Examination of some biological properties of glycoprotein subunits of PHY-LMV.42 strain of Newcastle disease virus
Ispitivanje nekih bioloških karakteristika glikoproteinskih subjedinica soja PHY-LMV.42 virusa Newcastle bolesti živine
2015
Authors
Milić, NenadNišavić, Jakov
Borozan, Sunčica
Zorić, Andrea
Lazić, Sava
Petrović, Tamaš
Rašić, Zoran
Article (Published version)
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The objective of our work was to investigate some biological characteristics of purified glycoprotein subunits of Newcastle disease virus strain PHY-LMV.42 isolated from pigeons for the purpose of vaccine production. PHY-LMV.42 strain of Newcastle disease virus was multiplied by successive passages in embryonated eggs and identified by the methods of Reverse transcriptase PCR and Real-Time PCR along with F gene sequencing. Proving the presence of HN and F antigene in the virus subunits samples was carried out by hemagglutination inhibition method with referent immune sera. Biochemical characterization of glycoprotein subunits was performed by SDS-PAGE method as well as liquid chromatography with mass spectrometry (LC ESI-TOF-MS/MS). Testing for the virus subunits immunogenicity was carried out in biological experiment on 75 laying hens Tetra-SSL and 25 chickens Isa Brown by inducing an artificial infection with Hertz 33 strain of the virus. Low concentrations of the virus antigens of 0....36 mg/ml along with glycoprotein fractions of 77 i 58 kDa manifested a strong hemagglutination activity of 4096 HJ/0,1ml. The subunit vaccines of 256 and 128 HJ/0.5 ml induced a protective immune response in all the vaccinated animals. Based on the obtained results it can be concluded that low concentrations of purified virus subunits of PHY-LMV.42 strain can be used for preparing of effective vaccines.
Cilj našeg istraživanja je bilo ispitivanje bioloških karakteristika prečišćenih glikoproteinskih subjedinica soja PHY-LMV.42 virusa Newcastle bolesti izolovanog iz golubova radi njihovog korišćenja za pripremanje vakcine. Soj PHY-LMV.42 virusa Newcastle bolesti je umnožavan sukcesivnim pasažama u kokošijim embrionima i identifikovan metodama Reverse transcriptase PCR i Real- Time PCR uz sekvenciranje F gena. Dokazivanje prisustva HN i F antigena u uzorcima virusnih subjedinica vršeno je metodom inhibicije hemaglutinacije sa referentnim imunim serumima. Biohemijska karakterizacija glikoproteinskih subjedinica izvršena je primenom metoda SDS-PAGE i tečne hromatografije sa masenom spektrometrijom (LC ESI-TOF-MS/MS). Ispitivanje imunogenosti virusnih subjedinica sprovedeno je u biološkom ogledu na ukupno 75 kokoši nosilja Tetra-SSL i 25 pilića Isa Brown uz izvođenje veštačke infekcije sojem Hertz 33 navedenog virusa. Niske koncentracije virusnih antigena od 0,36 mg/ml sa glikoproteinskim ...frakcijama od 77 i 58 kDa su ispoljavale snažnu hemaglutinacionu aktivnost od 4096 HJ/0,1ml. Subjedinične vakcine od 256 i 128 HJ/0,5 ml, indukovale su imunološki odgovor zaštitnog karaktera kod svih vakcinisanih životinja. Na osnovu dobijenih rezultata može se zaključiti da se niske koncentracije prečišćenih virusnih subjedinica soja PHY-LMV.42 mogu koristiti za pripremanje efikasne vakcine. PR Projekat Ministarstva nauke Republike Srbije, br. TR 31008.
