dichroism and fluorescence analysis showed significant conformational rearrangements of the aromatic side chains in the internal cavity of Mus m 1-Y120A when compared to Mus m 1-Y120F and Mus m 1. Evaluation of the allergenic potential of the recombinant molecules was performed in vitro with both immunochemical approaches and assays based on the measurement of basophil degranulation. Moreover, to assess the integrity of the T cell epitopes and as an in vitro measure of immunogenicity, we tested the reactivity of T lymphocytes from subjects allergic to mouse urine against proteins and synthetic peptides encompassing the immunodominant linear epitope containing the mutation. Results: We found that the selected point mutation was able to modulate the protein allergenicity, and to severely impair the recognition of Mus m 1 by IgE, while T cell reactivity was fully maintained. Conclusions: In silico predicted, minimum selected structural modifications allowed to design one protein with reduced allergenicity and preserved immunogenicity. Structurally guided mutations can direct the design of proteins with reduced allergenicity which can be used as vaccines for a safer and more effective immunotherapy of allergic disorders.

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A hypoallergenic variant of Der p 1 as a candidate for mite allergy vaccines

Cited by 31 publications

References 38 publications

Recombinant Fusion Proteins Assembling Der p 1 and Der p 2 Allergens from <i>Dermatophagoides pteronyssinus</i>

Recombinant Fusion Proteins Assembling Der p 1 and Der p 2 Allergens from <i>Dermatophagoides pteronyssinus</i>

Effect of Amino Acid Polymorphisms of House Dust Mite Der p 2 Variants on Allergic Sensitization

In Search of a Vaccine for Mouse Allergy: Significant Reduction of Mus m 1 Allergenicity by Structure-Guided Single-Point Mutations

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