Neisseria gonorrhoeae exploits host interferon epsilon to establish infection in the female urogenital tract

项目摘要 淋球菌(Ng)是一种性传播的革兰氏阴性细菌，可引起炎症和 盆腔炎(PID)。在美国，Ng的发病率正在急剧上升。令人担忧的是，吴昌俊 对所有批准的抗生素产生越来越多的抗药性。 干扰素是一种由女性泌尿生殖道上皮细胞高度表达的I型干扰素。 (在小鼠和人类中)，但在白细胞中不存在。与其他I型干扰素不同，干扰素-e不是由细菌诱导的。 “病原体相关分子模式”，如脂多糖(LPS)或核酸。相反，干扰素-e是 由泌尿生殖道中的性激素调节。我们发现雌激素通过以下途径促进Ng感染 诱导干扰素-e的表达。雌激素治疗显著延长女性生殖器的Ng感染 一条小路。I型干扰素，包括干扰素-e，共享一个共同的受体，即干扰素-α/β受体(IFNAR)。我们的数据 使用IFNAR和干扰素-e基因敲除(KO)动物，以及阻断针对IFNAR的单抗，强烈支持 雌激素诱导的I型干扰素导致Ng免疫逃避的假说。在没有IFNAR的情况下 作为信号，Ng实际上没有能力维持女性泌尿生殖道的定植。此外，当地的 给予重组干扰素-e(r干扰素-e)蛋白可完全逆转表型，使其与野生动物相似。 输入老鼠。初步研究表明，这可能与阳离子抗菌肽的调节有关。 (CAMP)，如抽筋，以及唾液酸化Ng脂低聚糖(LOS)。我们假设 雌激素诱导的干扰素-e是产生性Ng感染所必需的，因为它调节唾液酸的供应 Ng唾液酸基转移酶的前体，从而使Ng逃避AMP的杀伤。 在目标1中，我们将评估雌激素和干扰素-e对女性上皮细胞基因表达的影响。 在生殖道感染过程中使用小鼠和人类的Ng感染模型。我们将使用蛋白质组学和 确定干扰素-e是否调节cAMP和补体蛋白表达的基因表达方法 在生殖道里。在目标2中，我们将确定表达IFNAR的细胞类型如何促进Ng存活。我们会 还要评估I型干扰素对吞噬细胞内在杀菌活性的影响以及它们对 生殖道。在目标3中，我们将评估干扰素-e对ng基因，特别是调控基因的影响。 逃避营地，以及在没有干扰素-e的情况下对杀死Ng的影响。我们还将确定干扰素-e是否会减少 Ng对补体和/或通过调节唾液裂解的抽筋介导的杀戮的敏感性。 在性传播感染期间调节干扰素反应的能力是有效的和潜在的 改善或预防Ng感染和PID的破坏性后遗症的变革性策略。

PROJECT ABSTRACT Neisseria gonorrhoeae (Ng) is a sexually-transmitted gram-negative bacterium that causes inflammation and pelvic inflammatory disease (PID). In the U.S., the incidence of Ng is rising dramatically. Alarmingly, Ng has become increasingly resistant to all approved antibiotics. Interferon-epsilon (IFN-e) is a type I IFN that is highly expressed by epithelial cells of the female urogenital tract (both in mice and humans) but not in leukocytes. IFN-e, unlike the other type I IFNs, is not induced by bacterial “pathogen-associated molecular patterns,” such as lipopolysaccharides (LPS) or nucleic acids. Rather, IFN-e is regulated by sex hormones in the urogenital tract. We discovered that estrogen contributes to Ng infection by inducing the expression of IFN-e. Estrogen treatment dramatically prolongs Ng infection of the female genital tract. Type I IFNs, including IFN-e, share a common receptor, the IFN-alpha/beta receptor (IFNAR). Our data using IFNAR and IFN-e knockout (KO) animals, as well as blocking mAbs to IFNAR, strongly support the hypothesis that estrogen-induced type I IFNs contribute to Ng immune evasion. In the absence of IFNAR signaling, Ng is virtually incapable of maintaining colonization of the female urogenital tract. Furthermore, local administration of recombinant IFN-e (rIFN-e) protein completely reverted the phenotype to resemble the wild- type mice. Preliminary studies suggest that this may be related to regulation of cationic antimicrobial peptides (CAMP), such as CRAMP, as well as the sialylation of Ng lipooligosaccharide (LOS). We hypothesize that estrogen-induced IFN-e is required for productive Ng infection because it regulates the availability of sialic acid precursors to Ng sialyltransferase, thus, allowing Ng to evade AMP killing. In Aim 1, we will assess the impact of estrogen and IFN-e on gene expression in epithelial cells in the female genital tract during infection using both mouse and human models of Ng infection. We will use proteomics and gene expression approaches to determine if IFN-e regulates the expression of CAMPs and complement proteins in the genital tract. In Aim 2, we will determine how IFNAR-expressing cell types promote Ng survival. We will also assess the impact of type I IFN on the intrinsic bactericidal activity of phagocytes and their recruitment to the genital tract. In Aim 3, we will assess the impact of IFN-e on Ng genes, particularly genes that regulate CAMP evasion, and their impact on killing of Ng in the absence of IFN-e. We will also determine if IFN-e reduces the sensitivity of Ng to complement and/or CRAMP-mediated killing by regulating LOS sialyation. The ability to modulate IFN responses during sexually-transmitted infections is a valid and potentially transformative strategy to ameliorate or prevent the damaging sequelae of Ng infection and PID.