Novel ways to prevent upper GT infection

描述(申请人提供)：该项目的长期目标是确定重组拱顶纳米颗粒是如何产生生殖器粘膜辅助性T细胞1型(Th1)细胞的，这种重组拱顶纳米颗粒被设计成含有免疫原肽。几乎没有获得许可的疫苗用于预防粘膜组织的感染，这是因为我们缺乏对诱导粘膜免疫的疫苗设计的了解。我们假设，这些小型微生物大小的中空重组拱顶可以被设计成诱导粘膜免疫，并作为一种工具来识别关键的免疫触发因素，这些触发因素产生Th1细胞，并运输到生殖器粘膜组织。我们选择由病原体沙眼衣原体感染，它依赖Th1粘膜免疫反应来消除，是医疗保健的重大负担，目前还没有有效的疫苗。沙眼衣原体是性传播感染(STI)的主要原因，也是女性生殖功能障碍的煽动者，美国每年有100多万病例。小鼠适应的沙眼衣原体株经阴道分娩会引起类似于人类衣原体STI的性传播感染，并且可以通过阴道组织中存在Th1来预防。我们发现，与其他候选MOMP疫苗相比，用含有小鼠主要外膜蛋白的重组vault(MOMP-vault)免疫的小鼠表现出更好的抗攻击感染能力，并促进了T细胞向生殖器粘膜组织的迁移。在体外，MOMP-Vaults独立于已知的小鼠Toll样受体(TLRs)激活称为炎症体的复合体，并导致树突状细胞(DC)分泌选定的细胞因子。拱顶似乎通过溶酶体失稳来激活炎性小体，佐剂明矾也是如此。了解穹隆诱导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。在这项提案中，我们希望确定工程化穹隆如何刺激生殖器粘膜Th1免疫。我们将重点研究拱顶激活炎症小体的能力，炎症小体允许分泌促炎细胞因子，如白介素12(IL-12)，并在感染动物模型中确定拱顶诱导的炎症小体激活如何刺激生殖器粘膜Th1细胞免疫的产生。对拟议目标的调查将增进我们对开发针对衣原体STI的衣原体疫苗的了解，并有助于确定更有针对性的方法来设计针对其他感染黏膜组织的病原体的疫苗。 公共卫生相关性：粘膜免疫反应对传染病提供了更好的保护，但目前大多数获得许可的疫苗产生的是抗体反应，而不是Th1粘膜免疫，因为我们缺乏诱导粘膜免疫反应，特别是生殖器粘膜Th1细胞免疫的知识。沙眼衣原体可引起粘膜感染，是性传播感染(STI)的主要原因，美国每年有100多万病例。对由此导致的女性生殖功能障碍的治疗每年给美国医疗保健系统带来数十亿美元的负担，这项提案直接解决了确定如何诱导生殖器粘膜Th1细胞免疫以生产针对沙眼衣原体感染等感染的疫苗的问题。

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. PUBLIC HEALTH RELEVANCE: Mucosal immune responses provide superior protection against infectious disease, but most currently- licensed vaccines produce antibody responses instead of Th1 mucosal immunity because we lack knowledge for inducing mucosal immune responses, particularly genital mucosal Th1 cell immunity. Chlamydia trachomatis induces a mucosal infection and is a prominent cause of sexually transmitted infection (STI) with over 1 million cases in the U.S annually. Treatment for the resulting female reproductive dysfunction burdens the US health care system by billions of dollars annually and this proposal directly addresses the problem of identifying how to induce genital mucosal Th1 cell immunity for production of a vaccine against infections such as C. trachomatis STIs.