VIRAL GENOME STRUCTURE & LENTIVIRUS PATHOGENICITY

人类免疫缺陷病毒(HIV、HTLV-III、LAV、ARV)和 绵羊慢病毒(OvLV)具有共同的形态和基因组 相似之处并导致持续感染导致缓慢、 淋巴系统、呼吸道和神经系统的进行性疾病。 儿童原发性淋巴间质性肺炎(LIP) 最近，成年人与艾滋病毒感染有关。嘴唇 作为OvLV诱导的一种常见临床表现发生 可以通过以下方式在新生羔羊中迅速诱发 气管内注射OvLV。建议的目的是 研究是利用这种慢病毒感染的动物模型来 评估慢病毒基因组结构之间的关系 和致病性。为了研究人类免疫缺陷病毒的决定因素 羔羊的品系特异性致病和免疫反应， 将使用两个空斑纯化的OvLV分离株：85/34，a 能在12分钟内引起唇裂的高度裂解的分离株 周，以及84/28，一种诱导缓慢进展的分离株 体外合胞体，与绵羊肺相关 癌症和嘴唇。寄主遗传变异的影响将是 通过使用同基因同卵双胞胎羔羊来最小化 接种疫苗。七对羔羊将在出生时接种 其中一种分离物或带有介质的。将选出成对的动物 评估损伤和免疫反应的重复性 在病毒株内和株之间。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 病毒感染的自体和自体细胞的细胞毒作用 同种异体靶细胞将在5个月内进行评估。 处死后，取肺、区域淋巴结、脾、 其他器官将被检测是否存在组织学损害和 传染性病毒。此外，自发产生的干扰素 将对每只羔羊的肺淋巴细胞进行定量 与病变的发展密切相关。病毒株将是 分子克隆和核酸水平上的比较 限制性内切酶作图和杂交。病毒 将在蛋白质水平上进行分子大小的比较 主要结构蛋白的抗原性。抗原性 将使用直接约束性和竞争性进行比较 放射免疫分析、放射免疫沉淀 病毒蛋白，以及免疫印迹电泳法。 沙门氏菌核酸结构与蛋白质组成的相关性 每株病毒的致病性和免疫反应存在差异 就会被制造出来。

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.