BOVINE LEUKOSIS VIRUS PATHOGENESIS

这个项目的长期目标是明确机制(S) 牛白血病病毒(BLV)对牛的致病作用 人类逆转录病毒相关淋巴系统恶性肿瘤的模型。我们的 工作假设是BLV感染的T淋巴细胞是关键 与异常淋巴组织增殖有关。这一假设将是 通过对BLV的趋向性进行广泛的特征测试 单核白细胞亚群及其相关分子 导致淋巴细胞增多和/或肿瘤发展的事件。细胞 所有主要淋巴细胞室的亚群将是 从疾病发展的多个阶段的动物身上进行分析 (白血期、淋巴细胞期和荷瘤期)。单核 白细胞亚群的扰动将通过流动来识别 细胞学和免疫组织学检测。单核细胞的病毒嗜性 亚群将通过细胞鉴定来确定 携带前病毒(Southern杂交和聚合酶链式反应)。的能力 这样的细胞表达BLV蛋白将由 细胞内抗原免疫荧光染色(Flow 细胞学和免疫组织学)；细胞表达的证据 具有潜在细胞生长调节活性的BLV蛋白 (具体地说，反式激活蛋白，Tax)将通过 基于Northern杂交和/或聚合酶链式反应的mRNA鉴定。 T和B细胞的遗传克隆性将通过鉴定 免疫球蛋白和TCR基因重排。BLV税收驱动型T的作用 淋巴细胞产生B和/或T细胞生长因子(S) 通过功能分析和Northern杂交确定- 和/或基于聚合酶链式反应的白介素mRNA鉴定。联谊会 在所有获得的数据中，随着疾病的进展， 极大地提高了我们对BLV发病机制的了解，并为 在细胞和分子水平上，对 逆转录病毒介导的淋巴细胞增殖失调。

The long term goal of this project is to define the mechanism(s) of bovine leukosis virus (BLV) pathogenesis in cattle as an animal model for human retrovirus-associated lymphoid malignancies. Our working hypothesis is that BLV-infected T lymphocytes are pivotal to the abnormal lymphoproliferation. This hypothesis will be tested by extensive characterization of the tropism of BLV for mononuclear leukocyte subpopulations and associated molecular events that lead to lymphocytosis and/or tumor development. Cell subpopulations from all major lymphoid compartments will be analyzed from animals at multiple stages of disease progression (aleukemic, lymphocytotic & tumor-bearing stages). Mononuclear leukocyte subpopulation perturbations will be identified by flow cytometry and immunohistology. Viral tropism for mononuclear subpopulations will be determined by identification of cells harboring provirus (Southern hybridization & PCR). The ability of such cells to express BLV proteins will be determined by immunofluorescence staining of intracellular antigen (flow cytometry & immunohistology); evidence of cellular expression of BLV proteins with potential cell-growth regulatory activity (specifically the trans-activating protein, tax) will be by Northern hybridization- and/or PCR-based identification of mRNA. The genetic clonality of T and B. cells will be by identification of Ig and TCR gene rearrangement. A role for BLV tax-driven T lymphocyte production of B and/or T cell growth factor(s) will be determined by functional assays as well as Northern hybridization- and/or PCR-based identification of interleukin mRNA. Association of all data obtained, with the state of disease progression, will greatly increase our understanding of BLV pathogenesis and provide additional insight, at the cellular and molecular level, into retrovirus-mediated dysregulation of lymphocyte proliferation.