ATTENUATED RECOMBINANT LISTERIA AS ORAL AIDS VACCINE

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 我们首先确定了编码SIV Gag的第三代减毒单核细胞增多性李斯特菌载体Lmdd-BdoSIVgag在恒河猴体内的最佳剂量。接下来，我们检查了胃内接种的不同剂量计划。控件收到“空”向量Lmdd-Bdop。隔天免疫不如连续三天每天接种。为了测试之前接种李斯特菌疫苗是否可以耐受，我们用相同的Lmdd-BdoSIVgag重新免疫了接受了完整疫苗系列的猴子，每天×3个时间表；这组猴子被指定为1B组。一组新的5只NAéve猕猴被平行接种(1A组)。新的朴素对照(组2，5只猴子)只接受空载体。1A组和1B组细胞免疫功能良好，有效地排除了耐受性。然后，两组都接受了编码SIV Gag的活化腺病毒(Ad5hrSIVGag)的两次粘膜增强，这导致了Gag特异性细胞免疫的显著增加。 接下来，A组和1B组接受蛋白质免疫(三聚体HIV-1gp160和Tat)以诱导中和抗体。在蛋白质强化试验结束时，猴子们被要求每周经直肠注射5次低剂量的编码异源HIV分支C膜的R5Shiv。人们看到了实质性的保护措施，包括预防感染。我们的结论是，诱导由细胞免疫和体液免疫组成的平衡免疫反应是成功的。口服Lmdd-BdoSIVgag和Ad5hrSIVGag可诱导小鼠产生较强的细胞免疫应答，而HIV-1gp160和Tat可产生交叉中和抗体。

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.