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Mechanism of latent infection of human herpesvirus 6

人类疱疹病毒6型潜伏感染机制

基本信息

批准号：
07457077
负责人：
YAMANISHI Koichi
金额：
$ 4.8万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457077/
关键词：
Latency HHV-6 Immediate-early gene Reactivation IE1 IE2

项目摘要

We have been attempting to analyze the latency of human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7) in vitro and in vivo.1) In vivo, study ; the latency and reactivation of HHV-6 and HHV-7 were surveyed in throat of children of different age-groups and adults by polymerase chain reaction. The detection rate of HHV-6 DNA was the highest in children aged 1-year-old and decreased with age, while the detection rate of HHV-7 DNA increased with age and reached a maximum in adults. HHV-6B was detected in almost all samples. When the antibody prevalence was determined in the 4 groups of children. HHV-6 antibody was detected in almost all children aged 6-12 months after birth, and children were infected with HHV-7 later than HHV-6.2) In vitro study ; in order to identify the regulatory genes of HHV-6 the whole DNA of HHV-6B was sequenced and one of immediate early genes was analyzed. The immediate-early protein IE 1 ; 958 amino acids of HHV-6B strain HST were expressed as b-galactosidase fusion proteins in E coli. Using Western blot analysis, an antigenic region of the IE 1 protein was mapped between residues 340 and 505. Monospecific antibody raised against the fusion protein reacted with apparent molecular weights of 155 and 170kDa, and was detected as granular fluorescence in nuclei of infected cells by IFA.3)By using the latency system in vitro, we have identified a latency specific transcript, which is found in monocyte/macrophage infected with HHV-6B.This mRNA was specifically transcribed from the region of immediate-early (IE) gene and found in the peripheral blood in normal children, suggesting that a mRNA is transcribed in monocyte/macrophage even in vivo.

本研究对人疱疹病毒6型（HHV-6）和人疱疹病毒7型（HHV-7）的潜伏期进行了体外和体内研究。1）体内研究：采用聚合酶链反应（PCR）技术，检测了不同年龄组儿童和成人咽喉部HHV-6和HHV-7的潜伏期和再激活情况。HHV-6 DNA检出率在1岁儿童中最高，随年龄增长而下降; HHV-7 DNA检出率随年龄增长而上升，在成人中达到最高。HHV-6 B在几乎所有的样品中被检测到。在4组儿童中测定抗体流行率。6-12月龄儿童几乎全部检出HHV-6抗体，儿童感染HHV-7晚于HHV-6。2）体外研究：为了鉴定HHV-6的调控基因，对HHV-6 B全基因进行测序，并对其中一个即刻早期基因进行分析。HHV-6 B株HST的即早蛋白IE 1（958个氨基酸）在大肠杆菌中表达为β-半乳糖苷酶融合蛋白。使用Western印迹分析，IE 1蛋白的抗原区域定位在残基340和505之间。针对融合蛋白的单特异性抗体的表观分子量为155和170 kDa，并通过IFA在感染细胞的细胞核中检测到颗粒状荧光。3）通过体外潜伏系统，我们鉴定了一种潜伏特异性转录物，HHV-6 B感染的单核/巨噬细胞中发现该mRNA，该mRNA特异性地转录自HHV-6 B感染的单核/巨噬细胞的立即早期（IE）区域，该基因在正常儿童的外周血中发现，这表明即使在体内，mRNA也在单核细胞/巨噬细胞中转录。