EXPRESSION OF ROTAVIRUS OUTER CAPSID PROTEIN VP4 BY A BACULOVIRUS RECOMBINANT

杆状病毒重组体表达轮状病毒外衣壳蛋白 VP4

• 批准号: 3790876
• 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/3790876
• 关键词: Baculoviridae; Reoviridae disease; Rotavirus; alleles; antiserum; antiviral antibody; capsid; diarrhea; gene expression; genetic manipulation; genetic polymorphism; genetic strain; infection; rotavirus vaccines; serotyping; tissue /cell culture; transfection /expression vector; virus antigen; virus classification; virus genetics; virus protein

Rotaviruses are ubiquitous agents that cause diarrheal disease in the young of not only humans but also many animal species. Several candidate vaccines against human rotavirus (HRV) disease are currently being evaluated with special emphasis on the VP7 outer capsid protein, which is a major protective antigen. Currently, eight different serotypes have been established among HRVs based on the antigenic specificity of VP7. The outer capsid protein, VP4, is also a major protective antigen, but relatively little is known about the antigenic relationships and extent of polymorphism of the VP4 of various HRVs. In previous studies, it was shown by sequence analysis that at least four alleles of the VP4 gene are present among HRVs. One allele of VP4 is present on strains recovered from patients with symptomatic infection. These strains had been classified as VP7 serotype 1 (strains KU and Wa), VP7 serotype 3 (strain P), or VP7 serotype 4 (strain VA70). These rotavirus strains, designated as VP4 genetic group 1, possess a highly conserved VP4 protein that exhibits 93-97% amino acid identity. A second allele of VP4 is present in HRVs belonging to VP7 serotype 2(strains DS-1 and RV-5) the exhibit 98% VP4 amino acid identity. These strains, designated as VP4 genetic group 2, were also recovered from symptomatic infections. Moreover, the VP4s of genetic group 2 are most closely related to the corresponding proteins of group 1, exhibiting an 89- 90% amino acid homology. A third allele of VP4 is present in HRV strains that were recovered from asymptomatic neonatal infections and that belong to VP7 serotype 1 (M37), VP7 serotype 2 (1076), VP7 serotype 3 (McN), or VP7 serotype 4 (ST3). the VP4s of these strains (designated VP4 genetic group 3) are highly related, as indicated by 95-97% amino acid identity. The VP4s of the third genetic group exhibit only a 75% amino acid identity with the corresponding VP4 proteins of groups 1 and 2. A fourth allele is present on the HRV K8 strain (belonging to VP7 serotype 1) that appears to have a unique VP4 gene exhibiting only 64-65% amino acid identity with the VP4 proteins of viruses of VP4 genetic group 1, 2, or 3. In a recent study, four different VP4 alleles present on HRV strains KU (genetic group 1), DS-1 (genetic group 2), 1076 (genetic group 3), and K8 (genetic group 4) were separately expressed in Spodoptera frugiperda (Sf9) cells using the baculovirus vector Autographa californica nuclear polyhedrosis virus. Antisera produced in response to immunization with these recombinant VP4 proteins allowed us to define the extent and distribution of VP4 antigenic polymorphism among rotaviruses currently circulating in humans.

轮状病毒是一种普遍存在的病原体，可引起年轻人的腹泻 不仅是人类，还有许多动物物种。 若干候选 针对人类轮状病毒（HRV）疾病的疫苗目前正在 特别强调了VP7外壳蛋白的评价， 主要的保护性抗原 目前，已经有八种不同的血清型被 基于VP7的抗原特异性在HRV中建立。 的 外壳蛋白VP4也是一种主要的保护性抗原， 关于抗原关系和免疫反应的程度知之甚少。 各种HRV的VP4多态性。 在以前的研究中， 通过序列分析，VP4基因的至少四个等位基因是 存在于HRV中。 VP4的一个等位基因存在于从患有 症状感染。 这些菌株被归类为VP7血清型 1型（菌株KU和Wa）、VP7血清型3型（菌株P）或VP7血清型4型（菌株 VA70）。 这些轮状病毒株，命名为VP4基因组1，具有 一种高度保守的VP4蛋白，具有93 - 97%的氨基酸同一性。 VP4的第二个等位基因存在于属于VP7血清型的HRV中 2株（DS-1和RV-5）的VP4氨基酸序列具有98%的同源性。 这些 命名为VP4基因组2的菌株也从 有症状的感染 此外，遗传组2的VP4最多， 与组1的相应蛋白质密切相关，表现出89- 90%的氨基酸同源性。 VP4的第三个等位基因存在于HRV毒株中 从无症状新生儿感染中恢复， VP7血清型1（M37）、VP7血清型2（1076）、VP7血清型3（McN），或 VP7血清型4（ST3）。 这些菌株的VP4（命名为VP4基因 组3）高度相关，如95 - 97%氨基酸同一性所示。 第三遗传群的VP4仅显示75%的氨基酸同一性 与组1和组2的相应的VP4蛋白一起。 第四个等位基因是 存在于HRV K8毒株（属于VP7血清型1）上， 具有独特的VP4基因，其仅显示出64 - 65%的氨基酸与 VP4基因组1、2或3的病毒的VP4蛋白。 在最近的一项研究中，HRV毒株KU上存在四种不同的VP4等位基因， （遗传群1）、DS-1（遗传群2）、1076（遗传群3）和K8 （遗传群4）分别在草地贪夜蛾（Spodoptera frugiperda，Sf9）中表达， 使用杆状病毒载体Autographa californica核 多角体病毒 免疫后产生的抗血清 这些重组VP4蛋白使我们能够确定其程度， 目前轮状病毒VP4抗原多态性的分布 在人体内循环。