Animal and Laboratory Core

核心B，动物和实验室核心，将提供人源化小鼠和艾滋病毒病毒库存 以感染他们，对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，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艾滋病毒，并能够产生 功能性抗HIV人类免疫反应。BLT小鼠是由外科植入的 免疫缺陷小鼠肾包膜下的人胎胸腺和肝组织，同时 静脉移植自体人造血干细胞。这项协议允许人类T细胞 在这些小鼠中成熟的是由自体的人类胸腺组织而不是由异种的 小鼠胸腺。这种自体胸腺教育似乎大大改善了T细胞功能， 因此，我们和其他人在BLT小鼠身上观察到的人类B细胞功能得到了极大的改善。 核心B已经有能力向计划调查人员提供经过良好重组的数量 他们计划在HIVRAD应用程序中研究BLT小鼠。 CORE B还将为感染艾滋病毒的BLT小鼠进行疫苗接种和/或感染，以及随后的 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级人员有 与合作研究人员合作，在BLT小鼠身上进行艾滋病毒研究的丰富经验。强大的安全性 是否制定了计划，以最大限度地减少核心B人员感染艾滋病毒和进行分析的风险 对于感染艾滋病毒的BLT小鼠，已经制定了应急程序，以防发生任何接触。核心B 还将致力于改进艾滋病毒感染的BLT模型，通过开发一种低剂量重复粘膜 挑战模型。重复的挑战将使小鼠能够感染更好的艾滋病毒剂量 代表那些在人类异性接触中遇到的。 核心B还将为所有项目调查人员提供病毒学支持。核心B将生成并滴定 用来感染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， 使用成熟的核心B协议。核心B已具备证明的能力，可提供 该HIVRAD应用程序的实验将需要多个分支B、A和C HIV毒株。 因此，核心B处于有利地位，以支持所有动物和病毒学的要求 在这项HIVRAD计划中提出的调查。

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.