EXPRESSION OF ROTAVIRUS OUTER CAPSID PROTEIN VP4 BY RECOMBINANT ADENOVIRUS

重组腺病毒表达轮状病毒外衣壳蛋白 VP4

• 批准号: 3803273
• 负责人: M GORZIGLIA
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3803273
• 关键词: Adenoviridae; Rotavirus; gel electrophoresis; immunoprecipitation; laboratory rat; molecular cloning; oral administration; protein sequence; tissue /cell culture; virus envelope; virus protein

The outer capsid protein VP4 of porcine rotavirus OSU was chosen as a model to study the immunogenicity of this protein expressed by an adenovirus vector. The ability of adenovirus to be administered orally makes the potential use of adenovirus vectors for vaccine production particularly attractive. In addition, live adenovirus vaccines have already been shown to be safe in humans when administered orally in enteric coated capsules. The full-length cDNA of the VP4 of the porcine rotavirus OSU strain was cloned into adenovirus type 5 at a location downstream of the E3 promoter. In this viable recombinant the rotavirus VP4 sequence was substituted for sequences in the adenovirus E3 region. The plaque- purified recombinant (Ad5-OSU VP4), expressed an antigenically authentic VP4 protein as determined by immunoprecipitation. The 84 kDa protein had the same electrophoretic mobility as the native VP4 of OSU rotavirus grown in MA104 cells. Preliminary results from a study in which cotton rats were infected intranasally with live Ad5-OSU VP4 indicated that the animals developed a low level of neutralizing antibodies to the OSU porcine rotavirus.

选择猪轮状病毒OSU的外衣壳蛋白VP4作为 研究该蛋白表达的免疫原性的模型 腺病毒载体。 腺病毒口服给药的能力 使腺病毒载体有可能用于疫苗生产 特别有吸引力。 此外，腺病毒活疫苗还具有 已被证明口服给药时对人类是安全的 肠溶胶囊。 猪轮状病毒OSU株VP4的全长cDNA为 克隆到 E3 下游位置的 5 型腺病毒中 发起人。 在这个可行的重组体中，轮状病毒VP4序列是 取代腺病毒E3区中的序列。牌匾—— 纯化的重组体（Ad5-OSU VP4），表达抗原真实性 通过免疫沉淀法测定VP4蛋白。 84 kDa 蛋白质 与 OSU 轮状病毒的天然 VP4 具有相同的电泳迁移率 在 MA104 细胞中生长。 一项研究的初步结果 大鼠鼻内感染活 Ad5-OSU VP4 表明 动物产生了低水平的 OSU 中和抗体 猪轮状病毒。