Novel ways to prevent upper GT infection

描述（由申请人提供）：该项目的长期目标是确定重组拱顶纳米颗粒如何被改造成含有免疫原性肽，产生生殖器粘膜T辅助型1 （Th1）细胞。很少有许可的疫苗用于防止粘膜组织感染，即因为我们缺乏对设计诱导粘膜免疫的疫苗的理解。我们假设，小微生物大小的中空重组拱顶可以被设计成诱导粘膜免疫，并被用作识别产生Th1细胞的关键免疫触发因素的工具，Th1细胞可运输到生殖器粘膜组织。我们选择了病原体沙眼衣原体感染，它依赖于Th1粘膜免疫反应来消除，是卫生保健的重大负担，并且没有有效的疫苗。沙眼原体是性传播感染（STI）和女性生殖功能障碍的主要原因，在美国每年有超过100万例。阴道分娩小鼠适应沙眼衣原体（C. muridarum）可诱发与人类衣原体性传播感染相似的性传播感染，可通过阴道组织中存在Th1加以预防。我们发现，与其他MOMP候选疫苗相比，用含有C. muridarum主要外膜蛋白的重组vault （MOMP-vault）免疫的小鼠对攻击感染表现出更好的保护作用，并增强了T细胞向生殖器粘膜组织的迁移。在体外，momp -vault激活了称为炎性小体的复合物，独立于已知的小鼠toll样受体（TLRs），并引起树突状细胞（dc）分泌选择的细胞因子。穹窿似乎通过溶酶体的失稳激活炎症小体，就像佐剂明矾一样。了解穹窿诱导Th1生殖器粘膜免疫的机制将进一步发展对其他粘膜病原体的指导性免疫治疗。在这个提议中，我们希望确定工程拱顶如何刺激生殖器粘膜Th1免疫。我们将重点关注穹窿激活炎性小体的能力，它允许分泌促炎性细胞因子，如白细胞介素-12 (IL-12)，并确定在感染动物模型中，穹窿诱导的炎性小体激活如何刺激生殖器粘膜th1细胞免疫的产生。对提出的目标的研究将提高我们对开发针对衣原体性传播感染的衣原体疫苗的认识，并有助于确定针对感染粘膜组织的其他病原体设计疫苗的更有针对性的方法。

DESCRIPTION (provided by applicant): The long term goal of this project is to identify how recombinant vault nanoparticles, engineered to contain immunogenic peptides, generate genital mucosal T helper type 1 (Th1) cells. There are few licensed vaccines for protection against infections in mucosal tissues, namely because we lack an understanding for designing vaccines that induce mucosal immunity. We hypothesize that the hollow, recombinant vaults, the size of small microbes, could be engineered to induce mucosal immunity and be used as a tool to identify key immune triggers which produce Th1 cells that traffic to genital mucosal tissue. We chose infection by the pathogen, Chlamydia trachomatis, which relies on Th1 mucosal immune responses for elimination, is a significant burden on health care, and for which there is no effective vaccine. C. trachomatis is a prominent cause of sexually transmitted infection (STI) and instigator of female reproductive dysfunction, with over 1 million cases in the U.S. annually. Vaginal delivery of the mouse-adapted strain of C. trachomatis, C. muridarum, induces an STI similar to human chlamydial STI and can be prevented by the presence of Th1 within vaginal tissues. We found that mice immunized with recombinant vaults engineered to contain the major outer membrane protein of C. muridarum (MOMP-vault) demonstrated superior protection to a challenge infection as compared to other MOMP vaccine candidates and enhanced migration of T cells to genital mucosal tissues. In vitro, MOMP-vaults activated complexes called inflammasomes independently of known murine Toll-like receptors (TLRs) and caused the secretion of select cytokines from dendritic cells (DCs). Vaults appear to activate inflammasomes via lysosome destabilization as does the adjuvant, alum. Understanding the mechanisms whereby the vault induces Th1 genital mucosal immunity will further the development of instructive immunotherapy for other mucosal pathogens. In this proposal we wish to identify how engineered vaults stimulate genital mucosal Th1 immunity. We will focus on the ability of the vaults to activate inflammasomes, which allow secretion of pro- inflammatory cytokines such as interleukin-12 (IL-12), and determine how vault-induced inflammasome activation stimulates production of genital mucosal Th1-cell immunity in an animal model of infection. Investigations of the proposed aims will enhance our knowledge for developing chlamydial vaccines against Chlamydia STIs and help to define more targeted approaches for designing vaccines against other pathogens infecting mucosal tissues.