Reducing allergenicity by altering allergen fold: A mosaic protein of Phl p 1 for allergy vaccination

T. Ball, B. Linhart, K. Sonneck, K. Blatt, H. Herrmann, P. Valent, A. Stoecklinger, C. Lupinek, J. Thalhamer, A. A. Fedorov, Steven C. Almo, R. Valenta

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background: The major timothy grass pollen allergen, Phl p 1, resembles the allergenic epitopes of natural group I grass pollen allergens and is recognized by more than 95% of grass-pollen-allergic patients. Our objective was the construction, purification and immunologic characterization of a genetically modified derivative of the major timothy grass pollen allergen, Phl p 1 for immunotherapy of grass pollen allergy. Methods: A mosaic protein was generated by PCR-based re-assembly and expression of four cDNAs coding for Phl p 1 fragments and compared to the Phl p 1 wild-type by circular dichroism analysis, immunoglobulin E (IgE)-binding capacity, basophil activation assays and enzyme-linked immunosorbent assay competition assays. Immune responses to the derivative were studied in BALB/c mice. Results: Grass-pollen-allergic patients exhibited greater than an 85% reduction in IgE reactivity to the mosaic as compared with the Phl p 1 allergen and basophil activation experiments confirmed the reduced allergenic activity of the mosaic. It also induced less Phl p 1-specific IgE antibodies than Phl p 1 upon immunization of mice. However, immunization of mice and rabbits with the mosaic induced IgG antibodies that inhibited patients' IgE-binding to the wild-type allergen and Phl p 1-induced degranulation of basophils. Conclusion: We have developed a strategy based on rational molecular reassembly to convert one of the clinically most relevant allergens into a hypoallergenic derivative for allergy vaccination.

Original languageEnglish (US)
Pages (from-to)569-580
Number of pages12
JournalAllergy: European Journal of Allergy and Clinical Immunology
Volume64
Issue number4
DOIs
StatePublished - Apr 2009

Fingerprint

Allergens
Pollen
Hypersensitivity
Vaccination
Poaceae
Immunoglobulin E
Basophils
Phleum
Proteins
Immunization
Seasonal Allergic Rhinitis
Antibodies
Circular Dichroism
Immunotherapy
Epitopes
Complementary DNA
Immunoglobulin G
Enzyme-Linked Immunosorbent Assay
Rabbits
Polymerase Chain Reaction

Keywords

  • Allergens
  • Immunotherapy
  • Pollens

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Reducing allergenicity by altering allergen fold : A mosaic protein of Phl p 1 for allergy vaccination. / Ball, T.; Linhart, B.; Sonneck, K.; Blatt, K.; Herrmann, H.; Valent, P.; Stoecklinger, A.; Lupinek, C.; Thalhamer, J.; Fedorov, A. A.; Almo, Steven C.; Valenta, R.

In: Allergy: European Journal of Allergy and Clinical Immunology, Vol. 64, No. 4, 04.2009, p. 569-580.

Research output: Contribution to journalArticle

Ball, T, Linhart, B, Sonneck, K, Blatt, K, Herrmann, H, Valent, P, Stoecklinger, A, Lupinek, C, Thalhamer, J, Fedorov, AA, Almo, SC & Valenta, R 2009, 'Reducing allergenicity by altering allergen fold: A mosaic protein of Phl p 1 for allergy vaccination', Allergy: European Journal of Allergy and Clinical Immunology, vol. 64, no. 4, pp. 569-580. https://doi.org/10.1111/j.1398-9995.2008.01910.x
Ball, T. ; Linhart, B. ; Sonneck, K. ; Blatt, K. ; Herrmann, H. ; Valent, P. ; Stoecklinger, A. ; Lupinek, C. ; Thalhamer, J. ; Fedorov, A. A. ; Almo, Steven C. ; Valenta, R. / Reducing allergenicity by altering allergen fold : A mosaic protein of Phl p 1 for allergy vaccination. In: Allergy: European Journal of Allergy and Clinical Immunology. 2009 ; Vol. 64, No. 4. pp. 569-580.
@article{b48dfbe2d4164430a4e10543ae639fdc,
title = "Reducing allergenicity by altering allergen fold: A mosaic protein of Phl p 1 for allergy vaccination",
abstract = "Background: The major timothy grass pollen allergen, Phl p 1, resembles the allergenic epitopes of natural group I grass pollen allergens and is recognized by more than 95{\%} of grass-pollen-allergic patients. Our objective was the construction, purification and immunologic characterization of a genetically modified derivative of the major timothy grass pollen allergen, Phl p 1 for immunotherapy of grass pollen allergy. Methods: A mosaic protein was generated by PCR-based re-assembly and expression of four cDNAs coding for Phl p 1 fragments and compared to the Phl p 1 wild-type by circular dichroism analysis, immunoglobulin E (IgE)-binding capacity, basophil activation assays and enzyme-linked immunosorbent assay competition assays. Immune responses to the derivative were studied in BALB/c mice. Results: Grass-pollen-allergic patients exhibited greater than an 85{\%} reduction in IgE reactivity to the mosaic as compared with the Phl p 1 allergen and basophil activation experiments confirmed the reduced allergenic activity of the mosaic. It also induced less Phl p 1-specific IgE antibodies than Phl p 1 upon immunization of mice. However, immunization of mice and rabbits with the mosaic induced IgG antibodies that inhibited patients' IgE-binding to the wild-type allergen and Phl p 1-induced degranulation of basophils. Conclusion: We have developed a strategy based on rational molecular reassembly to convert one of the clinically most relevant allergens into a hypoallergenic derivative for allergy vaccination.",
keywords = "Allergens, Immunotherapy, Pollens",
author = "T. Ball and B. Linhart and K. Sonneck and K. Blatt and H. Herrmann and P. Valent and A. Stoecklinger and C. Lupinek and J. Thalhamer and Fedorov, {A. A.} and Almo, {Steven C.} and R. Valenta",
year = "2009",
month = "4",
doi = "10.1111/j.1398-9995.2008.01910.x",
language = "English (US)",
volume = "64",
pages = "569--580",
journal = "Allergy: European Journal of Allergy and Clinical Immunology",
issn = "0105-4538",
publisher = "Wiley-Blackwell",
number = "4",

