Functional Deimmunization of Interferon Beta-1b by Identifying and Silencing Human T Cells Epitopes.

Functional Deimmunization of Interferon Beta-1b by Identifying and Silencing Human T Cells Epitopes. Iran J Allergy Asthma Immunol. 2019 Aug 17;18(4):427-440 Authors: Moradi Hasan-Abad A, Adabi E, Sadroddiny E, Khorramizadeh MR, Mazlomi MA, Mehravar S, Kardar GA Abstract Interferonbeta-1b (IFNβ-1b) developed as therapeutic protein for the treatment of multiple sclerosis (MS). Studies have been shown that Long-term usage of this protein can lead to the development of anti-drug antibodies (ADAs) and this phenomenon cause total loss or reduced efficacy of IFNβ-1b. The aim of this study was to predict and silence IFNβ-1b T-cells epitopes by in silico methods and genetic engineering. Based on bioinformatics studies we identified optimal sets of conservative point mutations for eliminating T-cells epitopes in IFNβ-1b protein. Four synthetic genes with desirable mutation constructed and PET26b+ was used as an expression vector in E. coli. The expression of this proteins confirmed by SDS-PAGE and Western blotting, consequently, IFNβ-1b proteins was purified by His-tag chromatography. To determined activity of mutants' variants anti-proliferative and anti-viral activity compared to wild form was evaluated using MTT assay in A549 and Vero cells lines respectively. Also the immunogenicity of mutant proteins compared with Betaseron measured in BALB/c mice. The in vitro bioactivity analysis demonstrated that functional activities of all muta...
Source: Iranian Journal of Allergy, Asthma and Immunology - Category: Allergy & Immunology Authors: Tags: Iran J Allergy Asthma Immunol Source Type: research