鸡传染性贫血病病毒在MDCC-MSB1细胞的传代致弱研究
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摘要
为研究鸡传染性贫血病(Chicken infectious anemia,CIA)减毒活疫苗,试验将鸡传染性贫血病病毒(CIAV)在鸡淋巴细胞MDCC-MSB1上进行37℃和40℃传代致弱。结果显示:40℃条件下传至60代和37℃下传至90代,病毒的VP1氨基酸分别有2个和3个突变,两种条件下有2个突变是在同一位置;VP2和VP3很稳定,在试验中未发生氨基酸突变;致病性试验发现,突变毒株仍具有致病性,但有降低的趋势。试验表明,鸡淋巴细胞传代CIAV可以引发包含中和抗原表位的VP1突变,且提高温度能促进变异速度,为进一步研究CIAV的变异特征、弱毒疫苗研发提供了参考依据。
To study attenuated live vaccine for chicken infectious anemia(CIA), chicken anemia virus(CIAV) was passaged in MDCC-MSB1 cell at 37 ℃ and 40 ℃, respectively. The results showed that three amino acid mutations occurred in VP1 protein encoded during 37 ℃, including two amino acid mutations occurred in VP1 during 40 ℃;There were no amino acid mutations in this experiment in VP2 and VP3 protein encoded; In vivo assays, the mutant strains remained pathogenicity but they had a tendency to attenuate. It′s suggested that MDCC-MSB1 could trigger amino acid mutations in VP1 protein, which contains the neutralizing antigen epitopes of CIAV. Moreover, increasing temperature could promote the mutation rate. All these data provided a theoretical basis for studying CIAV variation characteristics and CIA attenuated vaccine.
To study attenuated live vaccine for chicken infectious anemia(CIA), chicken anemia virus(CIAV) was passaged in MDCC-MSB1 cell at 37 ℃ and 40 ℃, respectively. The results showed that three amino acid mutations occurred in VP1 protein encoded during 37 ℃, including two amino acid mutations occurred in VP1 during 40 ℃;There were no amino acid mutations in this experiment in VP2 and VP3 protein encoded; In vivo assays, the mutant strains remained pathogenicity but they had a tendency to attenuate. It′s suggested that MDCC-MSB1 could trigger amino acid mutations in VP1 protein, which contains the neutralizing antigen epitopes of CIAV. Moreover, increasing temperature could promote the mutation rate. All these data provided a theoretical basis for studying CIAV variation characteristics and CIA attenuated vaccine.
引文
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[8]肖庆利,张志芳,何家禄.鸡贫血病毒VP1和VP2蛋白在家蚕中的联合表达[J].生物工程学报,2001(3):283-287.
[9]MOEINI H,RAHIM R A,OMAR A R,et al.Lactobacillus acidophilus as a live vehicle for oral immunization against chicken anemia virus[J].Appl Microbiol Biotechnol,2011,90(1):77-88.
[10]蔺文成,张新珩,李昕建,等.4株鸡传染性贫血病毒的分离鉴定及其遗传进化分析[J].中国预防兽医学报,2016(9):695-699.
[11]GORYO M,SUWA T,MATSUMOTO S,et al.Serial propagation and purification of chicken anaemia agent in MDCCMSB1 cell line[J].Avian Pathol,1987,16(1):149-163.
[12]YAMAGUCHI S,IMADA T,KAJI N,et al.Identification of a genetic determinant of pathogenicity in chicken anaemia virus[J].J Gen Virol,2001,82(5):1233-1238.
[13]TODD D,CONNOR T J,CALVERT V M,et al.Molecular cloning of an attenuated chicken anaemia virus isolate following repeated cell culture passage[J].Avian Pathol,1995,24(1):171-187.
[14]MEEHAN B M,TODD D,CREELAN J L,et al.Investigation of the attenuation exhibited by a molecularly cloned chicken anemia virus isolate by utilizing a chimeric virus approach[J].J Virol,1997,71(11):8362-8367.
[2]GORYO M,SUGIMURA H,MATSUMOTO S,et al.Isolation of an agent inducing chicken anaemia[J].Avian Pathol,1985,14(4):483-496.
[3]NOTEBORN M H,DE BOER G F,VAN ROOZELAAR D J,et al.Characterization of cloned chicken anemia virus DNA that contains all elements for the infectious replication cycle[J].J Virol,1991,65(6):3131-3139.
[4]崔现兰,幸桂香,吴东来,等.鸡传染性贫血病病毒的鉴定[J].中国畜禽传染病,1992(6):3-5.
[5]王莉莉,艾武,吴荫伟,等.山东省鸡传染性贫血流行病学调查[J].山东家禽,2002(8):8-9.
[6]PAGES-MANTE A,SAUBI N,ARTIGAS C,et al.Experimental evaluation of an inactivated vaccine against chicken anaemia virus[J].Avian Pathol,1997,26(4):721-729.
[7]王秀荣,王笑梅,杨林,等.鸡传染性贫血病灭活苗的研制[J].中国预防兽医学报,1999(1):20-22.
[8]肖庆利,张志芳,何家禄.鸡贫血病毒VP1和VP2蛋白在家蚕中的联合表达[J].生物工程学报,2001(3):283-287.
[9]MOEINI H,RAHIM R A,OMAR A R,et al.Lactobacillus acidophilus as a live vehicle for oral immunization against chicken anemia virus[J].Appl Microbiol Biotechnol,2011,90(1):77-88.
[10]蔺文成,张新珩,李昕建,等.4株鸡传染性贫血病毒的分离鉴定及其遗传进化分析[J].中国预防兽医学报,2016(9):695-699.
[11]GORYO M,SUWA T,MATSUMOTO S,et al.Serial propagation and purification of chicken anaemia agent in MDCCMSB1 cell line[J].Avian Pathol,1987,16(1):149-163.
[12]YAMAGUCHI S,IMADA T,KAJI N,et al.Identification of a genetic determinant of pathogenicity in chicken anaemia virus[J].J Gen Virol,2001,82(5):1233-1238.
[13]TODD D,CONNOR T J,CALVERT V M,et al.Molecular cloning of an attenuated chicken anaemia virus isolate following repeated cell culture passage[J].Avian Pathol,1995,24(1):171-187.
[14]MEEHAN B M,TODD D,CREELAN J L,et al.Investigation of the attenuation exhibited by a molecularly cloned chicken anemia virus isolate by utilizing a chimeric virus approach[J].J Virol,1997,71(11):8362-8367.