Keywords:
Newcastle disease virus / glycoprotein subunits / SDS-PAGE / LC ESI-TOF-MS/MS / immunization / virus Newcastle bolesti / glikoproteinske subjedinice / SDS-PAGE / LC ESI-TOF-MS/MS / imunizacijaSource:
Veterinarski Glasnik, 2015, 69, 5-6, 337-355Publisher:
- Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
Funding / projects:
- Development and application of molecular methods based on polymerase chain reaction (PCR) in rapid and direct identification of Newcastle disease virus strains and examination of immunogenicity of subunit vaccine prepared from their antigens (RS-MESTD-Technological Development (TD or TR)-31008)
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Institution/Community
Fakultet veterinarske medicineTY - JOUR AU - Milić, Nenad AU - Nišavić, Jakov AU - Borozan, Sunčica AU - Zorić, Andrea AU - Lazić, Sava AU - Petrović, Tamaš AU - Rašić, Zoran PY - 2015 UR - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1221 AB - The objective of our work was to investigate some biological characteristics of purified glycoprotein subunits of Newcastle disease virus strain PHY-LMV.42 isolated from pigeons for the purpose of vaccine production. PHY-LMV.42 strain of Newcastle disease virus was multiplied by successive passages in embryonated eggs and identified by the methods of Reverse transcriptase PCR and Real-Time PCR along with F gene sequencing. Proving the presence of HN and F antigene in the virus subunits samples was carried out by hemagglutination inhibition method with referent immune sera. Biochemical characterization of glycoprotein subunits was performed by SDS-PAGE method as well as liquid chromatography with mass spectrometry (LC ESI-TOF-MS/MS). Testing for the virus subunits immunogenicity was carried out in biological experiment on 75 laying hens Tetra-SSL and 25 chickens Isa Brown by inducing an artificial infection with Hertz 33 strain of the virus. Low concentrations of the virus antigens of 0.36 mg/ml along with glycoprotein fractions of 77 i 58 kDa manifested a strong hemagglutination activity of 4096 HJ/0,1ml. The subunit vaccines of 256 and 128 HJ/0.5 ml induced a protective immune response in all the vaccinated animals. Based on the obtained results it can be concluded that low concentrations of purified virus subunits of PHY-LMV.42 strain can be used for preparing of effective vaccines. AB - Cilj našeg istraživanja je bilo ispitivanje bioloških karakteristika prečišćenih glikoproteinskih subjedinica soja PHY-LMV.42 virusa Newcastle bolesti izolovanog iz golubova radi njihovog korišćenja za pripremanje vakcine. Soj PHY-LMV.42 virusa Newcastle bolesti je umnožavan sukcesivnim pasažama u kokošijim embrionima i identifikovan metodama Reverse transcriptase PCR i Real- Time PCR uz sekvenciranje F gena. Dokazivanje prisustva HN i F antigena u uzorcima virusnih subjedinica vršeno je metodom inhibicije hemaglutinacije sa referentnim imunim serumima. Biohemijska karakterizacija glikoproteinskih subjedinica izvršena je primenom metoda SDS-PAGE i tečne hromatografije sa masenom spektrometrijom (LC ESI-TOF-MS/MS). Ispitivanje imunogenosti virusnih subjedinica sprovedeno je u biološkom ogledu na ukupno 75 kokoši nosilja Tetra-SSL i 25 pilića Isa Brown uz izvođenje veštačke infekcije sojem Hertz 33 navedenog virusa. Niske koncentracije virusnih antigena od 0,36 mg/ml sa glikoproteinskim frakcijama od 77 i 58 kDa su ispoljavale snažnu hemaglutinacionu aktivnost od 4096 HJ/0,1ml. Subjedinične vakcine od 256 i 128 HJ/0,5 ml, indukovale su imunološki odgovor zaštitnog karaktera kod svih vakcinisanih životinja. Na osnovu dobijenih rezultata može se zaključiti da se niske koncentracije prečišćenih virusnih subjedinica soja PHY-LMV.42 mogu koristiti za pripremanje efikasne vakcine. PR Projekat Ministarstva nauke Republike Srbije, br. TR 31008. PB - Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd T2 - Veterinarski Glasnik T1 - Examination of some biological properties of glycoprotein subunits of PHY-LMV.42 strain of Newcastle disease virus T1 - Ispitivanje nekih bioloških karakteristika glikoproteinskih subjedinica soja PHY-LMV.42 virusa Newcastle bolesti živine VL - 69 IS - 5-6 SP - 337 EP - 355 DO - 10.2298/VETGL1506337M ER -
