BOVINE LEUKOSIS VIRUS PATHOGENESIS

本项目的长期目标是确定机制 牛白血病病毒（BLV）在牛中的致病性 人逆转录病毒相关淋巴恶性肿瘤模型。 我们 工作假设是BLV感染的T淋巴细胞是关键的 淋巴组织异常增生 这一假设将是 通过对BLV的向性进行广泛表征进行测试， 单核白细胞亚群及相关分子 导致淋巴细胞增多和/或肿瘤发展的事件。 细胞 来自所有主要淋巴区室的亚群将被 从处于疾病进展多个阶段的动物中分析 （白细胞减少、淋巴细胞增多和携带肿瘤阶段）。 单核 白细胞亚群扰动将通过流式细胞仪 细胞计数和免疫组织学。 单核细胞的病毒嗜性 亚群将通过鉴定细胞来确定 携带前病毒（Southern杂交和PCR）。 的能力 这些细胞表达BLV蛋白的能力将通过 细胞内抗原免疫荧光染色（流式细胞术） 细胞计数和免疫组织学）; 具有潜在细胞生长调节活性的BLV蛋白 （特别是反式激活蛋白，税收）将由 基于北方杂交和/或PCR的mRNA鉴定。 T和B的遗传克隆性。细胞将通过识别 IG和TCR基因重排。 BLV税收驱动T的作用 淋巴细胞产生B和/或T细胞生长因子将是 通过功能测定以及北方杂交测定- 和/或基于PCR的白细胞介素mRNA的鉴定。 协会 在获得的所有数据中，疾病进展的状态将 大大增加了我们对BLV发病机制的了解，并提供 在细胞和分子水平上， 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.