Evaluation of the prophylactic effect of egg yolk antibody (IgY) produced against the recombinant protein containing IpaD, IpaB, StxB, and VirG proteins from Shigella.

Autor: Felegary A; Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran., Nazarian S; Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran. Electronic address: nazarian@ihu.ac.ir., Zafarmand-Samarin M; Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran. Electronic address: m.zafarmand@ihu.ac.ir., Sadeghi D; Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran. Electronic address: Kpsadeghi@ihu.ac.ir., Fathi J; Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran., Samiei-Abianeh H; Department of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran; Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Jazyk: angličtina
Zdroj: Molecular immunology [Mol Immunol] 2024 Sep; Vol. 173, pp. 53-60. Date of Electronic Publication: 2024 Jul 24.
DOI: 10.1016/j.molimm.2024.07.002
Abstrakt: Introduction: Shigellosis is a gastrointestinal disease causes high morbidity and mortality worldwide, however, there is no anti-Shigella vaccine. The use of antibiotics in shigellosis treatment exacerbates antibiotic resistance. Antibodies, particularly egg yolk antibody (IgY), offer a promising approach to address this challenge. This study aimed to investigate the prophylactic effect of IgY produced against a recombinant chimeric protein containing the immunogens IpaD, IpaB, StxB, and VirG from Shigella.
Methods: The chimeric protein, comprising IpaD, IpaB, StxB, and VirG, was expressed in E. coli BL21 and purified using the Ni-NTA column. Following immunization of chickens, IgY was extracted from egg yolk using the PEG-6000 method and analyzed through SDS-PAGE and ELISA techniques. Subsequently, the prophylactic efficacy of IgY was assessed by challenging of mice with 10 LD50 of S. dysenteriae and administering different concentrations of IgY (1.25, 2.5, 5, and 10 mg/kg) under various time conditions.
Results: The recombinant protein, weighing 82 kDa, was purified and confirmed by western blotting. The IgY concentration was determined as 9.5 mg/ml of egg yolk and the purity of the extracted IgY was over 90 %. The results of the ELISA showed that at least 19 ng of pure antibody identified recombinant protein and reacts with it. The challenge test employing IgY and Shigella demonstrated a direct correlation between the survival rate and antibody concentration, with increased concentrations leading to decreased mortality rates. Treatment of mice with 10 mg/kg IgY leads to 80 % survival of the mice against 10 LD50 S. dysenteriae.
Conclusion: Our findings suggest that IgY may offer therapeutic potential in treating Shigella infections and combating antibiotic resistance.
Competing Interests: Declaration of Competing Interest We declare that we have no conflict of interest.
(Copyright © 2024 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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