Animal and Laboratory Core

动物和实验室核心

• 批准号: 8492624
• 负责人: TODD M ALLEN
• 金额: $ 79.1万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-07 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8492624
• 关键词: Address; Adjuvant; Adopted; Animal Model; Animals; Antigens; Autologous; B-Lymphocytes; Biological Models; Blood; Blood specimen; Bone Marrow; CD34 gene; Cell physiology; Cells; Control Groups; Dose; Education; Emergency Situation; Ensure; Event; Exposure to; Fibrous capsule of kidney; Generations; HIV; HIV Infections; Harvest; Hematopoietic; Hematopoietic stem cells; Heterosexuals; Human; Human Resources; Immune; Immune response; Immunodeficient Mouse; Implant; Infection; Instruction; Intervention; Intravenous; Investigation; Laboratory Animals; Liver; Mature T-Lymphocyte; Measures; Methods; Modeling; Monitor; Mus; Names; Operative Surgical Procedures; Osteomyelitis; Plasma; Positioning Attribute; Procedures; Protocols documentation; Research Personnel; Risk; Safety; Sampling; Scientist; Stem cells; T-Lymphocyte; Thymic Tissue; Thymus Gland; Tissue Sample; Tissues; Transplantation; Vaccinated; Vaccination; Vaccines; Vagina; Viral; Viral Load result; Virus; Work; bone; design; experience; fetal; implantation; improved; in vivo; mouse model; nanoparticle; novel; prevent; programs; reconstitution; research study; transmission process; vaginal transmission; vector

Core B, the Animal and Laboratory Core, will provide both the humanized mice, and the HIV viral stocks to infect them with, for all experiments proposed in this application. Core B will generate BLT (bone marrowliver- thymus) humanized mice, which provide one of the recently improved humanized mouse models of HIV infection. BLT mice can be infected with HIV by vaginal transmission, and are capable of producing functional anti-HIV human immune responses. BLT mice are generated by the surgical implantation of human fetal thymic and liver tissue under the renal capsules of immunodeficient mice, concurrently with the intravenous transfer of autologous human hematopoietic stem cells. This protocol allows the human T cells that mature in these mice to be educated by autologous human thymic tissue rather than by the xenogenic mouse thymus. This autologous thymic education appears to result in the much improved T cell function, and consequently much improved human B cell function, that we and others have observed in BLT mice. Core B has already well-established capacity to provide Program investigators with the numbers of wellreconstituted BLT mice that they propose to study in this HIVRAD application. Core B will also perform the vaccinations and/or infections of BLT mice with HIV, and the subsequent harvesting of blood and tissues from these vaccinated and/or HIV-infected mice. Core B personnel have extensive experience in performing HIV studies in BLT mice with collaborating investigators. A strong safety plan is in place to minimize the risk of exposure to Core B personnel performing HIV infections and analyses of HIV-infected BLT mice, and emergency procedures are in place in case any exposure does occur. Core B will also work to improve the BLT model of HIV infection, by developing a low-dose repeated mucosal challenge model. Repetitive challenges will enable the mice to be infected with HIV doses that better represent those encountered in human heterosexual exposure. Core B will also provide virological support to all Program investigators. Core B will generate and titer the stocks of HIV used to infect BLT mice, and measure plasma viral loads in HIV-infected mice by qRT-PCR, using well-established Core B protocols. Core B has in place the demonstrated capacity to provide the multiple Clade B, A and C HIV strains that the experiments of this HIVRAD application will require. Core B thus is well-positioned to support all of the animal and virological requirements of the investigations proposed in this HIVRAD program.

核心B，即动物和实验室核心，将提供人源化小鼠和HIV病毒储备 来感染它们，用于本申请中提出的所有实验。核心B将产生BLT（骨髓肝- 胸腺）人源化小鼠，其提供了最近改进的HIV人源化小鼠模型之一 感染BLT小鼠可以通过阴道传播感染HIV，并且能够产生 功能性抗HIV人类免疫应答。BLT小鼠通过手术植入 免疫缺陷小鼠肾包膜下的人胎胸腺和肝组织，同时 自体造血干细胞的静脉内移植。该方案允许人类T细胞 在这些小鼠中成熟，通过自体人类胸腺组织而不是异种胸腺组织进行训练， 小鼠胸腺这种自体胸腺训练似乎导致T细胞功能大大改善， 因此大大改善了人类B细胞的功能，这是我们和其他人在BLT小鼠中观察到的。 核心B已经具备了向项目调查人员提供大量经过良好重组的 BLT小鼠，他们建议在这个HIVRAD应用研究。 核心B还将进行BLT小鼠的疫苗接种和/或HIV感染，以及随后的免疫接种。 从这些接种疫苗的和/或HIV感染的小鼠收获血液和组织。核心B人员有 与合作研究者在BLT小鼠中进行HIV研究的丰富经验。强有力的安全 已制定计划，以最大限度地降低执行HIV感染和分析的核心B人员的暴露风险 感染艾滋病毒的BLT小鼠，并采取紧急程序，以防任何接触发生。核心B 还将致力于改善艾滋病毒感染的BLT模型，通过开发低剂量的重复粘膜 挑战模式重复的挑战将使小鼠感染艾滋病毒的剂量更好， 代表人类异性接触中遇到的那些。 核心B还将为所有项目研究者提供病毒学支持。核心B将产生并滴度 用于感染BLT小鼠的HIV原液，并通过qRT-PCR测量HIV感染小鼠中的血浆病毒载量， 使用完善的核心B协议。核心B已证明有能力提供 该HIVRAD应用的实验将需要的多个进化枝B、A和C HIV毒株。 因此，核心B处于良好的位置，以支持本发明的所有动物和病毒学要求。 在这个艾滋病防治计划中提出的调查。