DEVELOPING AN IN VITRO MODEL OF INFECTION OF BOVINE B CELLS WITH BLV

开发 BLV 感染牛 B 细胞的体外模型

• 批准号: 7627623
• 负责人: JEFFRY S ISAACSON
• 金额: $ 4.92万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7627623
• 关键词: Animals; Antibody Formation; B-Lymphocytes; Biological Assay; Bos taurus; Bovine Leukemia Virus; Bovine Lymphosarcoma; Bovine leukemia; Cattle; Cells; Computer Retrieval of Information on Scientific Projects Database; Diffuse Lymphoma; Flow Cytometry; Funding; Grant; Health; Human T-Cell Leukemia Viruses; In Vitro; Infection; Institution; Lymphocytosis; Methods; Numbers; Publications; RNA-Directed DNA Polymerase; Reporting; Research; Research Personnel; Resources; Retroviridae; Source; T-Lymphocyte; United States National Institutes of Health; Viral Antigens; Virus Diseases; in vitro Model; infected B cell; leukemia virus

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bovine leukemia virus (BLV) is a retrovirus that infects cattle. It is closely related to the human T cell leukemia virus. However, BLV mainly infects B cells, rather than T cells. BLV causes a B cell lymphosarcoma in about 5% of infected animals, but can also cause persistent lymphocytosis and other health problems in a larger percentage of infected cattle. In previous publications, Isaacson et al. reported that cattle experimentally infected with BLV had increased antibody responses to various non-BLV antigens, and a significantly increased number of B cells from these animals were found to express MHC-II molecules. To date, incubation of BLV-negative bovine lymphosarcoma (BL-3) cells with cell-free supernatants from BLV-infected FLK-BLV cells has resulted in the BL-3 cells becoming PCR-positive for BLV. These results are currently being confirmed, and various methods for quantitation of BLV in FLK-BLV supernatants are being investigated. Thus far, attempts to determine BLV titers by a reverse transcriptase assay, as well as by a TCID50 assay using BLV-susceptible CC81 cells, have been unsuccessful, but efforts are continuing. In addition, the BLV-infected BL-3 cells will soon be assayed by flow cytometry to determine whether levels of MHC-II expression are increased following in vitro BLV infection.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 牛白血病病毒（BLV）是一种感染牛的逆转录病毒。 它与人类T细胞白血病病毒密切相关。 然而，BLV主要感染B细胞，而不是T细胞。 BLV 会导致大约 5% 的受感染动物出现 B 细胞淋巴肉瘤，但也会导致更大比例的受感染牛出现持续性淋巴细胞增多和其他健康问题。 在之前的出版物中，艾萨克森等人。报道称，实验性感染 BLV 的牛对各种非 BLV 抗原的抗体反应增强，并且发现这些动物中表达 MHC-II 分子的 B 细胞数量显着增加。 迄今为止，将 BLV 阴性牛淋巴肉瘤 (BL-3) 细胞与来自 BLV 感染的 FLK-BLV 细胞的无细胞上清液一起孵育，可导致 BL-3 细胞成为 BLV PCR 阳性。 这些结果目前正在得到证实，并且正在研究用于定量 FLK-BLV 上清液中 BLV 的各种方法。 迄今为止，通过逆转录酶测定以及使用 BLV 敏感 CC81 细胞进行 TCID50 测定来确定 BLV 滴度的尝试尚未成功，但仍在继续努力。 此外，BLV感染的BL-3细胞很快将通过流式细胞术进行分析，以确定体外BLV感染后MHC-II表达水平是否增加。