IMPACT OF ART ON DC & TREG RESPONSES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本研究将探讨SIV感染期间口腔免疫功能的完整性，以及长期ART如何影响口腔免疫功能，（白色念珠菌或TLR 3配体polyIC）将用于在ART期间测量口腔免疫完整性。为了跟踪这些动物的粘膜和全身免疫应答，除了已经购买的新动物外，我们一直在使用来自其他研究的回收动物的样品优化流式细胞术和功能测定。这涉及建立流式细胞术测定以记录外周血和淋巴组织中的T细胞和DC亚群及其活化状态。已经建立了功能测定法，其中将粘膜组织片（相对于血细胞）与刺激物（念珠菌属或polyIC）一起培养以监测细胞因子/趋化因子应答。将通过Luminex/ELISA和CC/CK mRNA的PCR测量此类应答。这些化验大多已到位，基线取样已于1月开始。计划在3月用SIVmac 239攻击动物的扁桃体，ART将在2周后开始。治疗组包括：未治疗的未感染动物、ART治疗的未感染动物、感染的未治疗动物和感染的ART治疗动物。将随时间监测血液和粘膜样本，以测量病毒和免疫参数。6-7 SIV感染后3个月，通过用白色念珠菌芽管或polyIC处理动物并测量局部与全身免疫和病毒载量来评估口腔免疫功能。大约8周后，将停止ART。这将有助于我们了解口腔免疫系统在感染期间如何变化，以及长期ART是否会影响这种生物学。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This study will explore the integrity of oral immune function during SIV infection and how this is impacted by long term ART, where application of potential stimuli (Candida albicans or the TLR 3 ligand polyIC) will be used to measure oral immune integrity during ART. In order to follow the mucosal and systemic immune responses of these animals, we have been optimizing the flow cytometry and functional assays using samples from recycled animals from other studies, in addition to the new animals that have already been purchased for this work. This has involved establishing the multicolor flow assays to document T cell and DC subsets and their activation states in peripheral blood and lymphoid tissues. Functional assays have been established where pieces of mucosal tissues (vs blood cells) are cultured with stimuli (Candida or polyIC) to monitor the cytokine/chemokine responses. Such responses will be measured by luminex/ELISA and PCR for CC/CK mRNA. Most of these assays are in place and the baseline sampling commenced in January. The animals are scheduled to be challenged with SIVmac239 across the tonsils in March and ART will start 2 weeks later. The treatment groups include: untreated uninfected animals, ART-treated uninfected animals, infected untreated animals, and infected ART-treated animals. Blood and mucosal samples will be monitored over time to measure viral and immune parameters. 6-7 months after SIV infection the oral immune function will be assessed by treating the animals with either Candida albicans germ tube or polyIC and measuring local vs systemic immunity and viral loads. Approximately 8 weeks later ART will be stopped. This will help us understand how the oral immune system changes during infection and whether long term ART impacts this biology.