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Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
EPITOPE-BASED VACCINE FOR THE TREATMENT OF DER F 3 ALLERGY
Xiaodong Zhan1, Chaopin Li2, Yuxing Jiang3
1Deparment of Medical Parasitology, Wannan Medical College, Wuhu 241002, Anhui, People's Republic of China. College of life science, Anhui Normal University, Wuhu 241002, Anhui, People's Republic of China.. xdzhan@126.com.
Introduction:
mites allergic asthma is caused by exposure to home dust mite (HDM). Der f 3 is believed to be one of the major allergens in mites allergic asthma. The work was to identify the immune characteristics of Der f 3 epitope-based vaccine containing T cell and B cell epitopes.
Methods:
T cell lines were generated from peripheral blood mononuclear cells of Der f 3 allergic patients. Three T cell epitopes and five B cell epitopes of Der f 3, which we identified previously, were selected to design a polypeptide (named Der f3-peptides). DNA constructions encoding these Der f 3-peptides were expressed in Escherichia coli. The T cell lines were stimulated with the peptides and tested for proliferative capacity and cytokine production.
Results:
plasmid pET28a (+)-Der f 3-peptides was constructed and expressed in E. coli BL21, and the Der f3-peptides protein was purified and confirmed by Western blotting. The Der f 3-peptides were recognized by the T cell clones from allergic patients. SI value of Der f 3 group and Der f 3-peptides group were both higher than that of PBS group (P<0.05). The Der f 3 and Der f 3 peptides induced secretions of IL-4 and IL-5 were decreased compared with that of PBS group (P<0.05). The capacity of IgE-binding to Der f 3-peptides (41.25±5.67) μg/ml was decreased dramatically compared with that of Der f 3 (83.60 ± 10.92) μg/ml (P < 0.05).
Conclusions:
our results demonstrate that several major T cell epitopes and B cell epitopes of Der f 3 can be valuable for designing the peptide-based immunotherapeutics for the mites allergic asthma.
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