ATTENUATED RECOMBINANT LISTERIA AS ORAL AIDS VACCINE

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。 我们首先确定了恒河猴中编码SIV gag的第三代减毒活单核细胞增生李斯特菌载体（称为Lmdd-BdopSIVgag）的最佳剂量。 接下来，我们检查了胃内疫苗接种的不同剂量方案。对照组接受“空”载体Lmdd-Bdop。 隔日接种不如连续三天每天接种。 为了测试先前用李斯特菌接种是否会耐受，我们用相同的Lmdd-BdopSIVgag以每天x3的时间表对已经经历完整疫苗接种系列的猴子进行再免疫;这组猴子被指定为组1B。 平行接种一组新的5只未处理猴（组1A）。 新的未处理对照（组2; 5只猴）仅接受空载体。 在组1A和1B中观察到良好的细胞免疫，有效地排除了耐受性。 然后两组均接受两次粘膜增强，其中活的编码SIV Gag的减毒腺病毒（Ad 5 hrSIVGag），这导致Gag特异性细胞免疫的强烈增加。 接下来，组1A和1B接受蛋白质免疫（三聚体HIV-1 gp 160和达特）以诱导中和抗体。 在蛋白加强结束时，用5个低的每周直肠内剂量的编码异源HIV进化枝C包膜的R5 SHIV攻击猴子。 观察到大量保护，包括预防感染。 我们的结论是，诱导平衡的免疫反应，包括细胞以及体液免疫可以成功。 用口服Lmdd-BdopSIVgag引发，随后用Ad 5 hrSIVGag粘膜加强诱导强细胞免疫，而用HIV-1 gp 160和达特产生交叉中和抗体。

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 first determined the optimal dose of the third-generation live attenuated Listeria monocytogenes vector encoding SIV gag, termed Lmdd-BdopSIVgag, in rhesus monkeys. Next, we examined different dose-schedules for intragastric vaccination. Controls received "empty" vector, Lmdd-Bdop. Every-other-day immunization was not as good as vaccinating daily for three consecutive days. To test whether prior vaccination with Listeria would tolerize, we re-immunized monkeys that had undergone a full vaccination series with the same Lmdd-BdopSIVgag with a daily x 3 schedule; this group of monkeys was designated Group 1B. A new group of 5 na¿ve monkeys was vaccinated in parallel (Group 1A). New na¿ve controls (Group 2; 5 monkeys) received only empty vector. Good cellular immunity was seen in Groups 1A and 1B, effectively ruling out tolerization. Both Groups then received two mucosal boosts with live attuenated adenovirus encoding SIV Gag (Ad5hrSIVGag), which resulted in strong increases in Gag-specific cellular immunity. Next, Groups 1A and 1B received protein immunizations (trimeric HIV-1 gp160 and Tat) to induce neutralizing antibodies. At the conclusion of the protein boosts, the monkeys were challenged with 5 low, weekly intrarectal doses of an R5 SHIV encoding a heterologous HIV clade C envelope. Substantial protection was seen, including prevention of infection. We conclude that inducing balanced immune responses consisting of cellular as well as humoral immunity can be successful. Strong cellular immunity was induced with oral Lmdd-BdopSIVgag priming, followed by mucosal boosting with Ad5hrSIVGag, whereas cross-neutralzing antibodies were generated with HIV-1 gp160 and Tat.