Bet v 1-displaying elastin-like polypeptide nanoparticles induce a strong humoral and weak CD4+ T-cell response against Bet v 1 in a murine immunogenicity model

Frontiers in Immunology, vol. 13, pp. 1006776

Abstract

There is growing concern about the toxicity of colloidal aluminum salts used as adjuvants in subcutaneous allergen immunotherapy (SCIT). Therefore, alternative adjuvants and delivery systems are being explored to replace alum in SCIT. We applied micellar elastin-like polypeptides (ELPs), a type of self-assembling protein, to replace alum as vaccine adjuvant in birch pollen SCIT. ELP and an ELP-Bet v 1 fusion protein were expressed inE. coliand purified by immuno-affinity chromatography and inverse-transition cycling (ITC). Nanoparticles self-assembled from ELP and a 9:1 ELP/ELP-Bet v 1 mixture were characterized by using dynamic light scattering and atomic force microscopy. Allergenicity was assessed by measuring mediator release from rat basophilic leukemia cells transformed with the human FcϵR1 and sensitized with sera derived from human birch pollen allergic patients. Humoral and T-cell immunity were investigated by immunizing naïve mice with the ELP/ELP-Bet v 1 nanoparticles or alum-adsorbed Bet v 1, both containing 36 µg Bet v 1. ELP and ELP/ELP-Bet v 1 self-assembled at 37°C into spherically shaped micelles with a diameter of ~45 nm. ELP conjugation made Bet v 1 hypo-allergenic (10-fold). Compared to alum-adsorbed Bet v 1, ELP/ELP-Bet v 1 nanoparticles induced stronger IgG responses with an earlier onset. Additionally, ELP/ELP-Bet v 1 did not induce Th2 skewing cytokines and IgE. The hypoallergenic character and strong humoral immune response in the absence of a Th2-skewing T-cell response make ELP-based nanoparticles a promising candidate to replace alum in SCIT.

Authors 15

1. Jolinde van Strien Aachen Department of Supramolecular and Biomaterials Chemistry

   Leiden University

   Affiliation as printed

   Department of Supramolecular and Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands

2. Hans Warmenhoven Aachen R&D Department

   HAL Allergy (Netherlands) · Amsterdam University Medical Centers

   Affiliation as printed

   Department of Experimental Immunology, Amsterdam University Medical Centers, Amsterdam, Netherlands

   R&D Department, Haarlems Allergenen Laboratorium (HAL) Allergy B.V., Leiden, Netherlands

3. Adrian Logiantara

   Amsterdam University Medical Centers

   Affiliation as printed

   Department of Experimental Immunology, Amsterdam University Medical Centers, Amsterdam, Netherlands

4. Max Makurat Aachen Department of Supramolecular and Biomaterials Chemistry

   Leiden University

   Affiliation as printed

   Department of Supramolecular and Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands

5. Lorenz Aglas

   University of Salzburg

   Affiliation as printed

   Division of Allergy and Immunology, Department of Biosciences, Paris Lodron University of Salzburg, Salzburg, Austria

6. Athanasios Bethanis

   University of Salzburg

   Affiliation as printed

   Division of Allergy and Immunology, Department of Biosciences, Paris Lodron University of Salzburg, Salzburg, Austria

7. Romain Leboux Aachen Department of BioTherapeutics Leiden Academic Centre for Drug Research

   Leiden University

   Affiliation as printed

   Department of BioTherapeutics, Leiden Academic Centre for Drug Research (LACDR), Leiden University, Leiden, Netherlands

8. Leonie S. van Rijt

   Amsterdam University Medical Centers

   Affiliation as printed

   Department of Experimental Immunology, Amsterdam University Medical Centers, Amsterdam, Netherlands

9. John Andrew MacKay

   University of Southern California

   Affiliation as printed

   Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, CA, United States

10. Johannes W. van Schijndel Aachen R&D Department

    HAL Allergy (Netherlands)

    Affiliation as printed

    R&D Department, Haarlems Allergenen Laboratorium (HAL) Allergy B.V., Leiden, Netherlands

11. Grégory F. Schneider Aachen Department of Supramolecular and Biomaterials Chemistry

    Leiden University

    Affiliation as printed

    Department of Supramolecular and Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands

12. René C. L. Olsthoorn Aachen Department of Supramolecular and Biomaterials Chemistry

    Leiden University

    Affiliation as printed

    Department of Supramolecular and Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands

13. Wim Jiskoot Aachen Department of BioTherapeutics Leiden Academic Centre for Drug Research

    Leiden University

    Affiliation as printed

    Department of BioTherapeutics, Leiden Academic Centre for Drug Research (LACDR), Leiden University, Leiden, Netherlands

14. Ronald van Ree

    Amsterdam University Medical Centers

    Affiliation as printed

    Department of Experimental Immunology, Amsterdam University Medical Centers, Amsterdam, Netherlands

    Department of Otorhinolaryngology, Amsterdam University Medical Centers, Amsterdam, Netherlands

15. Alexander Kros corresponding Aachen Department of Supramolecular and Biomaterials Chemistry

    Leiden University

    Affiliation as printed

    Department of Supramolecular and Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands