Extraction and biological evaluation of external membrane vesicles of Brucella abortus as a candidate for brucellosis vaccine

Mohammad Kazem Sharifi Yazdi; Seyed Davar Siadat; Mohammad Khalifeh-Gholi; Sara Sharifi-Yazdi; Ahmad Fayaz-Bakhsh; Mohammad Saleh Safari

doi:10.28982/josam.674662

Authors

Mohammad Kazem Sharifi Yazdi https://orcid.org/0000-0001-9060-5908
Seyed Davar Siadat Zoonosis Research Centre, Tehran University of Medical Sciences, Tehran, Iran
Mohammad Khalifeh-Gholi
Sara Sharifi-Yazdi MD student, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Ahmad Fayaz-Bakhsh School of Public Health and Health Information Research Center, Tehran University of Medical Sciences, Tehran, Iran
Mohammad Saleh Safari Veterinary Medicine student, Faculty of Science and Research Branch, Islamic Azad University, Tehran, Iran

DOI:

https://doi.org/10.28982/josam.674662

Keywords:

External mucous vesicles, Brucella aborotus, Brucellosis vaccine

Abstract

Aim: Brucellosis is an endemic zoonotic disease affecting animal and human health. In the last several decades, much research has been performed to develop safer Brucella vaccines to control the disease. Outer membrane vesicles (OMVs) have been considered as immunogenic structures for a subunit vaccine to prevent human brucellosis. The aim of this study was to evaluate the external membrane vesicles of Brucella abortus as a candidate for brucellosis vaccine.
Methods: In this study Brucella abortus, S99 strain was used. Extraction of OMV after mass cultivation of Brucella abortus was performed by Ultracentrifugation and Sodium Dosoxycotinate. The SDS-PAGE method was used to observe the protein pattern and electron microscopy, to evaluate the physicochemical properties. The amount of LOS in OMV was also measured by LAL test. After determination of the protein concentration, mice were injected at a specific concentration and blood samples were obtained to evaluate the antibody. Finally, after serum isolation, the antibody production was measured by the ELISA method.
Results: The amount of protein present in OMV was 0.1 mg/ml. The size of OMV was 55 to 150 nm, and in the SDS-PAGE assessment, the protein range was >25 kDa. The LOS value in the LAL test was reported within the authorized range. Significant increase in IgG levels was observed after the first injection (P=0.001) compared to the control group, and in subsequent boosters as well. The highest response rate was observed in the second booster.
Conclusion: OMV maintains its spatial shape during extraction and has the potential to induce the production of a high degree of specific antibodies against Brucella abortus. For this reason, it can be considered as a candidate vaccine after examining clinical phases.

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