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| 題 名 | Passive Immunity to White Spot Syndrome Virus (WSSV) in Penaeus Monodon Treated with Monoclonal Antibodies of the Heterologously Produced VP28 Viral Envelope Protein |
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| 作 者 | Chen, Jyh-yih; Anbarasu, Kumarasamy; Chen, Chin-yu; Lee, Ying-chuan; Nan, Fan-hua; Kuo, Ching-ming; | 書刊名 | Journal of Marine Science and Technology |
| 卷 期 | 18:1 2010.02[民99.02] |
| 頁 次 | 頁145-152 |
| 分類號 | 387.13 |
| 關鍵詞 | White spot syndrome virus; Penaeus monodon; Monoclonal antibody; Immersion; Oral administration; |
| 語 文 | 英文(English) |
| 英文摘要 | ABSTRACT White spot syndrome virus (WSSV) is a highly pathogenic and prevalent virus affecting shrimp culture worldwide including Taiwan. In the present study, the viral envelope protein, VP28, gene of the WSSV was cloned into a pET28a expression vector. VP28 was expressed as a protein with a 6-histidine tag in Escherichia coli and purified by a Ni-NTA column. Antiserum was raised against this recombinant VP28 protein in BALB/C mice, and it recognized the VP28 protein in purified virions and recombinant proteins. The antiserum was mixed with arginine (100 ppm), lysine (168 ppm), and phenylalanine (88 ppm), and the titer of the antibody was 20,000-fold/ml. A series of monoclonal antibodies was tested for their ability to neutralize WSSV infectivity by immersion and oral administration. Our results proved that the antibody developed against the VP28 protein could efficiently prevent or control WSSV infection in post-larval and juvenile stages by immersion and oral administration. Furthermore, the postlarvae obtained from the antibody-treated group had > 60% survival after 20 days of challenge, illustrating that the antibody produced from heterologously expressed VP28 may specifically provide passive immunity against the WSSV pathogen. Therapeutic studies illustrated that shrimp losses couldspecific memory may exist in the defense system of this invertebrate species, which reacted more efficiently after encountering an antigenic parasite [7]. Extracted plasma from surviving shrimp after a WSSV infection can be used to neutralize WSSV from 20 to 60 days after infection [18]. These results suggest that an adaptive immune response may exist in shrimp, and protection against WSSV can possibly be induced in shrimp by a vaccination. Furthermore, a strategy was developed and adopted for protecting P. monodon against WSSV by oral vaccination [14]. Recently, a few studies have made efforts to develop polyclonal antibodies (pAbs) against specific viral peptides such as VP28 and VP19 and have obtained promising results with indoor experiments [17, 8]. Nevertheless, considering all these previous reports, in the present study, a monoclonal antibody (mAb) was developed against the VP28 peptide alone in high titers using hybridoma technology with a mouse model, and its prophylactic and treatment efficiencies were evaluated via immersion and oral routes in P. monodon, a highly susceptible species to WSSV. |
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