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

淋病奈瑟菌利用宿主干扰素ε在女性泌尿生殖道中建立感染

• 批准号: 10435574
• 负责人: Douglas T Golenbock
• 金额: $ 79.61万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10435574
• 关键词: Affect; Anti-Bacterial Agents; Antibiotics; Antimicrobial Cationic Peptides; Attenuated; Bacteria; Bacterial Adhesion; Bacterial Infections; Bacterial Sexually Transmitted Diseases; Cells; Cervical; Complement; Cytidine Monophosphate N-Acetylneuraminic Acid; Cytometry; Data; Dichloromethylene Diphosphonate; Drug-resistant Neisseria Gonorrhoeae; Ectopic Pregnancy; Effector Cell; Environment; Enzyme-Linked Immunosorbent Assay; Epithelial; Epithelial Cells; Estrogen Therapy; Estrogens; Female; Female genitalia; Gene Expression; Genes; Gonadal Steroid Hormones; Gram-Negative Bacteria; HIV; Hormones; Host Defense; Human; Immune; Immune Evasion; Immune system; Incidence; Infection; Infertility; Inflammation; Innate Immune Response; Interferon-alpha; Interferons; Knock-out; Knockout Mice; Lead; Leukocytes; Lipid A; Lipopolysaccharides; Mediating; Menstrual cycle; Metabolism; Molecular; Monoclonal Antibodies; Mus; Myeloid Cells; Neisseria gonorrhoeae; Nucleic Acids; Pathway interactions; Pattern; Pelvic Inflammatory Disease; Phagocytes; Phase; Phenotype; Play; Predisposition; Production; Proteins; Proteomics; Public Health; Receptor Signaling; Recombinants; Regulation; Resistance; Role; Sexual Transmission; Sexually Transmitted Diseases; Sialic Acids; Sialyltransferases; Signal Transduction; Tissues; Vagina; Viral Proteins; Wild Type Mouse; Woman; antimicrobial; bactericide; cathelicidin antimicrobial peptide; cell type; chemokine; chronic pelvic pain; cytokine; efflux pump; enhancing factor; human model; in vivo; knockout animal; lipooligosaccharide; mouse model; neutralizing monoclonal antibodies; pathogenic bacteria; phosphoethanolamine; prevent; receptor; recruit; reproductive tract; response; sialic acid binding Ig-like lectin; sialylation; transcriptome sequencing; transmission process; type I interferon receptor; urogenital tract; virtual

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.

项目摘要 淋病奈瑟菌（Ng）是一种性传播的革兰氏阴性细菌，可引起炎症， 盆腔炎（pelvic inflammatory disease，PID）在美国，Ng的发病率正在急剧上升。令人担忧的是，Ng 对所有被批准的抗生素的耐药性越来越强。 干扰素-γ（IFN-γ）是一种I型干扰素，由女性泌尿生殖道上皮细胞高度表达 (both在小鼠和人中）但不在白细胞中。与其他I型IFN不同，IFN-e不受细菌诱导， 病原体相关的分子模式，如脂多糖（LPS）或核酸。相反，IFN-e是 由泌尿生殖道的性激素调节。我们发现雌激素通过以下方式促进Ng感染： 诱导IFN-γ的表达。雌激素治疗显著降低女性生殖器感染 道。I型IFN，包括IFN-ε，共享共同的受体，IFN-α/β受体（IFNAR）。我们的数据 使用IFNAR和IFN-e敲除（KO）动物，以及阻断IFNAR的mAb，强烈支持 雌激素诱导的I型IFN有助于Ng免疫逃避的假设。如果没有IFNAR 由于Ng信号转导，Ng实际上不能维持女性泌尿生殖道的定殖。此外，当地 给予重组IFN-ε（rIFN-ε）蛋白完全恢复了表型，与野生型相似。 型小鼠。初步研究表明，这可能与阳离子抗菌肽的调控有关 在一些实施方案中，Ng脂低聚糖（LOS）的唾液酸化包括CAMP（如CRAMP）的唾液酸化，以及Ng脂低聚糖（LOS）的唾液酸化。我们假设 雌激素诱导的IFN-e是生产性Ng感染所必需的，因为它调节唾液酸的可用性 Ng唾液酸转移酶的前体，从而允许Ng逃避AMP杀伤。 在目的1中，我们将评估雌激素和IFN-e对女性卵巢上皮细胞基因表达的影响。 使用Ng感染的小鼠和人模型在感染期间的生殖道。我们将使用蛋白质组学， 基因表达方法，以确定IFN-e是否调节CAMPs和补体蛋白的表达 在生殖道中。在目标2中，我们将确定IFNAR表达细胞类型如何促进Ng存活。我们将 还评估了I型IFN对吞噬细胞内在杀菌活性的影响，以及吞噬细胞的募集， 生殖道在目标3中，我们将评估IFN-e对Ng基因的影响，特别是调节Ng基因表达的基因。 CAMP逃避，及其在缺乏IFN-e的情况下对Ng死亡的影响。我们还将确定IFN-e是否减少 Ng对补体和/或通过调节LOS唾液酸化的CRAMP介导的杀伤的敏感性。 在性传播感染期间调节IFN应答的能力是一种有效的和潜在的治疗方法。 这是一种改善或预防Ng感染和PID的破坏性后遗症的变革性策略。