Development and Application of Fish Iridovirus Vaccine

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Project title

Code/計畫編號

96農科-1.1.2-漁-F1(2)

Translated Name/計畫中文名

魚類虹彩病毒疫苗之研發與應用

Project Coordinator/計畫主持人

Hsin-Yiu Chou

Funding Organization/主管機關

Council of Agriculture,Executive Yuan

Co-Investigator(s)/共同執行人

張繼堯

Department/Unit

Department of Aquaculture

Website

https://www.grb.gov.tw/search/planDetail?id=1484574

Year

2007

Start date/計畫起

01-07-2007

Expected Completion/計畫迄

31-12-2007

Bugetid/研究經費

760千元

ResearchField/研究領域

漁業

疫苗開發是解決魚類病毒性疾病的重要防治對策之一，因此本計劃擬針對石斑虹彩病毒研發多醣類佐劑口服疫苗和次單位疫苗。首先測定多醣類對提升石斑魚非特異性免疫的效果，再選擇適當濃度和不活化疫苗混合製成口服製劑。對石斑魚進行餵食免疫後，以不同魚類虹彩病毒進行攻毒試驗，評估此口服疫苗的保護效果。另外進行次單位疫苗的開發，藉由石斑魚虹彩病毒在兔、羊、雞與石斑魚體內的免疫反應為依據，利用免疫接種後所產生的抗虹彩病毒多株抗體，進行其對虹彩病毒抗原性之分析；並選擇可能的石斑魚虹彩病毒開放讀架基因來製備重組蛋白以作為候選次單位疫苗。目前已完成虹彩病毒GIV010R (63 kDa)，GIV042L (62 kDa)等次單位疫苗蛋白質在大腸桿菌/pET表現系統的表現與純化，另外病毒的主要鞘蛋白也順利以四個分段完成重組蛋白MCP-1 (16 kDa)，MCP-2 (13 kDa)，MCP-3 (13 kDa)及MCP-4 (11 kDa)的表現與純化。這些虹彩病毒候選次單位疫苗以石斑魚苗進行初步效價評估，發現其具有40至60%的相對保護效價，而魚苗血清抗體專一性之分析，則發現MCP-2，MCP-3，MCP-4和GIV042L具有血清抗體專一性，因此，將利用所製備的各類多株抗體來分析石斑魚虹彩病毒主要鞘蛋白的抗原決定區。另外，也將以不同次單位疫苗組成、不同疫苗劑量、不同接種次數、以及不同攻毒劑量等方法，利用石斑魚吋苗來進行次單位疫苗效價及保護力的田間試驗評估分析。 Vaccine development is one of the major approaches of controlling viral diseases in aquaculture. Therefore, in this project, aiming at practical uses, attempts were made to develop glucan adjuvanted oral vaccine and subunit vaccine against grouper iridovirus simultaneously. Firstly, the effectiveness of glucan on enhancing nonspecific immune response in grouper was determined. Then the optimal concentration of glucan will be supplemented with TGIV inactivated vaccine to manufacture glucan adjuvanted oral TGIV vaccine. This oral vaccine delivery system will use to immunize grouper, and efficacy against different fish iridoviruses will be determined in vivo by vaccination-challenge protection test. Further, we plan to develop subunit vaccine for grouper iridovirus. We will prepare polyclonal antibody from rabbit, goat, chicken and grouper for grouper iridovirus, and analyze their antigenicity. Also, we still prepare the candidate for the possible open reading frame of grouper iridovirus. Until date, GIV010R (63 kDa) and GIV042L (62 kDa) had been expressed in E. coli-pET system and purified from Ni-NTA column. The four polypeptide of GIV MCP (major capsid protein), including MCP-1 (16 kDa), MCP-2 (13 kDa), MCP-3 (13 kDa) and MCP-4 (11 kDa), were successfully expressed and purified in same system. After assessing the titer of the subunit vaccines on grouper fry, the relative protection could reach 40% to 60%. MCP-2, MCP-3, MCP-4 and GIV042L showed higher antigenicity of grouper iridovirus against vaccines immunized grouper serum. Therefore, we will analyze the epitope of grouper GIV MCP with developed polyclonal antibodies. Another, we will understand the titer of subunit vaccine and its potential protection in grouper fry with different subunit vaccine component, dosage, inoculated frequency and virus challenge in field experiments.

Keyword(s)

虹彩病毒
口服不活化疫苗
主要鞘蛋白
外套蛋白
次單位疫苗
石斑魚
Iridovirus
Oral Inactivated Vaccine
Major Capsid Protein
Envelope Protein
Subunit Vaccine
Grouper

DSpace CRIS

Development and Application of Fish Iridovirus Vaccine

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