EVALUATION OF ONCOGENIC FACTORS RELEVANT TO DEVELOPING SAFE HERPESVIRUS VACCINES

与开发安全疱疹病毒疫苗相关的致癌因素的评估

• 批准号: 3770332
• 负责人: A RAZZAQUE
• 金额: --
• 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/3770332
• 关键词: AIDS; Herpesviridae vaccine; athymic mouse; bone marrow transplantation; carcinogen testing; carcinoma; chronic fatigue syndrome; cytomegalovirus; human herpesvirus 6; human tissue; lymphoma; microorganism immunology; molecular cloning; neoplasm /cancer genetics; nucleic acid sequence; oncogenes; oncoproteins; protein sequence; regulatory gene; transfection /expression vector; tumor antigens; viral carcinogenesis; virus DNA; virus genetics; virus protein

Genomes and subgenomic clones of herpesviruses, human cytomegalovirus (HCMV and human herpesvirus-6 (HHV-6), were studied for oncogenic potential in order to evaluate their role in human cancers. CMV: Multiple regions of CMV strain Towne were studied for transforming functions by introducing DNA into rodent cells and human cells, and assayin the selected cells for tumorigenicity by growth in agarose or in nude mice. In this study, the major immediate early gene (IE1), which is a candidate subunit vaccine was also included. We identified two domains, mtrII and mtrIII, in the CMV genome that could transform cells to tumorigenicity. Th mtrII transformed human cells could produce poorly differentiated carcinoma in mice. Both domains contain several protein coding sequences. Data from the digestion of these domains with restriction enzymes suggested that a 79 amino acid (aa) protein in mtrII and a 270 aa protein in mtrIII might be responsible for tumorigenicity. Further mapping is in progress. We found that the IE1 gene is not involved in transformation. HHV-6: Similar studies were applied to HHV-6 to elucidate its role in huma lymphomas especially in AIDS and bone marrow transplant patients. Genomic DNA of HHV-6 isolates from AIDS patients (GS and Z29) and from chronic fatigue syndrome patients (GD) could transform mouse fibroblasts. Two DNA clones (21 kb and 8.7 kb) of GS isolate could transform human keratinocytes HHV-6 DNA, specific to 8.7 kb region, has been detected in an immunoblastic lymphoma by hybridization. Further studies are in progress to determine th specific genes involved in transformation. Identification and characterization of herpesvirus oncogenes are important to safety evaluatio and to development of CMV and other herpesvirus vaccines as well as to development of herpesvirus-based vectors for gene therapy. Since IE1 of CM is found non-tumorigenic in our assay, it may be safely used as a candidate subunit vaccine. If a tumor antigen is identified, a specific diagnostic reagent can be developed for CMV or HHV-6 related malignant diseases.

疱疹病毒、人巨细胞病毒（HCMV）的基因组和亚基因组克隆 和人类疱疹病毒-6（HHV-6）的致癌潜力进行了研究， 以评估它们在人类癌症中的作用。 CMV：研究CMV株Towne的多个区域用于转化 通过将DNA导入啮齿动物细胞和人类细胞， 通过在琼脂糖中或在裸鼠中生长，选择细胞的致瘤性。 在这项研究中，主要立即早期基因（IE 1），这是一个候选基因， 还包括亚单位疫苗。 我们确定了两个域，mtrII和 mtrIII，在CMV基因组中，可以将细胞转化为致瘤性。 日 mtrII转化的人细胞可产生低分化癌 对小鼠 两个结构域都含有几个蛋白质编码序列。 数据从 用限制性内切酶消化这些结构域表明， mtrII中的氨基酸（aa）蛋白和mtrIII中的270 aa蛋白可能是 负责致瘤性。 进一步的测绘工作正在进行。我们发现 IE 1基因不参与转化。 HHV-6：对HHV-6进行了类似的研究，以阐明其在人类免疫中的作用。 淋巴瘤，特别是艾滋病和骨髓移植患者。 基因组 来自AIDS患者（GS和Z29）和来自慢性HIV感染者的HHV-6分离物的DNA 疲劳综合征患者（GD）可转化小鼠成纤维细胞。 两个DNA GS分离物的21 kb和8.7kb克隆可转化人角质形成细胞 HHV-6DNA特异于8.7kb区域，已在一个免疫母细胞中检测到， 淋巴瘤杂交。 进一步的研究正在进行中，以确定 参与转化的特定基因。 识别和 疱疹病毒癌基因的特征对安全性评价很重要 以及CMV和其它疱疹病毒疫苗的开发， 开发用于基因治疗的基于疱疹病毒的载体。 自CM的IE 1 在我们的检测中发现无致瘤性，它可以安全地用作候选物 亚单位疫苗 如果确定了肿瘤抗原， 可开发用于CMV或HHV-6相关恶性疾病的试剂。