IMMUNIZATION OF ORAL MUCOSA FOR INDUCTION OF RECTAL AND GENITAL MOCOSAL 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。 像大多数其他性传播疾病（STD）一样，HIV的主要感染部位是粘膜表面，并且像大多数其他STD一样，没有针对HIV的疫苗。拟议的研究将探索通过普通粘膜免疫系统介导的远端部位免疫诱导，使用口腔粘膜途径进行艾滋病毒/性病免疫接种的可能性。 我们将描述在2个口腔粘膜部位（颊窝和舌扁桃体上的粘膜下免疫）接种疫苗后，外周和粘膜（口腔、鼻、肺和直肠）部位的先天性、适应性细胞和体液反应。 恒河猴将用一种 SIV版本的一个领先的艾滋病毒疫苗方案：DNA初免/复制缺陷的Ad 5疫苗加强。 我们将测试舌扁桃体上的粘膜下免疫是否产生强大的外周B和T细胞应答。将在血液和粘膜部位测量适应性应答;将使用ELISpot和细胞内细胞因子染色测量抗原特异性T细胞，并将通过ELISA测量抗体。将通过先天性免疫应答（全身树突状细胞表型和功能，以及血清细胞因子/趋化因子谱）研究不同途径引起的适应性应答差异的潜在机制。 本研究将提供关于口腔粘膜接种是否可以通过诱导由常见粘膜免疫系统介导的远端部位（如直肠）的免疫来提供针对性传播微生物的保护的数据。它还将研究这种免疫途径在需要鼻和肺免疫时对呼吸道病原体（如流感）的保护作用。 DNA引发在血液中引起低至不可检测的疫苗特异性细胞应答。然而，在支气管肺泡灌洗（BAL）高频率的多功能SIV特异性的CD 4+和CD 8 +T细胞检测ICS的DNA引发的IT动物，虽然DNA引发的IM动物表现出非常低的频率响应。IT组的CM 9 gag四聚体反应显著更高（P 0.005）。血浆中SIV特异性抗体应答在IT和IM组相似，但显示出不同的动力学特征。在用rAd 5加强后，应答显著增加; IT组在血液中产生相对较强的细胞应答，但各组的应答幅度差异无统计学显著性（%CM9+ CD 8+细胞; IT= 3.2%，IM=2.3%）。在IT组的粘膜部位观察到高水平的CD 8+应答（%CM9+ CD 8+细胞：BAL= 11.8%，结肠=1.5%）。 扁桃体免疫可以引起全身反应和远处的粘膜反应。粘膜反应的进一步表征，包括SIVmac 239对低剂量直肠攻击的保护，正在进行中。

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. Like most other sexually transmitted diseases (STDs), HIV's primary site of infection is at mucosal surfaces, and like most other STDs, there is no vaccine for HIV. The proposed study will explore the possible use of the oral mucosal route for immunization against HIV/STDs through induction of immunity at distal sites mediated via the common mucosal immune system. We will characterize innate, adaptive cellular, and humoral responses in the periphery and in mucosal (oral, nasal, pulmonary and rectal) sites to vaccines given at 2 oral mucosal sites (submucosal immunization in the buccal fossa and over the lingual tonsil). Rhesus macaques will be immunized with an SIV version of one of the leading HIV vaccine regimens: DNA prime/replication-incompetent Ad5 vaccine boost. We will test whether submucosal immunization over the lingual tonsil produces robust peripheral B and T cell responses. Adaptive responses will be measured in the blood and at mucosal sites; antigen-specific T cells will be measured using ELISpot and intracellular cytokine staining and antibody will be measured by ELISA. Possible mechanisms underlying differences in adaptive responses elicited by different routes will be investigated via studies of innate immune responses (systemic dendritic cell phenotype and function, and serum cytokine/chemokine profiles). This study will provide data on whether vaccination of the oral mucosa can provide protection against sexually transmitted organisms, through induction of immunity at distal sites (such as the rectum) mediated by the common mucosal immune system. It will also investigate the usefulness of this route of immunization for protection against respiratory pathogens, such as influenza, when immunity in the nose and lungs is desired. DNA priming elicited low to undetectable vaccine-specific cellular responses in blood. However, in bronchoalveolar lavage (BAL) high frequency polyfunctional SIV-specific CD4+and CD8+T cells were detected by ICS in DNA-primed IT animals, although DNA-primed IM animals showed very low frequency responses. CM9 gag tetramer responses were significantly higher in the IT group (P0.005). SIV-specific antibody responses in plasma were similar for IT and IM groups, but showed distinct kinetic profiles. Responses were significantly increased after boosting with rAd5; the IT group generated relatively stronger cellular responses in blood, but the difference in magnitude of responses by group was not statistically significant (%CM9+CD8+cells; IT=3.2%, IM=2.3%). High-level CD8+responses were seen at mucosal sites in the IT group (%CM9+CD8+cells: BAL=11.8%, colon=1.5%). Tonsillar immunization can elicit systemic responses and mucosal responses at distant sites. Further characterization of mucosal responses, including protection from low-dose rectal challenge with SIVmac239, is in progress.