Study on molecular mechanisms of herpesvirus entry into host cells

疱疹病毒侵入宿主细胞的分子机制研究

• 批准号: 16390138
• 负责人: INOUE Naoki
• 金额: $ 9.54万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390138/
• 关键词: herpesviruses; glycoproteins; varicella-zoster virus; cytomegalovirus; genotypes; pseudotypes; cell-cell fusion; signal responses of host cells; 感染; ヒトヘルペスウイルス8; 宿主遺伝子; 水泡性口内炎ウイルス

1. Using a cell-cell fusion assay and a VSV pseudotyping assay, it was demonstrated that herpes simplex virus type 1 (HSV-1) gB, gH, gL, gD were necessary for virus entry. Under the same condition, any combinations of glycoproteins encoded by cytomegalovirus (CMV) and varicella-zoster virus (VZV) did not yield positive results. However, any of CMV or VZV glycoproteins inhibited the HSV-1 glycoprotein mediated fusion or infection with pseudotyped VSV.2. To analyze epitomes on glycoproteins associated neutralization, a new assay using VSV pseudotyped with MLV env protein and the target glycoprotein.3. Inhibition of attachment with heparin-like molecules and inhibition of immediate early phase of infection with kinase inhibitors were evaluated by using reporter cell lines for VZV and CMV.4. A filter-based real-time PCR assay was developed for identification of CMV-positive clinical materials. CMV DNA specimens identified by the assay were used for genotyping of glycoprotein genes. There was a linkage among gH, gO, and gN genotypes, although there was no linkage of gB genotypes with those of other glycoproteins. One hypothesis is that pressure on gB by neutralization relates to the lack of linkage and alternative hypothesis is that particular combination of gH, gO, and gN are required for their functions in virus entry.5. Promoter region for activation of interferon stimulating gene 54K (ISG54K) promoter by CMV infection was delineated. The activation was dependent on CMV IE gene expression.

1.用细胞-细胞融合试验和VSV假型试验证明，单纯疱疹病毒1型（HSV-1）gB、gH、gL、gD是病毒进入所必需的。在相同条件下，巨细胞病毒（CMV）和水痘-带状疱疹病毒（VZV）编码的糖蛋白的任何组合均不产生阳性结果。然而，任何CMV或VZV糖蛋白抑制HSV-1糖蛋白介导的融合或感染假型VSV。利用MLV env蛋白和靶糖蛋白对VSV进行假型，建立了一种新的中和相关糖蛋白表位分析方法.通过使用VZV和CMV的报告细胞系来评价与肝素样分子的附着的抑制和与激酶抑制剂的感染的立即早期的抑制。建立了一种用于CMV阳性临床材料鉴定的实时荧光PCR方法。使用该检测试剂鉴定的CMV DNA标本进行糖蛋白基因的基因分型。gH、gO和gN基因型之间存在连锁关系，但gB基因型与其他糖蛋白基因型之间没有连锁关系。一种假设是中和作用对gB的压力与缺乏连接有关，另一种假设是gH、gO和gN的特定组合是它们在病毒进入中的功能所必需的。描述了CMV感染激活干扰素刺激基因54 K（ISG 54 K）启动子的启动子区。该激活依赖于CMV IE基因的表达。

项目成果

CMVの先天性感染機構

CMV先天性感染机制

• 发表时间: 2006
• 期刊: 日本臨床 64・3
• 作者: Mizutani T;Fukushi S;Saijo M;Kurane I;Morikawa S.;田口文広;Taguchi F;井上直樹;野澤直樹
• 通讯作者: 野澤直樹

Higher prevalence of human herpesvirus 8 DNA sequence and specific IgG antibodies in patients with pemphigus in China

• DOI: 10.1016/j.jaad.2004.10.882
• 发表时间: 2005-03-01
• 期刊: JOURNAL OF THE AMERICAN ACADEMY OF DERMATOLOGY
• 影响因子: 13.8
• 作者: Wang, GQ;Xu, HH;Chen, HD
• 通讯作者: Chen, HD

The impact of viral interferon regulatory factor-1 expression on responsiveness to interferon alpha and the production of infectious human herpesvirus 8 by BCBL-1 cells.

病毒干扰素调节因子 1 表达对干扰素 α 反应以及 BCBL-1 细胞产生传染性人类疱疹病毒 8 的影响。

• 发表时间: 2004
• 期刊: Journal of Virology 78
• 作者: Mizutani T;Fukushi S;Saijo M;Kurane I;Morikawa S.;田口文広;Taguchi F;井上直樹;野澤直樹;Wang GQ;N.Inoue;V.Pozharskaya
• 通讯作者: V.Pozharskaya

A real-time PCR assay using specimens on filter discs as a template for detection of cytomegalovirus in urine.

使用滤盘上的样本作为检测尿液中巨细胞病毒的模板进行实时 PCR 测定。

• 发表时间: 2007
• 期刊: J Clin Microbiol. 45
• 作者: Arguni;E.;et al.;Kumar N 他;Nozawa N 他
• 通讯作者: Nozawa N 他

A monoclonal antibody that recognizes Epstein-Barr virus nuclear antigen 2 (EBNA2) amino acids 1-58 does not react with EBNA 2 in native form, consistent with the self-association of EBNA 2 through the amino-terminus.

识别 Epstein-Barr 病毒核抗原 2 (EBNA2) 氨基酸 1-58 的单克隆抗体不会与天然形式的 EBNA 2 发生反应，这与 EBNA 2 通过氨基末端的自缔合一致。

• 发表时间: 2005
• 期刊: Arch Virol. 150・5
• 作者: G.Wang;H.Xu;Y.Wang;X.Gao;Y.Zhao;C.He;N.Inoue;H.D.Chen.;Harada S
• 通讯作者: Harada S