EPITOPE-BASED VACCINE FOR THE TREATMENT OF DER F 3 ALLERGY.

VACUNA BASADA EN EPÍTOPO PARA EL TRATAMIENTO DE LA ALERGIA DER F 3.

ALLERGIES; EPITOPES; ALLERGENS; VACCINES; DERMATOPHAGOIDES; T cells; IMMUNOTHERAPY; VACCINATION; RESPIRATORY allergy; AMINO acids; ANIMAL experimentation; ANTIGENS; CELL lines; CELL physiology; COMPARATIVE studies; CYTOKINES; RESEARCH methodology; MEDICAL cooperation; MITES; NUCLEOTIDES; RESEARCH; EVALUATION research; PREVENTION

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. [ABSTRACT FROM AUTHOR]

Introducción: el asma alergica esta causada por la exposicion a los acaros del polvo casero (HDM). Der f 3 se cree que es uno de los principales alergenos en los acaros del asma alergica. El trabajo consistio en identificar las caracteristicas inmunologicas de la vacuna basada en epitopo-Der f 3 que contienen las celulas T y las celulas B. Métodos: se generaron lineas de celulas T a partir de celulas mononucleares de sangre periferica de pacientes alergicos a Der f 3. Tres epitopos de celulas T y cinco epitopes de celulas B de Der f 3, que hemos identificado previamente, fueron seleccionados para disenar un polipeptido (denominados peptidos Der f 3). Construcciones de DNA que codifican estos peptidos Der f 3 se expresaron en Escherichia coli. Las lineas de celulas T se estimularon con los peptidos y se utilizaron en el ensayo por su capacidad proliferativa y la produccion de citoquinas. Resultados: el plasmido pET28a (+) - Der f 3-peptidos se construyo y se expresaron en E. coli BL21, y la proteina de Der f 3-peptidos se purifico y se confirmaron mediante transferencia de Western. Los Der f 3-peptidos fueron reconocidos por los clones de celulas T procedentes de pacientes alergicos. Valor SI de Der f 3 grupo y f grupo 3-peptidos Der eran tanto mayor que la del grupo de PBS (P <0, 05). El Der f 3 y Der f 3 peptidos indujeron secreciones de IL-4 e IL-5 se redujeron en comparacion con la del grupo PBS (P <0,05). La capacidad de union a IgE a Der f 3-peptidos (41, 25 ± 5, 67) μg/ml se redujo drasticamente en comparacion con el de Der f 3 (83,60 ± 10,92) μg/ml (P <0,05). Conclusiones: nuestros resultados demuestran que varios de los principales epitopos de celulas T y de celulas B de Der f 3 pueden ser valiosos para el diseno de agentes inmunoterapeuticos basados en peptidos para los acaros del asma alergica. [ABSTRACT FROM AUTHOR]

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