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WANG Yinan,ZHAN Wenbin,XING Jing,JIANG Yousheng. 2008. In vivo neutralization assays by monoclonal antibodies against white spot syndrome virus in crayfish(Cambarus proclarkii). Acta Oceanologica Sinica, (2):126-132
In vivo neutralization assays by monoclonal antibodies against white spot syndrome virus in crayfish(Cambarus proclarkii)
In vivo neutralization assays by monoclonal antibodies against white spot syndrome virus in crayfish(Cambarus proclarkii)
Received:June 05, 2007  Revised:December 19, 2007
DOI:
Key words:crayfish  WSSV  monoclonal antibodies  in vivo neutralization
中文关键词:  crayfish  WSSV  monoclonal antibodies  in vivo neutralization
基金项目:The National Basic Research Program of China under contract No. 2006CB101806;the National "863" Project of China under con-tract No. 2006AA100312.
Author NameAffiliationE-mail
WANG Yinan Laboratory of Pathology and Immunology of Aquatic Animals, Laboratory of Mariculture of Ministry of Education, Ocean University of China, Qingdao 266003, China  
ZHAN Wenbin Laboratory of Pathology and Immunology of Aquatic Animals, Laboratory of Mariculture of Ministry of Education, Ocean University of China, Qingdao 266003, China wbzhan@ouc.edu.cn 
XING Jing Laboratory of Pathology and Immunology of Aquatic Animals, Laboratory of Mariculture of Ministry of Education, Ocean University of China, Qingdao 266003, China  
JIANG Yousheng Laboratory of Pathology and Immunology of Aquatic Animals, Laboratory of Mariculture of Ministry of Education, Ocean University of China, Qingdao 266003, China  
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Abstract:
      The neutralizing activities of eight monoclonal antibodies (MAbs) against white spot syndrome virus (WSSV) (2D2, 2B2, 1D2, 1D5, 1C2, 4A1, 6A4 and 6B4) were analyzed by in vivo experiments.Gills from WSSV-infected shrimp were homogenized and ten-fold serially diluted by PBS, and then incubated with MAbs (hybridoma culture supernatant), respectively.The mixture of WSSV and MAbs were injected into crayfish (Procambarus clarkii).After challenge, the death rates of crayfish were counted to determine the neutralizing activities of MAbs.At the same time, the mixture of myeloma culture supernatant and WSSV or PBS was served as positive or negative control, respectively.The results showed that, at each virus dilution, the mean time to death of the crayfish injected with MAb-treated virus was significantly longer than that in the positive control, though they all showed 100% mortality within 25 d, and meanwhile, few crayfish died in the negative control.Among the eight MAbs, 2D2, 2B2, 1D2 and 1D5, especially the former two, delayed the mortality significantly, and 1C2, 4A1 and 6A4 delayed the mortality as well but not so efficiently, while MAb 6B4 was efficient only when the virus concentration increased.The results indicated that the anti-WSSV MAbs can neutralize WSSV in different virus dilutions.
中文摘要:
      The neutralizing activities of eight monoclonal antibodies (MAbs) against white spot syndrome virus (WSSV) (2D2, 2B2, 1D2, 1D5, 1C2, 4A1, 6A4 and 6B4) were analyzed by in vivo experiments.Gills from WSSV-infected shrimp were homogenized and ten-fold serially diluted by PBS, and then incubated with MAbs (hybridoma culture supernatant), respectively.The mixture of WSSV and MAbs were injected into crayfish (Procambarus clarkii).After challenge, the death rates of crayfish were counted to determine the neutralizing activities of MAbs.At the same time, the mixture of myeloma culture supernatant and WSSV or PBS was served as positive or negative control, respectively.The results showed that, at each virus dilution, the mean time to death of the crayfish injected with MAb-treated virus was significantly longer than that in the positive control, though they all showed 100% mortality within 25 d, and meanwhile, few crayfish died in the negative control.Among the eight MAbs, 2D2, 2B2, 1D2 and 1D5, especially the former two, delayed the mortality significantly, and 1C2, 4A1 and 6A4 delayed the mortality as well but not so efficiently, while MAb 6B4 was efficient only when the virus concentration increased.The results indicated that the anti-WSSV MAbs can neutralize WSSV in different virus dilutions.
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