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Production of Egg Yolk Immunoglobulin (IgY) Against Recombinant LTB of Enterotoxigenic Escherichia coli (ETEC) and Evaluation of Its Protective Effect in Animal Model
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Maryam Mafi1    , Latif Mousavi gargari 2, Shahram Nazarian3 , Fatemeh Mohammad khani4 |
1- Microbial Biotechnology, Departmant of Cell Biology, Faculty of Basic Sciences, Shahed University, Tehran, Iran.
2- Ph.D Biochemistry, Departmant of Cell Biology, Faculty of Basic Sciences, Shahed University, Tehran, Iran , slmousavi@shahed.ac.ir
3- Ph.D. Microbiology, Biology Research Centere, Basic Science Faculty, Imam Hossein University, Tehran, Iran.
4- Ms.C Microbial Biotechnology, Departmant of Cell Biology, Faculty of Basic Sciences, Shahed University, Tehran, Iran. |
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Abstract: (1986 Views) |
ABSTRACT
Background and purpose:
Enterotoxigenic Escherichia coli is one of the most common bacterial causes of diarrhea worldwide.
One of the most important ETEC colonization factors is B subunit of the Heat Labile Enterotoxin, which forms the connecting part of the toxin. In the present study, egg yolk immunoglobulin (IgY) was produced and its effectiveness against recombinant LTB protein was tested on the animal model.
Material and methods:
Hens were injected intramuscularly with 100 μg of recombinant LTB protein. The antibody titer was estimated and the eggs were collected. The water soluble part of the yolk was separated and the antibody titer was subsequently estimated by ELISA.Effecacy of different concentrations of IgY on nutralising the effect of LT toxin on Y1 cells was also studied. In order to investigate the effect of IgY-treated bacteria on intestinal epithelial cells, the standard Ileal loop technique was used.
Results:
Protein expression led to the production of recombinant LTB showing molecular weight of 11 kDa on SDS PAGE. Immunization of hens induced serum antibody rise. The purified antibody was 144 mg per 12 ml of egg yolk. Purified IgY exhibited significant effect against recombinant LTB proteinAt the concentration of 125 μg per ml, the IgY could prevent the effects of Heat Labile Enterotoxin on Y1 cells. In the Ileal loop test, 1.5 mg / ml IgY neutralized the toxin effect of LTB on the intestine. The accumulation of fluid in the test loops decreased by 74.8% compared to the untreated control loops.
Conclusion:
The results showed that specific egg yolk immunoglobulin was effective against recombinant LTB protein and can be used as a preventive antibody to inhibit the Heat Labile Enterotoxin function of ETEC bacteria. |
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| Keywords: : Enterotoxigenic Escherichia coli, Heat labile enterotoxin, Passive immunity, Egg yolk immunoglobulin, Virulence factor |
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Full-Text [PDF 1419 kb]
(967 Downloads)
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Type of Study: Original Research |
Subject:
Biotechnology
Received: 2020/04/20 | Accepted: 2021/01/11 | Published: 2022/01/3
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