VIRAL GENOME STRUCTURE & LENTIVIRUS PATHOGENICITY

人类免疫缺陷病毒（艾滋病毒、HTLV-III、LAV、ARV）和 绵羊慢病毒（OvLV）具有相同的形态和基因组 相似之处，并导致持续感染，导致缓慢， 淋巴、呼吸和神经系统的进行性疾病。 儿童原发性淋巴间质性肺炎（LIP）， 成人最近与艾滋病毒感染有关。 唇 作为OvLV诱导的常见临床表现发生 疾病的绵羊，并可以迅速诱发新生羔羊， 血管内注射OvLV。 建议的目的 研究是使用这种慢病毒感染的动物模型， 评估慢病毒基因组结构之间的关系 和致病性。 为了研究 在羔羊中的菌株特异性致病和免疫应答， 将使用两种空斑纯化的OvLV分离物：85/34，a 能够在12小时内诱导严重LIP高度裂解的田间分离物 周，84/28，一种诱导缓慢进行性 体外合胞体，并与绵羊肺 癌和LIP。 宿主遗传变异的影响将是 通过使用同基因单卵双胞胎羔羊， 接种。 七对羔羊将在出生时接种 一种分离物或培养基。 将选出一对动物 为了评估病变和免疫反应的可重复性， 在病毒株内和病毒株之间。 血清抗体水平， 支气管肺泡细胞的变化，病毒血症的发展，以及 病毒感染的自体和 同种异体靶细胞将在5个月的时间内进行评价。 处死后，取肺、区域淋巴结、脾， 将检测其他器官是否存在组织学病变， 传染性病毒 此外，自发的干扰素产生 将对每只羔羊的肺淋巴细胞进行定量， 与病变发展相关。 病毒株将会 在核酸水平上进行分子克隆和比较， 限制性内切酶作图和杂交。 的病毒 将在蛋白质水平的分子大小方面进行比较 和主要结构蛋白的抗原特征。 抗原 比较将使用直接绑定和竞争 放射免疫测定，放射性标记的放射免疫沉淀 病毒蛋白质和免疫印迹法。 核酸结构与蛋白质组成的相关性 各菌株具有致病性和免疫反应差异 将被制作。

Human immunodeficiency virus (HIV, HTLV-III, LAV, ARV) and ovine lentiviruses (OvLV) share morphological and genomic similarities and cause a persistent infection resulting in slow, progressive disease of lymphoid, respiratory, and nervous systems. Primary lymphoid interstitial pneumonia (LIP) in children and adults has recently been associated with HIV infection. LIP occurs as a common clinical manifestation of OvLV-induced disease in sheep and can be rapidly induced in neonatal lambs by injecting OvLV intratracheally. The purpose of the proposed research is to use this animal model of lentivirus infection to evaluate the relationship between lentivirus genomic structure and pathogenicity. To investigate the viral determinants of strain specific pathogenic and immunologic responses in lambs, two plaque purified OvLV isolates will be employed: 85/34, a highly lytic field isolate capable of inducing sever LIP within 12 weeks, and 84/28, an isolate that induces slowly progressive syncytia in vitro and is associated with ovine pulmonary carcinoma and LIP. The effects of host genetic variation will be minimized by use of isogeneic monozygotic twin lambs for inoculation. Seven pairs of lambs will be inoculated at birth with one of the isolates or with media. Pairs of animals will be chosen to evaluate reproducibility of lesions and immune responses both within a viral strain and between strains. Serum antibody levels, changes in bronchoalveolar cells, development of viremia, and cell-mediated cytotoxicity of virus-infected autologous and allogeneic target cells will be evaluated over a 5 month period. After sacrifice, lung, regional lymph nodes, spleen, and selected other organs will be assayed for presence of histologic lesions and infectious virus. Additionally, spontaneous interferon production by pulmonary lymphocytes will be quantitated for each lamb and correlated with lesions development. The viral strains will be molecularly cloned and compared at the nucleic acid level by restriction endonuclease mapping and hybridization. The viruses will be compared at the protein level in terms of molecular size and antigenic features of major structural proteins. Antigenic comparisons will be made using direct binding and competition radioimmunoassay, radioimmunoprecipitation of radiolabelled viral proteins, and immunoblotting of electrophoresed proteins. Correlations of nucleic acid structure and protein composition of each strain with pathogenicity and immune response differences will then be made.