Hydro-Pericardium Syndrome (HPS) is viral problem of commercial poultry caused by aviadeno virus type-4. In Pakistan the problems have been controlled by administering inactivated infected liver homogenate vaccine (IL...Hydro-Pericardium Syndrome (HPS) is viral problem of commercial poultry caused by aviadeno virus type-4. In Pakistan the problems have been controlled by administering inactivated infected liver homogenate vaccine (ILHV). The use of liver based HPS vaccines remained potential threat for having hypersensitivity reactions in poultry. The current study was carried out to compare the serological potency of HPS ILHV to vero cell line adopted vaccine in term of anti HPS-ELISA antibody titers. 14 HPS virus vaccines were prepared based on different concentration of antigen, type of adjuvants and source of virus substrate. Total of 160 birds were divided into 16 groups each containing 10 birds. At day of 14th age each bird of every group was injected with 0.3 ml dose of respective vaccine. It was observed that HPS infected liver based vaccine having 1 × 105.6, 1 × 105.6 and 1 × 103.6 bird lethal dose 50 induced 1092.10, 875.25 and 702.2 anti-HPS ELISA antibody titer respectively. The 20, 25 and 30 doses/gm HPS infected liver vaccine induced 110.4, 1071.9 and 1037.8 anti-HPS ELISA antibody titer respectively. Montanide based tissue culture HPS vaccine showed significantly higher 1148.45 anti-HPS ELISA antibody titer to aluminium hydroxide based vaccine (137.2) (P 5.6 TCID50 is serological potent against field infection. The vaccines based on such formulation could be prepared in future for effective immuno-prophylaxis against HPS virus.展开更多
The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate ...The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 - 12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 10<sup>7</sup> FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 - 4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study in Türkiye that used microcarriers.展开更多
文摘Hydro-Pericardium Syndrome (HPS) is viral problem of commercial poultry caused by aviadeno virus type-4. In Pakistan the problems have been controlled by administering inactivated infected liver homogenate vaccine (ILHV). The use of liver based HPS vaccines remained potential threat for having hypersensitivity reactions in poultry. The current study was carried out to compare the serological potency of HPS ILHV to vero cell line adopted vaccine in term of anti HPS-ELISA antibody titers. 14 HPS virus vaccines were prepared based on different concentration of antigen, type of adjuvants and source of virus substrate. Total of 160 birds were divided into 16 groups each containing 10 birds. At day of 14th age each bird of every group was injected with 0.3 ml dose of respective vaccine. It was observed that HPS infected liver based vaccine having 1 × 105.6, 1 × 105.6 and 1 × 103.6 bird lethal dose 50 induced 1092.10, 875.25 and 702.2 anti-HPS ELISA antibody titer respectively. The 20, 25 and 30 doses/gm HPS infected liver vaccine induced 110.4, 1071.9 and 1037.8 anti-HPS ELISA antibody titer respectively. Montanide based tissue culture HPS vaccine showed significantly higher 1148.45 anti-HPS ELISA antibody titer to aluminium hydroxide based vaccine (137.2) (P 5.6 TCID50 is serological potent against field infection. The vaccines based on such formulation could be prepared in future for effective immuno-prophylaxis against HPS virus.
文摘The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 - 12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 10<sup>7</sup> FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 - 4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study in Türkiye that used microcarriers.
