Subgroup J Avian leucosis virus (ALV-J) strain NX0101 was inoculated into chicken embryo fibroblasts (CEF) monolayers in 6-well plates. The six wells of CEF inoculated with NX0101 were divided into groups A (without a...Subgroup J Avian leucosis virus (ALV-J) strain NX0101 was inoculated into chicken embryo fibroblasts (CEF) monolayers in 6-well plates. The six wells of CEF inoculated with NX0101 were divided into groups A (without anti-ALV-J serum in the medium); B (with anti-ALV-J serum in the medium), then viruses from each well of both groups were separately passed in CEF every 6 d; formed their independent passage lineages. For each lineage of both groups, gp85 genes of the viruses in the 10th, 20th; 30th passages were amplified, cloned; sequenced. The sequence data indicated that the homologies of gp85 at aa level between the primary virus; the passed viruses of different passages of 3 lineages in group A were 97.7%–99.7%;; the homologies of gp85 between the primary virus; the passed viruses of different passages of 3 lineages in group B were 93.8%–96.1%. Analysis of the ratios of nonsynonium (NS) vs synonium (S) mutations of nucleic acids demonstrated that NS/S in 3 highly variable (hr-) regions at aa#110–120, aa#141–151; aa#189–194 of gp85 in 3 lineages of group A were 2 (8/4), 1(3/3); 1.3 (4/3), however, NS/S in the same 3 hr-regions of group B were 4.1 (13/3), 4.7 (14/3); 3.3 (11/3). This study is the first demonstration of influence of immune selective pressure on evolution of ALV-J gp85 by specific antibodies under the controlled in vitro experiments.展开更多
[ Objective] The paper was to understand the natural infection status of avian leukemia in some indigenous chicken breeds of China. [ Method ] Using ELISA assay and virus isolation method, epidemiological investigatio...[ Objective] The paper was to understand the natural infection status of avian leukemia in some indigenous chicken breeds of China. [ Method ] Using ELISA assay and virus isolation method, epidemiological investigation of ALV-AB and ALV-J avian leukemia of 10 indigenous chicken breeds were conducted. ALV dynamics were monitored in F2 generation of four chicken lines. ALV pollution of attenuated live vaccines used in raising process was also inspected through sampling method. [ Result] The positive rate of ALV-P27 antigen was 0 -62.1% ; the positive rate of ALV-AB antibody was 0 -25.0% ; the positive rate of ALV-J antibody was 0 - 59.0% ; the positive rate of virus isolation was 0 - 22.0%. The positive rate of ALV-P＇27 antigen in 1 -day-old chick mecunium of four lines was 6.0% - 67.0%. The positive rate of virus isolation in 6-week-old chickens was 2.0% - 34.3%. Two kinds of vaccines from two hatches pro- duced by a manufacturer were polluted by ALV. [ Conclusion] Most indigenous chickens were infected by ALV. There were great differences among different breeds of indigenous chicken, which might be related to ALV genetic resistance of different indigenous chickens. The ALV positive rates of F： chicks were slightly enhanced in some lines, which might be related to vaccine pollution.展开更多
文摘Subgroup J Avian leucosis virus (ALV-J) strain NX0101 was inoculated into chicken embryo fibroblasts (CEF) monolayers in 6-well plates. The six wells of CEF inoculated with NX0101 were divided into groups A (without anti-ALV-J serum in the medium); B (with anti-ALV-J serum in the medium), then viruses from each well of both groups were separately passed in CEF every 6 d; formed their independent passage lineages. For each lineage of both groups, gp85 genes of the viruses in the 10th, 20th; 30th passages were amplified, cloned; sequenced. The sequence data indicated that the homologies of gp85 at aa level between the primary virus; the passed viruses of different passages of 3 lineages in group A were 97.7%–99.7%;; the homologies of gp85 between the primary virus; the passed viruses of different passages of 3 lineages in group B were 93.8%–96.1%. Analysis of the ratios of nonsynonium (NS) vs synonium (S) mutations of nucleic acids demonstrated that NS/S in 3 highly variable (hr-) regions at aa#110–120, aa#141–151; aa#189–194 of gp85 in 3 lineages of group A were 2 (8/4), 1(3/3); 1.3 (4/3), however, NS/S in the same 3 hr-regions of group B were 4.1 (13/3), 4.7 (14/3); 3.3 (11/3). This study is the first demonstration of influence of immune selective pressure on evolution of ALV-J gp85 by specific antibodies under the controlled in vitro experiments.
基金Supported by Special Fund for Agro-scientific Research in the Public Interest(201203055)Agricultural Three-new Engineering Project of Jiangsu Province(SXGC[2014]288)+1 种基金Natural Science Foundation of Jiangsu Province(20151317)Science and Technology Project of Yangzhou City(YZ2014144)
文摘[ Objective] The paper was to understand the natural infection status of avian leukemia in some indigenous chicken breeds of China. [ Method ] Using ELISA assay and virus isolation method, epidemiological investigation of ALV-AB and ALV-J avian leukemia of 10 indigenous chicken breeds were conducted. ALV dynamics were monitored in F2 generation of four chicken lines. ALV pollution of attenuated live vaccines used in raising process was also inspected through sampling method. [ Result] The positive rate of ALV-P27 antigen was 0 -62.1% ; the positive rate of ALV-AB antibody was 0 -25.0% ; the positive rate of ALV-J antibody was 0 - 59.0% ; the positive rate of virus isolation was 0 - 22.0%. The positive rate of ALV-P＇27 antigen in 1 -day-old chick mecunium of four lines was 6.0% - 67.0%. The positive rate of virus isolation in 6-week-old chickens was 2.0% - 34.3%. Two kinds of vaccines from two hatches pro- duced by a manufacturer were polluted by ALV. [ Conclusion] Most indigenous chickens were infected by ALV. There were great differences among different breeds of indigenous chicken, which might be related to ALV genetic resistance of different indigenous chickens. The ALV positive rates of F： chicks were slightly enhanced in some lines, which might be related to vaccine pollution.
