DNA VACCINE FOR INDUCTION OF MUCOSAL IMMUNITY

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们假设，随着粘膜趋化因子与DNA质粒免疫的包含，将诱导更强的粘膜免疫应答，并且这种应答将对应于针对SIV攻击的增加的功效。 实验1：20只雌性印度恒河猴（Macaca mulatta）预先用DNA和各种趋化因子免疫5次。 在2010年，所有20只免疫动物加上6只未免疫动物用SIVsmE 660阴道内攻击，每周两次，持续两周。 在攻毒后的特定时间采集血样和粘膜灌洗液（包括十二指肠、肺和阴道的洗液），并运送至宾夕法尼亚大学进行免疫应答分析。 攻毒后还采集了血样和粘膜活检（包括十二指肠和阴道），并在TNPRC进行分析，以监测病毒感染和免疫应答。 攻击的结果如下：6只未免疫的对照动物中的5只、仅用DNA处理的5只动物中的3只、用DNA+cTack处理的5只动物中的4只、用DNA+Teck处理的5只动物中的2只和用DNA+Meck处理的5只动物中的4只被感染。 然而，与未免疫对照组相比，DNA+Meck动物的血浆病毒载量峰值大大降低。 所有感染的动物（n=18）在2010年被安乐死;所有未感染的动物（n=8）将用于下一个实验。 实验二：根据实验1中攻毒结果获得的数据，本实验中使用了16只新恒河猴和来自实验1的8只未感染恒河猴。 动物将接种5次SIV DNA + Teck、Flu DNA +/- Meck或C.Diff DNA +/- Meck。 最后一次接种后，所有动物将用SIVsmE 660攻毒。 将采集血液和粘膜样本，以监测病毒感染和免疫应答。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We hypothesize that, with the inclusion of mucosal chemokines with the DNA plasmid immunizations, a stronger mucosal immune response will be induced and that this response will correspond to increased efficacy against SIV challenge. Experiment 1: Twenty female Indian origin Rhesus macaques (Macaca mulatta) were previously immunized five times with DNA and various chemokines. In 2010, all 20 immunized animals plus 6 unimmunized animals were intravaginally challenged with SIVsmE660 twice a week for two weeks. Blood samples and mucosal lavages (including washes of the duodenum, lung and vagina) were collected at specified times after challenge and shipped to the University of Pennsylvania for analysis of immune responses. Blood samples and mucosal biopsies (including duodenum and vagina) were also collected after challenge and analyzed at the TNPRC to monitor viral infection and immune responses. Results of the challenge are as follows: 5 of 6 unimmunized control animals, 3 of 5 animals treated with DNA only, 4 of 5 animals treated with DNA+cTack, 2 of 5 animals treated with DNA+Teck and 4 of 5 animals treated with DNA+Meck became infected. However, the peak plasma virus loads of the DNA+Meck animals were greatly reduced compared to the unimmunized controls. All animals (n=18) that became infected were euthanized in 2010; all uninfected animals (n=8) will be used in the next experiment. Experiment 2: Based on the data obtained from the challenge results in Experiment 1, 16 new Rhesus macaques and the 8 uninfected macaques from Experiment 1 are being used in this experiment. Animals will be vaccinated five times with either SIV DNA + Teck, Flu DNA +/- Meck or C.Diff DNA +/- Meck. After the last vaccination, all animals will be challenged with SIVsmE660. Blood and mucosal samples will be collected to monitor viral infection and immune response.