Cellular mechanisms by which Neisseria gonorrhoeae infects the female reproductive tract

项目摘要 性传播感染是一项重大的公共卫生挑战，也是一个严重的妇女健康问题， 因为妇女可能遭受这些感染的严重并发症：盆腔炎（PID），不孕症， 和易患危及生命的宫外孕然而，这些感染中的大多数是女性， 生殖道（FRT）无症状。FRT中的感染如何引起如此广泛的临床 在没有症状的情况下，结果仍然未知。了解非洲国家性传播感染的主要障碍是： FRT是缺乏一个模型，合理地模仿人类感染的所有方面。子宫颈是 第一反应时间内的性传播感染部位。宫颈粘膜不均匀，由多层非极化 外宫颈鳞状上皮细胞，内宫颈单层极化柱状细胞， 在转化区上皮细胞逐渐改变。虽然组织培养模型 对于解释特定的宿主-病原体相互作用，STI病原体如何处理不同的上皮细胞， 因为没有细胞培养模型可以模拟人类子宫颈的不同粘膜表面。 为了克服这些障碍，我们正在开发一种使用人类宫颈组织外植体的新感染模型 以实现我们的长期目标：阐明STI病原体感染FRT的机制。 为了实现这一目标，该建议侧重于淋病奈瑟菌（GC） 调节人类子宫颈的感染过程。GC导致淋病是第二常见的 由于多重耐药GC的激增，STI和全球公共卫生危机。了表层分子 的GC经历时相变化，这与其广泛的感染结果有关。我们假设 皮利和不透明相关蛋白（Opa）的不同变体的表达允许改变 在GC感染性，而人类宫颈上皮细胞的特性决定了哪些区域 很容易受到GC感染。为了验证这一假设，我们将使用我们的人类宫颈组织模型， 表达恒定Opas和皮利的GC的同基因菌株，以确定皮利和 Opa时相变化和宫颈上皮细胞的独特特性调节FRT中GC感染。 我们的宫颈外植模型打破了该领域的一个主要障碍，首次使其成为可能， 检查在GC感染至其体内靶向上皮细胞期间发生的细胞事件， 生理相关环境。这些研究将揭示新的机制，最终可以解释如何 GC根据其表面分子和上皮细胞的类型操纵信号传导和细胞骨架， 它们相互作用，使宫颈感染在定植性和穿透性之间转换。新组织 该项目建立的模型和感染机制可能从根本上改变我们追求 了解性传播感染，并为预防性传播感染的干预性药物设计开辟新的途径。

Project Summary Sexually transmitted infections (STIs) are a major public health challenge and a serious women's health issue, as women can suffer severe complications from these infections: pelvic inflammatory disease (PID), infertility, and predisposition to life-threatening ectopic pregnancy. However, the majority of these infections in the female reproductive tract (FRT) are asymptomatic. How infection in the FRT causes such a wide range of clinical outcomes in the absence of symptoms remains unknown. The primary obstacle to understanding STIs of the FRT is the lack of a model that reasonably mimics all aspects of human infection. The cervix is the initiation site for STIs in the FRT. The cervical mucosa is not uniform, composed of multilayered non-polarized squamous epithelial cells at the ectocervix, a single layer of polarized columnar cells at the endocervix, and the progressively changing epithelia in the transformation zone. While tissue culture models have contributed significantly to explaining specific host-pathogen interactions, how STI pathogens deal with different epithelia for infection is unclear, as no cell culture model can mimic the varying mucosal surfaces of the human cervix. To overcome these obstacles, we are developing a new infection model using human cervical tissue explants to address our long-term goal: to delineate the mechanisms by which STI pathogens infect the FRT. To pursue this goal, this proposal focuses on the cellular mechanism by which Neisseria gonorrhoeae (GC) modulates the infection process in the human cervix. GC causes gonorrhea that is the second most common STI and a public health crisis worldwide due to the upsurge of multi-drug resistant GC. The surface molecules of GC undergo phase variation, which has been implicated in its broad infection outcomes. We hypothesize that the expression of pili and distinct variants of opacity associated proteins (Opa) allows for changes in GC infectivity, while the properties of epithelial cells of the human cervix determine which regions are vulnerable to GC infections. To test the hypothesis, we will use our human cervical tissue model and isogenic strains of GC that express invariable Opas and pili to define the cellular mechanism by which pili and Opa phase variation and the distinct properties of cervical epithelial cells regulate GC infection in the FRT. Our cervical explant model breaks a major barrier of the field, making it possible for the first time to examine cellular events occurring during GC infection to their in vivo targeted epithelial cells under a physiologically relevant environment. These studies will reveal new mechanisms that can finally explain how GC manipulate signaling and cytoskeleton based on their surface molecules and the type of epithelial cells with which they interact to switch the cervical infection between colonizing and penetrating nature. The new tissue model and infection mechanisms established by this project may fundamentally change our way to pursue the understanding of STIs and open new avenues for interventive drug designs for the prevention of STIs.