DEVELOPMENT OF RECOMBINANT BCG-VP8 VACCINES

轮状病毒是引起严重腹泻病的主要病原体 世界各地的婴幼儿。中和抗体 已证明轮状病毒是由两种外源病毒独立诱导的 衣壳蛋白、VP7和VP4；这些抗体与 对疾病的抵抗能力。 利用杆状病毒表达系统成功地表达了 不同轮状病毒株的VP4。在这个系统中合成的蛋白质 根据其与多克隆和 现有的单抗(MAb)。此外，对豚鼠进行了免疫 用这种蛋白质开发的抗体可以中和病毒的传染性 有效免疫轮状病毒。此外，对豚鼠的免疫 表达的VP4诱导抗体的VP8或VP5裂解亚单位 这也中和了轮状病毒的传染性。通过交叉试验获得的结果- 免疫沉淀和中和试验表明这些抗血清 VP4的VP8亚单位包含主要的抗原位点 通过VP4中和轮状病毒。 这项研究的一个主要目的是鉴定沙门氏菌VP4蛋白 小鼠轮状病毒EB株(VP7血清型3型)及其粘膜免疫的确定 在同源小鼠模型中对这种外衣壳蛋白的反应。我们 选择了人类结核病疫苗卡介苗作为粘膜载体- 盖林(卡介苗)，一种牛分枝杆菌的减毒菌株，已被 广泛且安全地在人类身上使用。卡介苗接种通常是在 出生后，只需一剂即可产生持久的免疫反应。它是 也是一种高效的佐剂。

Rotaviruses are major etiologic agents of severe diarrheal disease in infants and young children throughout the world. Neutralizing antibodies to rotaviruses have been shown to be induced independently by two outer capsid proteins, VP7 and VP4; these antibodies are associated with resistance to illness. The baculovirus expression system has been used to successfully express the VP4 of various rotavirus strains. The protein synthesized in this system exhibits native conformation based on its reactivity with polyclonal and available monoclonal antibodies (MAbs). In addition, guinea pigs immunized with such proteins developed antibodies that neutralized infectivity of the immunizing rotavirus efficiently. Moreover, immunization of guinea pigs with the expressed VP8 or VP5 cleavage subunit of VP4 induced antibodies that also neutralized rotavirus infectivity. Results obtained by cross- immunoprecipitation and by neutralization assay with these antisera suggest that the VP8 subunit of VP4 contains the major antigenic sites responsible for the neutralization of rotavirus via VP4. A major objective of this study was to characterize the VP4 protein of the murine rotavirus Eb strain (VP7 serotype 3 and define the mucosal immune response to this outer capsid protein in the homologous mouse model. We chose as a mucosal vector the human tuberculosis vaccine, bacille Calmette- Guerin (BCG), an attenuated strain of mycobacterium bovis that has been used extensively and safely in humans. BCG vaccine is commonly given at birth and elicits a long-lasting immune response with a single dose. It is also a highly effective adjuvant.