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
Аутори
Milić, NenadNišavić, Jakov
Borozan, Sunčica
Zorić, Andrea
Lazić, Sava
Petrović, Tamaš
Rašić, Zoran
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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.
Кључне речи:
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 / imunizacijaИзвор:
Veterinarski Glasnik, 2015, 69, 5-6, 337-355Издавач:
- Univerzitet u Beogradu - Fakultet veterinarske medicine, Beograd
Финансирање / пројекти:
- Развој и примена молекуларних метода заснованих на ланчаној реакцији полимеразе (PCR) у брзој и директној идентификацији сојева вируса Неwцастле болести живине и испитивање имуногености субјединичне вакцине припремљене од њихових антигена (RS-MESTD-Technological Development (TD or TR)-31008)
Колекције
Институција/група
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 . .