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

Yıl 2020, , 540 - 544, 01.07.2020

Mohammad Kazem Sharifi Yazdi , Seyed Davar Siadat Mohammad Khalifeh-gholi Sara Sharifi-yazdi Ahmad Fayaz-bakhsh Mohammad Saleh Safari

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

Cited By: 1

Öz

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.

Anahtar Kelimeler

External mucous vesicles, Brucella aborotus, Brucellosis vaccine

Destekleyen Kurum

Tehran university of Medical Sciences

Proje Numarası

26355

Teşekkür

thank you for considering my article

Bruselloz aşısı için aday olarak Brucella abortus'un dış zar veziküllerinin ekstraksiyonu ve biyolojik değerlendirmesi

Öz

Amaç: Bruselloz, hayvan ve insan sağlığını etkileyen endemik bir zoonotik hastalıktır. Son birkaç on yılda, hastalığı kontrol etmek için daha güvenli Brucella aşıları geliştirmek için çok araştırma yapılmıştır. Dış zar vezikülleri (OMV'ler), insan brusellozunu önlemek için bir alt birim aşı için immünojenik yapılar olarak düşünülmüştür. Bu çalışmanın amacı Brusella abortus'un dış zar veziküllerini bruselloz aşısı için aday olarak değerlendirmektir.
Yöntemler: Bu çalışmada Brucella abortus, S99 suşu kullanıldı. Brucella abortusun kitle kültivasyonundan sonra OMV ekstraksiyonu, Ultracentrifugation ve Sodium Dosoxycotinate tarafından gerçekleştirildi. SDS-PAGE yöntemi, fizikokimyasal özellikleri değerlendirmek için protein paternini ve elektron mikroskopisini gözlemlemek için kullanıldı. OMV'deki LOS miktarı da LAL testi ile ölçüldü. Protein konsantrasyonunun belirlenmesinden sonra farelere belirli bir konsantrasyonda enjekte edildi ve antikoru değerlendirmek için kan örnekleri alındı. Son olarak serum izolasyonundan sonra antikor üretimi ELISA yöntemi ile ölçüldü.
Bulgular: OMV'de mevcut protein miktarı 0,1 mg/ml idi. OMV'nin boyutu 55 ila 150 nm idi ve SDS-PAGE değerlendirmesinde protein aralığı >25 kDa idi. LAL testindeki LOS değeri yetkili aralıkta rapor edildi. İlk enjeksiyondan sonra IgG düzeylerinde (P=0,001) kontrol grubuna kıyasla ve daha sonraki güçlendiricilerde anlamlı artış gözlendi. En yüksek tepki oranı ikinci güçlendiricide gözlenmiştir.
Sonuç: OMV ekstraksiyon sırasında uzamsal şeklini korumaktadır ve Brucella düşüklerine karşı yüksek derecede spesifik antikorların üretimini indükleme potansiyeline sahiptir. Bu nedenle klinik fazları inceledikten sonra aday bir aşı olarak düşünülebilir.

Anahtar Kelimeler

Dış mukoza vezikülleri, Brucella abortus S99, Bruselloz aşısı

Proje Numarası

26355

Kaynakça

• 1. Pappas G, Papadimitriou P, Akritidis N, Christou L, Tsianos EV. The new global map of human brucellosis. Lancet Infectious Diseases. 2006; 6(2):91–9.
• 2. Nicoletti P. Vaccination. In: Nielsen K, Duncan JR, editors. Animal Brucellosis. Boca Raton, Fla, USA: CRC Press; 1990. pp. 284–299. `
• 3. Schurig GG, Sriranganathan N, Corbel MJ. Brucellosis vaccines: past, present and future. Veterinary Microbiology. 2002;90(1–4):479–96.
• 4. Montaraz JA, Winter AJ. Comparison of living and nonliving vaccines for Brucella abortus in BALB/c mice. Infection and Immunity. 1986;53(2):245–51.
• 5. Winter AJ, Rowe GE. Comparative immune responses to native cell envelope antigens and the hot sodium dodecyl sulfate insoluble fraction (PG) of Brucella abortus in cattle and mice. Veterinary Immunology and Immunopathology. 1988;18(2):49–63.
• 6. Olsen SC, Stoffregen WS. Essential role of vaccines in brucellosis control and eradication programs for livestock. Expert Review of Vaccines. 2005;4(6):915–28.
• 7. Marzetti S, Carranza C, Roncallo M, Escobar GI, Lucero NE. Recent trends in humanBrucellacanisinfection. Comparative Immunology, Microbiology and Infectious Disease. 2013;36(1):55–61.
• 8. Perkins SD, Smither SJ, Atkins HS. Towards a Brucella vaccine for humans. FEMS Microbiology Reviews. 2010;34(3):379–94.
• 9. Pasquevich KA, Ibañez AE, Coria LM, et al. An oral vaccine based on U-Omp19 induces protection against B. abortus mucosal challenge by inducing an adaptive IL-17 immune response in mice. PLoS ONE. 2011; 6(1):e16203.
• 10. Haghi F, Najar Peerayeha S, Siadat SD, Zeighami H. Recombinant outer membrane secretin PilQ406–770 as a vaccine candidate for serogroup B Neisseria meningitidis. Vaccine.2012;30(9):1710-4.
• 11. Moshiri A, Dashtbani Roozbehani A, Peerayeh S, Siadat SD. Outer membrane vesicle: a macromolecule with multifunctional activity. Hum Vaccin Immunother. 2012;8(7):953-5.
• 12. Delbaz SA, Siadat SD, Aghasadeghi MR, Piryaie M, Najar Peerayeh S, Mousavi SF et al. Biological and Immunological Evaluation of Neisseria meningitidis Serogroup A Outer Membrane Vesicle as Vaccine Candidates. Jundishapur J Microbiol.2013;6(4):e5007.
• 13. Dorneles EMS, Lima GK, Teixeira-Carvalho A, Araújo MS, Martins-Filho OA, Sriranganathan N, et al. Immune Response of Calves Vaccinated with Brucella abortusS19 or RB51 and Revaccinated RB51. PLOS ONE 2015;10(9):e0136696. https://doi.org/10.1371/journal.pone.0136696.
• 14. Dorneles EMS, Sriranganathan N, Lage AP. Recent advances in Brucella abortus vaccines. Vet Res. 2015;46(1):76.
• 15. Avila-Calderón ED, Ahidé Lopez M, Sriranganathan N, Boyle SM, Contreras-Rodriguez A. A History of the Development of Brucella Vaccines. BioMed Research International.Volume. 2013;743509.
• 16. Kaur G, Singh S, Sunil Kumar BV, Mahajan K, Verma R. Characterization and Immunogenicity of Outer Membrane Vesicles from Brucella abortus. J Immunoassay Immunochem. 2016;37(3):261-72. doi: 10.1080/15321819.2015.1132231.
• 17. Pollak CN, Delpino MV, Fossati CA, Baladi PC. Outer Membrane Vesicles from Brucella abortus Promote Bacterial Internalization by Human Monocytes and Modulate Their Innate Immune Response. PloS ONE 7(11):e50214.
• 18. Sharifi-yazdi MK, Beheahtnejad AH, Hedayatpour A, Sharifi-yazdi S, Mehrani F. Comparative study of Mycobacterium tuberculosis and Mycobacterium bovis protein profiles. J surgery Medicine. 2018;2(3):223–7.