@article{ author = "Milić, Nenad and Nišavić, Jakov and Borozan, Sunčica and Zorić, Andrea and Lazić, Sava and Petrović, Tamaš and Rašić, Zoran", year = "2015", abstract = "The objective of our work was to investigate some biological characteristics of purified glycoprotein subunits of Newcastle disease virus strain PHY-LMV.42 isolated from pigeons for the purpose of vaccine production. PHY-LMV.42 strain of Newcastle disease virus was multiplied by successive passages in embryonated eggs and identified by the methods of Reverse transcriptase PCR and Real-Time PCR along with F gene sequencing. Proving the presence of HN and F antigene in the virus subunits samples was carried out by hemagglutination inhibition method with referent immune sera. Biochemical characterization of glycoprotein subunits was performed by SDS-PAGE method as well as liquid chromatography with mass spectrometry (LC ESI-TOF-MS/MS). Testing for the virus subunits immunogenicity was carried out in biological experiment on 75 laying hens Tetra-SSL and 25 chickens Isa Brown by inducing an artificial infection with Hertz 33 strain of the virus. Low concentrations of the virus antigens of 0.36 mg/ml along with glycoprotein fractions of 77 i 58 kDa manifested a strong hemagglutination activity of 4096 HJ/0,1ml. The subunit vaccines of 256 and 128 HJ/0.5 ml induced a protective immune response in all the vaccinated animals. Based on the obtained results it can be concluded that low concentrations of purified virus subunits of PHY-LMV.42 strain can be used for preparing of effective vaccines., Cilj našeg istraživanja je bilo ispitivanje bioloških karakteristika prečišćenih glikoproteinskih subjedinica soja PHY-LMV.42 virusa Newcastle bolesti izolovanog iz golubova radi njihovog korišćenja za pripremanje vakcine. Soj PHY-LMV.42 virusa Newcastle bolesti je umnožavan sukcesivnim pasažama u kokošijim embrionima i identifikovan metodama Reverse transcriptase PCR i Real- Time PCR uz sekvenciranje F gena. Dokazivanje prisustva HN i F antigena u uzorcima virusnih subjedinica vršeno je metodom inhibicije hemaglutinacije sa referentnim imunim serumima. Biohemijska karakterizacija glikoproteinskih subjedinica izvršena je primenom metoda SDS-PAGE i tečne hromatografije sa masenom spektrometrijom (LC ESI-TOF-MS/MS). Ispitivanje imunogenosti virusnih subjedinica sprovedeno je u biološkom ogledu na ukupno 75 kokoši nosilja Tetra-SSL i 25 pilića Isa Brown uz izvođenje veštačke infekcije sojem Hertz 33 navedenog virusa. Niske koncentracije virusnih antigena od 0,36 mg/ml sa glikoproteinskim frakcijama od 77 i 58 kDa su ispoljavale snažnu hemaglutinacionu aktivnost od 4096 HJ/0,1ml. Subjedinične vakcine od 256 i 128 HJ/0,5 ml, indukovale su imunološki odgovor zaštitnog karaktera kod svih vakcinisanih životinja. Na osnovu dobijenih rezultata može se zaključiti da se niske koncentracije prečišćenih virusnih subjedinica soja PHY-LMV.42 mogu koristiti za pripremanje efikasne vakcine. PR Projekat Ministarstva nauke Republike Srbije, br. TR 31008.", publisher = "Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd", journal = "Veterinarski Glasnik", title = "Examination of some biological properties of glycoprotein subunits of PHY-LMV.42 strain of Newcastle disease virus, Ispitivanje nekih bioloških karakteristika glikoproteinskih subjedinica soja PHY-LMV.42 virusa Newcastle bolesti živine", volume = "69", number = "5-6", pages = "337-355", doi = "10.2298/VETGL1506337M" }
Milić, N., Nišavić, J., Borozan, S., Zorić, A., Lazić, S., Petrović, T.,& Rašić, Z.. (2015). Examination of some biological properties of glycoprotein subunits of PHY-LMV.42 strain of Newcastle disease virus. in Veterinarski Glasnik Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd., 69(5-6), 337-355. https://doi.org/10.2298/VETGL1506337M
Milić N, Nišavić J, Borozan S, Zorić A, Lazić S, Petrović T, Rašić Z. Examination of some biological properties of glycoprotein subunits of PHY-LMV.42 strain of Newcastle disease virus. in Veterinarski Glasnik. 2015;69(5-6):337-355. doi:10.2298/VETGL1506337M .
Milić, Nenad, Nišavić, Jakov, Borozan, Sunčica, Zorić, Andrea, Lazić, Sava, Petrović, Tamaš, Rašić, Zoran, "Examination of some biological properties of glycoprotein subunits of PHY-LMV.42 strain of Newcastle disease virus" in Veterinarski Glasnik, 69, no. 5-6 (2015):337-355, https://doi.org/10.2298/VETGL1506337M . .