Development of NHP Models for Infectious Pathogens of Man

对于细胞跟踪项目，我们已经成功地开发了我们称之为连续血管内染色的方法，以确定细胞在体内24至48小时内的运输。事实证明，这项技术在确定不同亚群进出组织的相对贩运比率方面非常有用。我们计划很快将这一点与标记疫苗结合起来，以确定启动疫苗反应的细胞进入各种组织的运输率。 对于诺沃克病毒，我们已经开始从人类身上收集诺沃克病毒株的过程，我们将把它们混合在一起，对多个非人类灵长类动物进行单一挑战，试图找到一个或多个株，这些株至少在灵长类动物的肠道中显示出一些最小的复制。我们还成功地开始培养具有正确形态和细胞组成的非人类灵长类肠道动物。在VRC的合作中，我们将使用他的NV探针来识别人类和NHP中的抗原特异性B细胞，我们可以从中产生新的单抗。 最后，对于WEVEE，我们已经分离了近100种不同的非人类灵长类单抗，这三种病毒都有；使用我们的方法，我们可以选择性地识别那些对所有三种病毒都有交叉反应的抗体。我们已经证明，结合这三种病毒的抗体只会中和一种病毒--IE，这些病毒表面的中和表位是不同的。这表明，单抗治疗或预防可能需要至少两种不同的单抗来保护免受所有这三种病毒的攻击。我们与VRC的Kuong博士合作，在每个病毒尖峰的背景下结晶这些单抗，以确定病毒的完整抗原表面结构，并了解它们为什么有不同的中和表位。

For the cell tracking project, we have successfully developed the method we term serial intravascular staining to define the trafficking of cells over a 24 to 48 hour period In vivo. This technique has proven immensely useful for defining the relative trafficking rates of different subpopulations into and out of tissues. We are planning to marry this soon with labeled vaccines to define the trafficking rates of cells that initiate vaccine responses into a variety of tissues. For norovirus, we have begun the process of collecting norovirus strains from humans, and we'll mix them together in a single challenge to multiple nonhuman primates to try to find one or more strains that show at least some minimal replications in the primate gut. We've also successfully begun to grow nonhuman primate enteroids that shows the right morphology and cell composition. In collaboration with Dr Kwong at the VRC come on we will be using his NV probes to identify antigen specific B cells in both humans and in NHP, we from which we can generate novel monoclonal antibodies. Finally, for WEVEE, we have isolated almost 100 distinct nonhuman primate monoclonal antibodies two the three viruses; with our methodology, we can selectively identify those which are cross reactive to all three. We have shown that antibodies which bind all three viruses, will only neutralize one -- IE, neutralizing epitopes on the surface of these viruses are distinct. This suggests that Monoclonal antibody therapeutics or preventatives may require at least two distinct monoclonal antibodies for protection against all three viruses. In collaboration with Dr. Kwong at the VRC, we are crystalizing these monoclonal antibodies in context of each of the viral spikes in order to define the complete antigenic surface structure of the viruses and understand why they have distinct neutralizing epitopes.