}

TY - JOUR

T1 - Reducing allergenicity by altering allergen fold

T2 - A mosaic protein of Phl p 1 for allergy vaccination

AU - Ball, T.

AU - Linhart, B.

AU - Sonneck, K.

AU - Blatt, K.

AU - Herrmann, H.

AU - Valent, P.

AU - Stoecklinger, A.

AU - Lupinek, C.

AU - Thalhamer, J.

AU - Fedorov, A. A.

AU - Almo, Steven C.

AU - Valenta, R.

PY - 2009/4

Y1 - 2009/4

N2 - Background: The major timothy grass pollen allergen, Phl p 1, resembles the allergenic epitopes of natural group I grass pollen allergens and is recognized by more than 95% of grass-pollen-allergic patients. Our objective was the construction, purification and immunologic characterization of a genetically modified derivative of the major timothy grass pollen allergen, Phl p 1 for immunotherapy of grass pollen allergy. Methods: A mosaic protein was generated by PCR-based re-assembly and expression of four cDNAs coding for Phl p 1 fragments and compared to the Phl p 1 wild-type by circular dichroism analysis, immunoglobulin E (IgE)-binding capacity, basophil activation assays and enzyme-linked immunosorbent assay competition assays. Immune responses to the derivative were studied in BALB/c mice. Results: Grass-pollen-allergic patients exhibited greater than an 85% reduction in IgE reactivity to the mosaic as compared with the Phl p 1 allergen and basophil activation experiments confirmed the reduced allergenic activity of the mosaic. It also induced less Phl p 1-specific IgE antibodies than Phl p 1 upon immunization of mice. However, immunization of mice and rabbits with the mosaic induced IgG antibodies that inhibited patients' IgE-binding to the wild-type allergen and Phl p 1-induced degranulation of basophils. Conclusion: We have developed a strategy based on rational molecular reassembly to convert one of the clinically most relevant allergens into a hypoallergenic derivative for allergy vaccination.

AB - Background: The major timothy grass pollen allergen, Phl p 1, resembles the allergenic epitopes of natural group I grass pollen allergens and is recognized by more than 95% of grass-pollen-allergic patients. Our objective was the construction, purification and immunologic characterization of a genetically modified derivative of the major timothy grass pollen allergen, Phl p 1 for immunotherapy of grass pollen allergy. Methods: A mosaic protein was generated by PCR-based re-assembly and expression of four cDNAs coding for Phl p 1 fragments and compared to the Phl p 1 wild-type by circular dichroism analysis, immunoglobulin E (IgE)-binding capacity, basophil activation assays and enzyme-linked immunosorbent assay competition assays. Immune responses to the derivative were studied in BALB/c mice. Results: Grass-pollen-allergic patients exhibited greater than an 85% reduction in IgE reactivity to the mosaic as compared with the Phl p 1 allergen and basophil activation experiments confirmed the reduced allergenic activity of the mosaic. It also induced less Phl p 1-specific IgE antibodies than Phl p 1 upon immunization of mice. However, immunization of mice and rabbits with the mosaic induced IgG antibodies that inhibited patients' IgE-binding to the wild-type allergen and Phl p 1-induced degranulation of basophils. Conclusion: We have developed a strategy based on rational molecular reassembly to convert one of the clinically most relevant allergens into a hypoallergenic derivative for allergy vaccination.

KW - Allergens

KW - Immunotherapy

KW - Pollens

UR - http://www.scopus.com/inward/record.url?scp=62449174319&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=62449174319&partnerID=8YFLogxK

U2 - 10.1111/j.1398-9995.2008.01910.x

DO - 10.1111/j.1398-9995.2008.01910.x

M3 - Article

C2 - 19243361

AN - SCOPUS:62449174319

VL - 64

SP - 569

EP - 580

JO - Allergy: European Journal of Allergy and Clinical Immunology

JF - Allergy: European Journal of Allergy and Clinical Immunology

SN - 0105-4538

IS - 4

ER -