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Preventive measures of Mark's desease (MD) vaccine break caused by very virulent MD virus.

由强毒力MD病毒引起的马克氏病（MD）疫苗失效的预防措施。

基本信息

批准号：
01860041
负责人：
MIKAMI Takeshi
金额：
$ 11.33万
依托单位：
The university of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

项目摘要

For prevention of Marek's disease (MD) caused by Marek's disease virus (MDV), monovalent vaccine of herpesvirus of turkey has been widely used for chickens in the field. Recently, however, MD caused by very virulent MDV (vvMDV) was observed in vaccinated chicknes. The final aim of the present study is to develop MD vaccine such as recombiant, polyvalent or component vaccine for controlling the outbreak of MD due to vvMDV. The following results were obtained.1) Arecombinant baculovirus expressing the A antigen (rBV-A) of MDV was constructed. In Spodoptera frugiperda (Sf) cells infectedwith the rBV-A, A antigen (A Ag) expression was localized to the cell surface. By immunoprecipitation of ^<35>S labelled A Ag with a monoclonal antibody to A Ag, a large amount of the A Ag(58-68kDa) was dtected of the rBV-A infected cells but a small amount of the antigen (46-54kDa) was found in the culture supenatant of infected cells. The reconminant A Ag was shown to be reactive with sera of MDV-infected chickens. In sera of chickens immunized with the A Ag, MDV-specific antibody reactive with MDV-infected cells was found.2) A gene encoding a homolog of glycoprotein B of herpes simplex virus (gB homolog) has been identified on the MDV genome. The recombinant baculovirus encoding the gB homolog (rBV-B) of MDV expressed the protein in the cytoplasm and on the surface of Sf cells infected with the rBV-B, when examined by immunofluorescent assay using anti-B sera from chickens or rabbits, and monoclonal antibody against B antigen. The expressed protein had three molecular sized (88/100, 64 and 54kDa).3) Chicks thymectomized and injected with either monoclonal antibody to chicken CD4 of CD8 were infected with vvMDV. The CD8-depleed chcks developed both nerve enlargement and visceral tumor, whereas the CD4-depleted chicks showed only nerve enlargement.

为了预防由马立克病病毒（MDV）引起的马立克病（MD），火鸡疱疹病毒单价疫苗已被广泛应用于鸡群。然而，最近在接种疫苗的鸡中观察到由非常毒力的MDV （vvMDV）引起的MD。本研究的最终目的是开发重组疫苗、多价疫苗或组分疫苗，以控制由vvMDV引起的MD暴发。得到了以下结果：1)构建表达MDV A抗原的重组杆状病毒（rBV-A）。在被rBV-A感染的夜蛾（Spodoptera frugiperda, Sf）细胞中，A抗原（A Ag）在细胞表面表达。用A Ag单克隆抗体免疫沉淀标记A Ag的^<35>S，在rBV-A感染细胞中检测到大量的A Ag(58-68kDa)，而在感染细胞的培养上料中检测到少量的A Ag（46-54kDa）。残留的aag与感染mdv的鸡血清有反应。在经A抗原免疫的鸡血清中，发现了与mdv感染细胞有反应的mdv特异性抗体。2)在MDV基因组上发现了一个编码单纯疱疹病毒糖蛋白B同源基因（gB同源基因）。重组杆状病毒编码MDV的gB同源物（rBV-B），在感染了rBV-B的Sf细胞的细胞质和表面表达该蛋白，用鸡或兔的抗B血清和抗B抗原的单克隆抗体进行免疫荧光检测。表达的蛋白有3个分子量（88/ 100,64和54kDa）。3)经胸腺切除并注射鸡CD8 CD4单克隆抗体的雏鸡感染vvMDV。cd8缺失的雏鸡出现神经增大和内脏肿瘤，而cd4缺失的雏鸡只出现神经增大。