FELINE NEUROTROPIC RETROVIRUSES

猫神经营养逆转录病毒

• 批准号: 3415042
• 负责人: LAWRENCE R WHALEN
• 金额: $ 25.64万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3415042
• 关键词: HIV infections; Retroviridae; Retroviridae disease; Schwann cells; cats; disease /disorder model; dorsal root; electrophysiology; evoked potentials; feline leukemia /sarcoma virus; genetic strain; genome; molecular cloning; natural gene amplification; nervous system infection; neural conduction; neurotropic virus; nucleic acid probes; nucleic acid sequence; spinal cord; tissue /cell culture; virus classification; virus replication

The overall goal of this investigation is to develop an animal model of the neurologic abnormalities associated with HIV infection using molecularly-clones (MC) strains of feline neurotropic retroviruses. A feline model of experimentally-reproducible, retrovirus-induced neurological disease will facilitate studies in three areas; determination of the precise cellular and electrophysiologic abnormalities directly attributable to retrovirus infection; identification of the viral genetic basis for retroviral infection of the central nervous system; and, most importantly, will allow the design and testing of therapies to counter the challenge of infections within the nervous system. Specific aims for this five year project are: first, to characterize the neurological deficits and lesions in cats experimentally inoculated with neurotropic strains of feline leukemia virus (FeLV), and to determine the cellular tropism of these FeLVs in cell culture; second, to clone neurotropic FeLV-1390, identify the molecular differences between it and the minimally pathogenic strain of retrovirus (MC-FeLV-3281) from which it arose, and then to evaluate the pathogenicity of MC-FeLV-1390; third, to use the techniques refined in the FeLV studies to identify, isolate and molecularly characterize neurotropic strains of the recently discovered feline lentivirus (FIV), followed by characterization of the pathogenicity of MC-FIVs. This model system will allow the evaluation of anti-HIV therapies. The pathogenicity of neurotropic FeLVs will be evaluated in vitro in feline spinal cord, dorsal root ganglion, and schwann cell cultures and in vivo in specific pathogen free (SPF) cats. In vitro studies will be designed to identify neurotropic strains of feline retroviruses and to determine in which cell types they replicate and are cytopathic. Initial studies will focus on further characterization of strain FeLV-1390 that we recently isolated from a neurologically abnormal cat. Evaluation of infected cats will include prospective studies of electrophysiology (sequential conduction velocities of motor and sensory nerves and of evoked spinal cord potentials), immunological testing (functional studies), sequential neurological examinations and virus isolations (cerebral spinal fluid, blood, and bone marrow), and histopathology. Molecular studies of neurotropic FeLVs will include; identification of the viral genome present in the tissues of cat 1390 and cats inoculated with virus derived from this cat; molecular cloning of the FeLV-1390 neurotropic variant; and identification of the changes in this virus related to its minimally pathogenic parental virus (MC-FeLV-3281) through nucleotide sequence analysis. Additional sequence comparisons will be made to determine if FeLV-1390 arose through acquisition of endogenous sequences. Potentially neurotropic FIVs will be identified by performing neurologic and electrophysiologic exams of naturally-occurring cases of FIV which present to the Colorado State University Veterinary Teaching Hospital. FIVs will be isolated from cats with neurologic dysfunction, and the pathogenicity of these isolates will be evaluated in vitro and in vivo in order to identify a neurotropic strain of FIV for further study. FIV will be cloned from a strain that induces neurologic dysfunction in SPF cats. A biologically defined FIV model for lentivirus-induced neurologic disease will be established using MC-FIVs, and we will begin to define the genetic structures of neurotropic FIVs.

这项调查的总体目的是开发一个动物模型 使用与HIV感染相关的神经系统异常 猫神经性逆转录病毒的分子clones（MC）菌株。 一个 实验可再生，逆转录病毒诱导的猫科动物模型 神经疾病将促进三个地区的研究； 确定精确的细胞和电生理学 直接归因于逆转录病毒感染的异常； 鉴定病毒遗传基础的逆转录病毒感染 中枢神经系统；而且，最重要的是，将允许设计和 测试疗法以应对感染挑战的疗法 神经系统。 该五年项目的具体目的是：首先要表征 猫的神经缺陷和病变实验接种了 猫白血病病毒（FELV）的神经性菌株，并确定 这些FELV在细胞培养中的细胞向向;第二，要克隆 神经性FELV-1390，确定IT和 逆转录病毒（MC-FELV-3281）的最小致病菌株从中 它出现，然后评估MC-FELV-1390的致病性；第三， 使用FELV研究中完善的技术来识别，隔离 并以分子为特征最近的神经菌株 发现的猫科动病毒（FIV），然后是表征 MC-FIV的致病性。 该模型系统将允许评估 抗HIV疗法。 将在体外评估神经性FELV的致病性 猫的脊髓，背根神经节和施旺细胞培养物以及 特定病原体（SPF）猫的体内体内。 体外研究将是 旨在鉴定猫逆转录病毒的神经性菌株和 确定它们复制哪种细胞类型并具有细胞质病。 最初的 研究将集中于菌株FELV-1390的进一步表征 我们最近从神经学异常的CAT中分离出来。 评估 感染的猫将包括对电生理学的前瞻性研究 （运动和感觉神经的顺序传导速度以及 诱发的脊髓电位），免疫测试（功能性 研究），顺序神经检查和病毒分离 （脑脊髓液，血液和骨髓）和组织病理学。 神经性FELV的分子研究将包括；识别 CAT 1390组织中存在的病毒基因组和接种的猫 源自这只猫的病毒； FELV-1390的分子克隆 神经型变体；并识别该病毒的变化 通过其最小的致病性父母病毒（MC-FELV-3281） 核苷酸序列分析。 其他序列比较将是 确定FELV-1390是否通过获得内源性而产生 序列。 通过执行神经系统，将确定潜在的神经智FIV 和自然疾病病例的电生理检查 出席科罗拉多州立大学兽医教学医院。 FIV将从具有神经功能障碍的猫中分离出来，并将 这些分离株的致病性将在体外和体内评估 为了识别FIV的神经型菌株以进行进一步研究。 FIV会 从诱导SPF猫中神经功能障碍的菌株中克隆。 慢病毒诱导的神经系统的生物定义的FIV模型 疾病将使用MC-FIV建立，我们将开始定义 神经性FIV的遗传结构。