Neutralizing Potential of Fab IgG Hybrid Antibody Against Dengue Virus (DENV-1,2,3,4) Expressed on Mesenchymal Stem Cells

Year 2018, Volume: 44 Issue: 1, 12 - 19, 16.01.2018

Fedik A. Rantam Rahaju Ernawatı Jola Rahmahanı Eryk Hendrıanto Helen Susılowatı Rofiqul A’la Rizki Kriestya Mayasarı

Abstract

Dengue is an acute mosquito born viral
infections in tropical and subtropical countries, that has been steadily
increasing every year. Dengue haemorrhagic fever in Indonesia is caused by all
4 viral serotypes termed as DENV-1, DENV-2, DENV-3, and DENV-4. It is very
difficult to treat the disease and there is no effective vaccine to eradicate
Dengue virus infections .The aim of this research is to express the antibody
fragment gene coding VL and VH of Fab IgG antibody in rat bone marrow
mesenchymal stem cells (rat BM-MSCs). The genes were isolated from immunized
mice using inactive virus of all dengue serotypes. Then genes were hybridized
through ligation and insertion into plasmid pBR322, and transfected into rat BMMSCs.
To analytically characterize Fab IgG hybrid binding ability, capacity and
specificity, we used immune precipitation, western blotting, neutralization
assay, and ELISA. All assays suggested that hybrid Fab IgG antibody have high
reactivity and affinity, to efficiently neutralize all Dengue virus serotypes
1,2,3,4. Results of this study show that hybrid Fab IgG antibody can be used as
neutralizing agent for the treatment of dengue infections in the future. 

Keywords

Dengue virus, FabIgG hybrid antibody, generated expression ratBMMSCs, reactivity, neutralization

Mezenkimal Kök Hücreler Üzerinde Dengue Virüsüne Karşı (DENV-1,2,3,4) Fab IgG Hibrit Antikorunun Nötralize Edici Potansiyeli

Abstract

Dengue humması, tropik ve subtropikal
ülkelerde sivrisineklerden kaynaklanan akut viral bir enfeksiyon olup, her yıl
giderek artmaktadır. Endonezya’daki Dang
kanamalı ateşine DENV-1, DENV-2, DENV-3 ve DENV-4 adı verilen 4 viral serotip
neden olmaktadır. Hastalığın tedavisi çok zordur ve Dang virüsü
enfeksiyonlarını yok etmek için etkili bir aşı yoktur. Bu araştırmanın amacı,
sıçan kemik iliği mezenkimal kök hücrelerinde (sıçan BM-MSCs) bulunan Fab IgG
antikorunun VL ve VH antikor fragmentini kodlayan geninin açığa çıkarılmasıdır.
Genler, tüm dang serotiplerinin inaktif virüsü kullanılarak aşılanmış
farelerden izole edilmiştir. Sonrasında genler, ligasyon ve plasmid pBR322
içine gömülmesi yoluyla hibridize edilmiş ve sıçan BMMSC’lerine nakledilmiştir. Analitik olarak Fab IgG’nin hibrid bağlama yeteneği, kapasitesi ve özgünlüğünü karakterize
etmek için; bağışıklık çökeltmesi, western blotting, nötralizasyon testi ve
ELISA kullanılmıştır. Bütün deneyler, hibrid Fab IgG antikorunun tüm Dang
virüsü serotipleri 1,2,3,4’ü etkili
bir şekilde nötrleştirmek için yüksek reaktiviteye ve afiniteye sahip olduğunu
ortaya koymaktadır. Bu çalışmanın sonuçları, melez Fab IgG antikorunun
gelecekte dang enfeksiyonlarının tedavisinde nötralize edici ajan olarak
kullanılabileceğini göstermektedir.

Keywords

Dengue virüsü, Fab IgG hibrid antikor, sıçan kemik iliği mezenkimal kök hücrelerinin ekspresyonu (rat- BMMSC), reaktivite, nötrleştirme

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