Using parenteral combination adjuvants to induce pan-mucosal cellular and humoral immunity

项目总结 大多数病原体通过粘膜表面进入人体，然而，迄今为止，大多数获得许可的疫苗都是注射的。 肠外，主要是肌肉内。虽然它们在激发系统免疫力方面表现出色，但并不总是如此。 诱导所需的粘膜免疫反应。这突显了我们对非粘膜组织如何 免疫可能被操纵以引起粘膜免疫反应，以及这种知识是如何 被利用来制造更好的针对粘膜病原体的疫苗。定义佐剂在这一过程中扮演的角色 反应是开发这种疫苗的关键；然而，决定佐剂驱动的粘膜的机制 抗体和细胞免疫反应还不是很清楚。我们已发表的工作和初步 使用主要组织相容性复合体I类(MHCI)和II类(MHCII)和B细胞四聚体进行实验 检测新型解毒细菌ADP皮内免疫后的粘膜免疫反应 核糖化肠毒素，称为dmLT，表明我们可以重定向内源性T和B细胞免疫 对肺、肠粘膜和女性生殖道(FRT)的反应。当dmLT与 安全、细菌来源的外膜囊泡(OMV)佐剂、CD4T细胞数量和疫苗特异性 抗体和B细胞甚至进一步增加。同时，OMV佐剂诱导显著扩张 疫苗特异性CD8 T细胞。此外，用dmLT或OMV佐剂的疫苗免疫可引起 保护肺部和肠道免受细菌感染。这些结果使我们假设皮内 DmLT+OMV联合免疫将抗原特异性B细胞和T细胞驱动到粘膜和 加强对黏膜病原体的疫苗保护。我们建议：1)确定皮内如何 DmLT与OMV佐剂联合免疫可将1)T细胞和2)B细胞定向到粘膜组织和 这些细胞是否是真正的组织驻留记忆细胞。同时，我们将检查免疫接种是否 使用dmLT-OMV佐剂疫苗可预防肺部、肠道的粘膜病毒和细菌感染 和FRT。这项研究将为佐剂如何调节粘膜和粘膜免疫提供新的见解。 允许我们引导反应，有利于消除病原体。

PROJECT SUMMARY Most pathogens enter the body at mucosal surfaces, yet, to date, the majority of licensed vaccines are injected parenterally, predominantly intramuscularly. While excellent at eliciting systemic immunity, they do not always induce the required mucosal immune responses. This highlights a gap in our understanding of how non-mucosal immunization might be manipulated to elicit mucosal immune responses and how such knowledge could be exploited to create better vaccines against mucosal pathogens. Defining the role that adjuvants play in this response is key to developing such vaccines; however, the mechanisms that dictate adjuvant driven mucosal antibody and cellular immune responses are not well understood. Our published work and preliminary experiments using major histocompatibility complex class I (MHCI) and II (MHCII) and B cell tetramers to examine mucosal immune responses after intradermal immunization with a novel detoxified bacterial ADP- ribosylating enterotoxin, called dmLT, demonstrate that we can retarget the endogenous T and B cell immune responses to the lung, intestinal mucosa, and female reproductive tract (FRT). When dmLT is combined with a safe, bacterial-derived outer membrane vesicle (OMV) adjuvant, CD4 T cell numbers and vaccine-specific antibodies and B cells are even further increased. Simultaneously, OMV adjuvant induces significant expansion of vaccine-specific CD8 T cells. Furthermore, immunization with dmLT- or OMV-adjuvanted vaccines elicits protection against bacterial infections in the lung and gut. These results lead us to hypothesize that intradermal immunization with combined dmLT plus OMVs will drive antigen-specific B cells and T cells to the mucosa and enhance vaccine protection against mucosal pathogens. We propose to: 1) determine how intradermal immunization with dmLT combined with OMV adjuvant directs 1) T cells and 2) B cells to mucosal tissue and whether these cells are bona fide tissue resident memory cells. In parallel, we will 3) examine if immunization with dmLT-OMV adjuvanted vaccines is protective against mucosal viral and bacterial infections in the lung, gut and FRT. This investigation will provide novel insights into how adjuvants regulate immunity at the mucosa and allow us to guide the response in favor of pathogen elimination.