Expression, purification and epitope analysis of Pla a 3 allergen from Platanus acerifolia pollen

Mol Med Rep. 2017 Sep;16(3):2851-2855. doi: 10.3892/mmr.2017.6867. Epub 2017 Jun 28.

Abstract

Platanus acerifolia (P. acerifolia) is an important cause of pollinosis in cities. The use of allergen extracts on patients with allergic diseases is the most commonly applied method to attempt to treat pollinosis. Pla a 3, a non‑specific lipid transfer protein, is a major allergen present in P. acerifolia pollen extracts. In the present study, the Pla a 3 gene was sub‑cloned into a pSUMO‑Mut vector using Stu I and Xho I sites and transformed into the Arctic Express™ (DE3) RP E. coli host strain. The purified Pla a 3 allergen was analyzed by western blotting and the results revealed that the Pla a 3 allergen has the ability to bind IgE in the P. acerifolia pollen of allergic patients' sera. Moreover, the authors predicted the potential B cell epitopes of the Pla a 3 allergen using the DNAStar Protean system, the Bioinformatics Predicted Antigenic Peptides system and the BepiPred 1.0 server. In addition, the T cell epitopes were predicted by the SYFPEITHI database and the NetMHCII‑2.2 server. As a result, two B cell epitopes (35‑45 and 81‑86) and four potential T cell epitopes including 2‑15, 45‑50, 55‑61 and 67‑73 were predicted in the present study. The current results can be used to contribute to allergen immunotherapies and useful in peptide‑based vaccine designs of pollen allergy.

MeSH terms

  • Adolescent
  • Adult
  • Antigens, Plant / chemistry
  • Antigens, Plant / genetics
  • Antigens, Plant / immunology
  • Antigens, Plant / isolation & purification
  • Cloning, Molecular
  • Epitopes, B-Lymphocyte / chemistry
  • Epitopes, B-Lymphocyte / genetics
  • Epitopes, B-Lymphocyte / immunology*
  • Epitopes, B-Lymphocyte / isolation & purification
  • Epitopes, T-Lymphocyte / chemistry
  • Epitopes, T-Lymphocyte / genetics
  • Epitopes, T-Lymphocyte / immunology*
  • Epitopes, T-Lymphocyte / isolation & purification
  • Escherichia coli / genetics
  • Female
  • Humans
  • Immunoglobulin E / blood
  • Immunoglobulin E / immunology*
  • Magnoliopsida / chemistry
  • Magnoliopsida / genetics
  • Magnoliopsida / immunology*
  • Male
  • Middle Aged
  • Models, Molecular
  • Pollen / chemistry
  • Pollen / genetics
  • Pollen / immunology
  • Protein Conformation
  • Rhinitis, Allergic, Seasonal / blood
  • Rhinitis, Allergic, Seasonal / immunology*
  • Young Adult

Substances

  • Antigens, Plant
  • Epitopes, B-Lymphocyte
  • Epitopes, T-Lymphocyte
  • Immunoglobulin E