Studies on the T-cell epitopes of herpesviruses

疱疹病毒T细胞表位的研究

基本信息

批准号：
08456144
负责人：
OKAZAKI Katsunori
金额：
$ 4.8万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

An escape mutant of bovine herpesvirus 1 (BHV1) was obtained in the presence of monoclonal antibody against glycoprotein B (gB) , which required complement for virus neutralization. The mutant was also resistant to other 3 monoclonal antibodies against gB,which exhibited complement-independent neutralizing activity. Amino acid substitution of gB of the mutant is now being analyzed. The cytotoxic activity of peripheral blood lymphocytes from BHV1-infected cattle is also being investigated using the mutant-infected primary culture as a target to study whether the amino acid substitution affects recognition by T cells.To develope more effective and safer vaccine against infectious bovine rhinotracheitis, intranasal immunization of cattle with subunit vaccines containing baculovirus-expressed gC and/or gD of BHV1 was performed. Cholera toxin B subunit was added to the vaccines as amucosal adjuvant. Cattle immunized intranasally with the vaccines produced neutralizing antibodies against BHV1 in the nasal secresions and scarecely shed the virus after challenge with 105.0PFU of BHV1. When cattle were challenged with 107.8PFU of the virus, they showed less severity of clinical signs and less virus shedding.Mice intranasally administrated with plasmid expressing gD of Aujeszky's disease virus (ADV) produced antibodies against the virus in the trachea-lung washes and resisted to intranasal challenge with a lethal dose of the virus. Mice intranasally administrated with gC-expressing plasmid also resisted to the virus challenge. Since no antiody was detected in the nasal secresions, trachea-lung washes or serum of the mice, gC expressed in the respiratory mucosal cells may induce cellular immunity to protect from the challenge.

在抗糖蛋白 B (gB) 的单克隆抗体存在下获得了牛疱疹病毒 1 (BHV1) 的逃逸突变体，该抗体需要补体来中和病毒。该突变体还对其他3种抗gB单克隆抗体具有抗性，表现出补体非依赖性中和活性。目前正在分析突变体gB的氨基酸取代。还以突变体感染的原代培养物为靶点，研究BHV1感染牛的外周血淋巴细胞的细胞毒活性，以研究氨基酸取代是否影响T细胞的识别。开发更有效、更安全的针对传染性牛鼻气管炎的疫苗，用含有杆状病毒表达的BHV1的gC和/或gD的亚单位疫苗对牛进行鼻内免疫。霍乱毒素 B 亚基作为粘膜佐剂添加到疫苗中。用疫苗进行鼻内免疫的牛在鼻分泌物中产生了针对 BHV1 的中和抗体，并且在用 105.0 PFU 的 BHV1 攻击后几乎没有排出病毒。当牛受到 107.8 PFU 病毒攻击时，它们表现出较轻的临床症状和较少的病毒脱落。小鼠鼻内给予表达 Aujeszky 病病毒 (ADV) gD 的质粒，在气管肺洗液中产生针对该病毒的抗体并产生抵抗用致死剂量的病毒进行鼻内攻击。鼻内给予gC表达质粒的小鼠也能抵抗病毒的攻击。由于在小鼠的鼻分泌物、气管肺洗液或血清中未检测到抗体，因此呼吸道粘膜细胞中表达的gC可能诱导细胞免疫以免受攻击。