ENHANCED B VIRUS DIAGNOSIS/CONTROL IN PRIMATE COLONIES

加强灵长类动物群体中的 B 病毒诊断/控制

• 批准号: 3421472
• 负责人: Julia K Hilliard
• 金额: $ 22.09万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-15 至 1992-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3421472
• 关键词: Betaherpesvirinae; Macaca; Primates; animal colony; communicable disease control; communicable disease diagnosis; gene expression; humoral immunity; immunoglobulin G; immunoglobulin M; immunologic assay /test; immunologic techniques; immunological substance; in situ hybridization; molecular cloning; monoclonal antibody; serology /serodiagnosis; simian virus; tissue /cell culture; virus DNA; virus antigen; virus diseases; virus protein

The objective of this five year renewal program is to develop enhanced serological and virological test systems for B virus (Herpesvirus simiae) as well as other simian herpesviruses, to unambiguously identify B virus infection in humans, macaques, or other exposed nonhuman primates. We will identify structural and functional properties of B virus, and closely related herpesviruses to develop a multifaceted diagnostic system. We propose development of the following assay systems: (1) a rapid enzyme-immunoassay for serological identification of B virus specific antibodies using recombinant B virus components to replace currently required B virus propagation, (2) a rapid antigen detection system to identify the presence of replication competent B virus present in human and nonhuman primate tissues (lesional or nonlesional sites), and (3) molecular analysis using in situ hybridization techniques to identify primates with potential for reactivation of latent B virus. To accomplish the development of these systems we propose the following specific aims; (1) identify the polypeptides/glycoproteins of B virus to which human (n=6) and macaque (n=50) virus hosts direct a specific humoral response (igM and igG), (2) identify these B virus polypeptides(s)/glycoprotein(s) which contain the greatest abundance of B virus specific sequences or epitopes, (3) purify these B virus polypeptides(s)/glycoproteins(s) and subsequently, use the selected epitopes to enhance monoclonal antibody production, (4) identify the polypeptide/glycoprotein specificity of produced polyclonal monospecific antibodies and monoclonal antibodies and use these antibody reagents for purification of selected, specific, immunoreactive epitopes, (5) identify B virus specific loned DNA fragments from our B virus genomic library, (6) express these fragments in a high efficiency Autographa californica vector system, (7) develop rapid assay(s) systems using these recombinant expression products, (8) use the specific DNA fragments to identify optimum in situ probes for localization of B virus in both infected cells in culture as well as from directly sampled tissues, and (9) develop serological and virological assay systems utilizing the selected molecularly characterized B virus components from aims 1-8 for development of rapid and analytical B virus diagnostics in humans and macaques (or other exposed nonhuman primate species), including trial of these reagents to: a) identify risk factors for B virus infection and detectable shedding events within a defined breeding group of rhesus monkeys located at the California Primate Research Center and b) correlate the presence of B virus antibody, detectable B virus shedding, and latent B virus infections within trigeminal and lumbosacral dorsal root ganglia of rhesus monkeys from this population. The results of the research proposed in this renewal application will provide the basis for implementing the safest possible operating standards for NIH and other federally supported animal resource colonies and their personnel. Adoption of these standards will enhance management, prevention and control of B virus infections in humans and other primate species. Development of these diagnostic systems is especially important in preventing further zoonotic disease in humans now that macaque usage is increasing in response to the applicability of this species in AIDS research.

这个五年续签计划的目的是发展增强 B病毒的血清学和病毒学测试系统（疱疹病毒 simiae）以及其他猿猴疱疹病毒，明确地识别 B人类，猕猴或其他暴露的非人类的B病毒感染 灵长类动物。 我们将确定B的结构和功能特性 病毒和密切相关的疱疹病毒以开发多方面 诊断系统。 我们建议开发以下测定法 系统：（1）用于血清学鉴定的快速酶 - 免疫测定 使用重组B病毒成分的B病毒特异性抗体的 替换当前所需的B病毒传播，（2）快速抗原 检测系统以确定复制能力b的存在 人类和非人类灵长类组织中存在的病毒（病变或 非静态位点）和（3）分子分析使用原位 杂交技术以识别具有潜力的灵长类动物 潜在B病毒的重新激活。 为了完成这些发展 我们提出以下特定目标的系统； （1）确定 B病毒（n = 6）和猕猴的B病毒的多肽/糖蛋白 （n = 50）病毒托有直接的特定体液反应（IgM和IgG），（2） 识别这些B病毒多肽（S）/糖蛋白（S） B病毒特异性序列或表位的最大丰度，（3）纯化 这些B病毒多肽（S）/糖蛋白（S），然后使用 选定的表位以增强单克隆抗体的产生，（4） 确定产生的多肽/糖蛋白特异性 多克隆单特异性抗体和单克隆抗体并使用 这些抗体试剂用于纯化所选的特定， 免疫反应性表位，（5）识别B病毒特异性DNA 来自B病毒基因组文库的片段（6）表达这些片段 在高效率的加州矢量系统中，（7）开发 使用这些重组表达产物的快速测定系统，（8） 使用特定的DNA片段来识别最佳原位探针 B病毒在培养和 从直接采样的组织中，（9）发展血清学和 利用选定分子的病毒学测定系统 AIMS 1-8的B病毒成分表征了快速发展 和人类和猕猴的分析B病毒诊断（或其他 暴露于非人类灵长类动物），包括对这些试剂的试验： a）确定B病毒感染和可检测的脱落的危险因素 在位于 加利福尼亚灵长类动物研究中心和b）将b的存在相关联 病毒抗体，可检测的B病毒脱落和潜在B病毒 三叉神经和腰椎背根神经节中的感染 来自这个人群的恒河猴。 研究结果 在此续签申请中提出的建议将为 为NIH和其他其他实施最安全的运营标准 联邦支持动物资源殖民地及其人员。 采用这些标准将增强管理，预防和 控制人类和其他灵长类动物中B病毒感染。 这些诊断系统的开发在 现在，猕猴的使用是 响应该物种在艾滋病中的适用性而增加 研究。