BIOLOGICAL AND PHYSICAL PROPERTIES OF FRIEND VIRUS

朋友病毒的生物学和物理特性

• 批准号: 3179009
• 负责人: ROBERT J ECKNER
• 金额: $ 18.99万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-30 至 1988-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3179009
• 关键词: B lymphocyte; Friend virus; bone marrow transplantation; cell growth regulation; clone cells; density gradient ultracentrifugation; erythroleukemia; hematopoietic stem cells; host neoplasm interaction; immunofluorescence technique; latent virus infection; leukopoiesis; neoplasm /cancer immunology; oncogenes; radiation leukemia; viral leukemogenesis; virus assembly; virus cytopathogenic effect; virus genetics; virus replication

The overall objective of this project is to determine if defective Friend spleen focus-forming virus (SFFV) carries genetic information which is able to function as a hemopoietic self-renewal gene when introduced into the pluripotent hemoietic stem cell (CFU-S) of murine bone marrow cells. Major Specific Aims; SFFV constructions. To construct a transmissible retrovirus consisting of the molecularly cloned Friend SFFV and an altered mouse dihydrofolate reductase (dhfr) sequence. The presence of a single mutant dhfr sequence serves as a dominant-acting selectable gene. Persistence if engineered defective SFFV. Suspensions of murine bone marrow cells enriched for CFU-S will be exposed to infectious SFFV-dhfr to determine if the host range of this engineered, defective virus includes hemopoietic cells. Virus-treated marrow will be injected into lethally-irradaited histocompatible adult recipients. These mice will be monitored for latent SFFV persistence. SFFV and stem cells self-renewal. Using donor marrow which are predominantly SFFV+ CFU-S, to probe for SFFV-induced alterations in host hematopoiesis and stem cell renewal. SFFV+ marrow cells will be retrieved from latently infected mice and serially transplanted to determine their ability to self-renew and protect or rescue lethally-irradiated recipients from total hemopoietic failure and death. To insure repopulation by predominantly SFFV+ CFU-S, we shall treat reconstituted mice with a drug regimen if methotrexate (Mtx). Identify of an SFFV self-renewal gene. To determine if the envelope (env) sequences which encode gp52sfv are required for the induction of extended CFU-S self renewal. A number of well-defined SFFV insertion-deletion mutants will be reconstituted with dhfr, pseudotypes with helper, and used to infect CFU-S. Latent persistence will be confirmed using an SFFV representative probe, while the SFFV+ stem cells will be monitored for self-renewal capacity via serial transplantation into lethally irradiated syngeneic adult mice. Vector transfer of self-renewal may eventually be exploited to the benefit of the host whether he/she be suffering from an aquired or congenital defect in hemopoiesis erytghromyeloid and lymphoid.

该项目的总体目标是确定是否存在缺陷 Friend 脾病灶形成病毒（SFFV）携带遗传信息，能够 当导入到造血细胞中时，可作为造血自我更新基因发挥作用 小鼠骨髓细胞的多能造血干细胞（CFU-S）。 主要具体目标； SFFV 结构。 构建可传播的逆转录病毒，其组成为 分子克隆的 Friend SFFV 和改良的小鼠二氢叶酸 还原酶 (dhfr) 序列。 单个突变体 dhfr 序列的存在 作为显性作用的选择基因。 如果设计有缺陷的 SFFV，则持久性。 鼠骨悬液 富含 CFU-S 的骨髓细胞将暴露于传染性 SFFV-dhfr 确定该工程缺陷病毒的宿主范围是否包括 造血细胞。 经过病毒处理的骨髓将被注射到 经致死照射的组织相容性成年受者。 这些老鼠将 监测潜在 SFFV 持续性。 SFFV 和干细胞自我更新。 使用供体骨髓 主要是 SFFV+ CFU-S，用于探测 SFFV 诱导的宿主变化 造血和干细胞更新。 将回收 SFFV+ 骨髓细胞 来自潜伏感染的小鼠并连续移植以确定它们的 自我更新和保护或拯救受到致命辐射的接受者的能力 完全造血衰竭和死亡。 确保人口重新增加 主要是 SFFV+ CFU-S，我们将用药物治疗重组小鼠 甲氨蝶呤 (Mtx) 治疗方案。 SFFV 自我更新基因的鉴定。 判断是否有信封（env） 编码 gp52sfv 的序列是诱导延伸所必需的 CFU-S自我更新。 一些明确定义的 SFFV 插入删除 突变体将用 dhfr 重建，假型用 helper 重建，并使用 感染CFU-S。 将使用 SFFV 确认潜在持久性 代表性探针，同时将监测 SFFV+ 干细胞 通过连续移植到致命辐射中的自我更新能力 同基因成年小鼠。 自我更新的载体转移最终可能会被利用以带来好处 宿主是否患有后天性或先天性的 红骨髓和淋巴造血功能缺陷。