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

项目摘要 大多数病原体从粘膜表面进入人体，然而，迄今为止，大多数许可的疫苗都是注射的 胃肠外，主要是肌内。虽然它们在引发全身免疫方面很出色，但它们并不总是 诱导所需的粘膜免疫应答。这突出了我们对非粘膜上皮细胞如何 免疫可能被操纵以引起粘膜免疫应答，以及这些知识如何被 用来制造更好的疫苗对抗粘膜病原体。确定佐剂在这一过程中的作用 免疫应答是开发此类疫苗的关键;然而， 抗体和细胞免疫应答还没有被很好地理解。我们已发表的工作和初步的 使用主要组织相容性复合体I类（MHCI）和II类（MHCII）和B细胞四聚体的实验， 检查用新型解毒细菌ADP皮内免疫后的粘膜免疫应答， 核糖基化肠毒素，称为dmLT，证明我们可以重新靶向内源性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、细菌衍生的外膜囊泡（OMV）佐剂、CD 4 T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佐剂诱导显著扩增 疫苗特异性CD 8 T细胞。此外，用dmLT-或OMV-佐剂化疫苗免疫， 保护肺部和肠道免受细菌感染。这些结果使我们假设皮内 用组合的dmLT加OMV免疫将驱使抗原特异性B细胞和T细胞到达粘膜， 增强疫苗对粘膜病原体的保护。我们建议：1）确定皮内如何 用与OMV佐剂组合的dmLT免疫将1）T细胞和2）B细胞导向粘膜组织， 这些细胞是否是真正的组织驻留记忆细胞。与此同时，我们将检查免疫接种是否 与dmLT-OMV佐剂疫苗联合使用， 的FRT。这项研究将为佐剂如何调节粘膜免疫提供新的见解， 使我们能够引导反应有利于病原体的消除。

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.