Animal and Laboratory Core

动物和实验室核心

基本信息

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
项目类别：
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

项目摘要

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 病毒储备对于本申请中提出的所有实验，感染它们。核心B将生成BLT（骨髓肝- 胸腺）人源化小鼠，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艾滋病毒人类免疫反应。 BLT小鼠是通过手术植入免疫缺陷小鼠肾囊下的人胎儿胸腺和肝脏组织，同时自体人类造血干细胞的静脉移植。该协议允许人类 T 细胞在这些小鼠中成熟的细胞是通过自体人类胸腺组织而不是异种小鼠胸腺。这种自体胸腺教育似乎可以大大改善 T 细胞功能，我们和其他人在 BLT 小鼠中观察到这一点，从而大大改善了人类 B 细胞功能。核心 B 已经具备向计划调查人员提供重组良好数量的能力他们打算在 HIVRAD 应用中研究 BLT 小鼠。 Core B 还将对携带 HIV 的 BLT 小鼠进行疫苗接种和/或感染，以及后续的工作从这些接种疫苗和/或感染艾滋病毒的小鼠身上采集血液和组织。核心B人员有与合作研究人员在 BLT 小鼠中进行 HIV 研究的丰富经验。安全性强已制定计划，以尽量减少进行 HIV 感染和分析的核心 B 人员的暴露风险感染 HIV 的 BLT 小鼠，并已制定应急程序，以防发生任何暴露情况。核心B 还将通过开发低剂量重复粘膜来改善 HIV 感染的 BLT 模型挑战模型。重复的挑战将使小鼠能够更好地感染艾滋病毒剂量代表人类异性接触中遇到的那些。核心 B 还将为所有计划研究人员提供病毒学支持。核心 B 将生成并滴定用于感染 BLT 小鼠的 HIV 库存，并通过 qRT-PCR 测量 HIV 感染小鼠的血浆病毒载量，使用完善的 Core B 协议。核心 B 已证明有能力提供 HIVRAD 应用实验需要多种 B、A 和 C 分支 HIV 毒株。因此，核心 B 能够很好地支持所有动物和病毒学要求 HIVRAD 计划中提议的调查。