Analysis of antigenic properties of human herpesvirus 7 and the host immune responses

人疱疹病毒7型抗原特性及宿主免疫反应分析

• 批准号: 10670285
• 负责人: YAMADA Masao
• 金额: $ 1.54万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670285/
• 关键词: human herpesvirus; dot-blot-neutralization assay; seroepidemiology; transferred antibody; seroconversion; monoclonal antibody; phosphoprotein; リン酸化蛋白; ドットブロット中和抗体測定法; HHV-6,-7の血清疫学

To elucidate antigenic properties of human herpesviruses 6 and 7 (HHV-6 and -7), three monoclonal antibodies (Mabs) against HHV-6 and 13Mabs against HHV-7 were established and characterized by radio-immunoprecipitation. The 40-kDa phosphoprotein by recognized by a Mab (TK17) was the product of HHV-7 U27 gene. Among these Mabs, a Mab(IK3) which recognizes the 135-kDa late polypeptide of HHV-6 and several Mabs (TK3 and others) which recognize the 125-kDa late polypeptide of HHV-7 were selected to monitor virus growth by a dot blot antigen-detection method. Using the dot blot method, we established a neutralizing antibody assay for these viruses. The dot-blot neutralization assay is easy to perform, is highly reproducible and objective when compared with the conventional methods based on cytopathology or immunofluorescence for determining neutralization endpoints. Using this method, 55 sera from healthy adults were examined. In most individuals, neutralizing antibody titers against HHV-7 were much higher than those against HHV-6. Elevated neutralizing titers against HHV-7 might be attributed to continuous booster effects by persistent HHV-7 production in saliva. Furthermore, we monitored neutralizing antibody titers against these viruses in 15 children with documented history of exanthem subitum. Transferred antibody titers against these viruses from mothers were readily demonstrated by the neutralization test. Transferred antibody titers against HHV-7 were higher and remained longer after birth than those of HHV-6, and these findings were in accord with clinical observation that HHV-6 infection usually occurs earlier than HHV-7 infection. It is notable that no cross-reactive elevation of heterologous neutralizing antibody titer was observed.

为阐明人类疱疹病毒6型和7型（HHV-6和HHV-7）的抗原特性，建立了3株抗HHV-6单克隆抗体和13株抗HHV-7单克隆抗体，并采用放射免疫沉淀法进行了鉴定。单克隆抗体TK 17识别的40 kDa磷蛋白是HHV-7 U27基因的产物。在这些单克隆抗体中，选择识别HHV-6的135-kDa晚期多肽的单克隆抗体（IK 3）和识别HHV-7的125-kDa晚期多肽的几种单克隆抗体（TK 3和其它），通过斑点印迹抗原检测方法监测病毒生长。用斑点杂交法建立了中和抗体检测方法。与基于细胞病理学或免疫荧光的常规中和终点测定方法相比，斑点印迹中和试验易于执行，重现性高，客观性好。用此方法检测了55份健康成人血清。在大多数个体中，针对HHV-7的中和抗体滴度远高于针对HHV-6的中和抗体滴度。针对HHV-7的中和滴度升高可能归因于唾液中持续产生HHV-7的持续增强效应。此外，我们还监测了15名有皮疹病史的儿童的中和抗体滴度。通过中和试验很容易证明来自母亲的抗这些病毒的转移抗体滴度。HHV-7的抗体滴度高于HHV-6，且在出生后抗体滴度维持时间较长，这与临床观察HHV-6感染通常早于HHV-7感染的现象雅阁。值得注意的是，未观察到异源中和抗体滴度的交叉反应性升高。

项目成果

山田雅夫: "第8章 臓器感染症:皮膚粘膜系,泌尿生殖系,神経系 コンパクト微生物学,小熊惠二,東匡伸編"南江堂、東京(分担部分:PP148-152,172-177,178-183). 209 (1999)

Masao Yamada：“第 8 章器官传染病：粘膜皮肤系统、泌尿生殖系统、神经系统紧凑微生物学，由 Keiji Oguma 和 Masanobu Higashi 编辑”Nankodo，东京（部分贡献：PP148-152、172-177、178-183）。 (1999)

Tsukazaki T. and 5 authors: "Development of a dot-blot neutralizing assay for HHV-6 and -7 using specific monoclonal antibodies"J. Virol. Methods.. 73. 141-149 (1998)

Tsukazaki T. 和 5 位作者：“使用特异性单克隆抗体开发 HHV-6 和 -7 的斑点印迹中和测定法”J.

Yamada, M: "Comparative biology of human herpesviruses 6 and 7. (in Japanese)"Uirusu. 48. 39-44 (1998)

Yamada, M：“人类疱疹病毒 6 和 7 的比较生物学。（日语）”Uirusu。

Yamada, M.: "Herpesvirus infection --- primary infection, latent infection and recrudescence --- (in Japanese)"Rinshyo to Kenkyu. 76. 6-10 (1999)

Yamada, M.：“疱疹病毒感染——原发感染、潜伏感染和复发——（日语）”Rinshyo to Kenkyu。

山田雅夫: "ウイルスの潜伏感染と活性化 1.単純ヘルペスウイルスとその患者"治療学. 32. 597-601 (1998)

Masao Yamada：“病毒潜伏感染和激活1.单纯疱疹病毒及其患者”治疗学32。597-601（1998）。