ANAEROBIOSIS, CMP-NANA AND GONOCOCCAL PATHOGENESIS

厌氧、CMP-NANA 和淋球菌发病机制

• 批准号: 2068548
• 负责人: RICHARD REST
• 金额: $ 25.67万
• 依托单位: ALLEGHENY UNIVERSITY OF HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-06-01 至 1998-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2068548
• 关键词: N acetylneuraminate; Neisseria gonorrhoeae; anaerobiosis; antibacterial antibody; bacterial cytopathogenic effect; bacterial proteins; binding proteins; bioassay; cytosine nucleotides; enzyme activity; enzyme induction /repression; enzyme mechanism; gene expression; genetic library; gonorrhea; laboratory mouse; laboratory rabbit; lipopolysaccharides; method development; molecular cloning; nucleic acid sequence; nucleic acid structure; protein purification; sialyltransferases; site directed mutagenesis

Neisseria gonorrhoeae (the gonococcus, gonococci, gc) infects about 2 million civilians a year in the USA, and several millions more in the rest of the world. In women, untreated cervical disease (gonorrhea) often ascends to the endometrium and fallopian tubes to cause pelvic inflammatory disease (PID), which in turn can lead to fallopian tube scarring, sterility and ectopic pregnancy. Each year in the USA, treating gonorrhea, PID and their sequelae costs hundreds of millions of dollars. In addition to helping prevent and treat gonococcal infections and disease and their sequelae, studying gonococcal pathogenesis is important to help us better understand other mucosal pathogens, including those causing serious respiratory and meningeal infections (e.g., meningitis caused by N. meningitidis), and to help us better understand other sexually transmitted diseases. Gonococci grown in vitro interact with host defenses, including serum, epithelial cells and phagocytes, quite differently than do gc grown in vivo. The goal of the research presented in this proposal is to investigate the mechanisms by which gc modulate their lipooligosaccharide (LOS) under conditions thought to exist in vivo, i.e. in the presence of cytidine monophosphate N-acetylneuraminic acid (CMP-NANA) and under decreased oxygen tension (anaerobiosis). We have observed that anaerobiosis acts synergistically with low concentrations of CMP-NANA to induce high level serum resistance in gc. Serum resistance is due (at least in part) to the facile sialylation of LOS by gonococcal sialyltransferase (STase) using host CMP-NANA. We observed that anaerobic gc, compared to aerobic gc, express increased STase activity and altered LOS. To our knowledge, this is the first observation of the regulation of gonococcal virulence factors by conditions thought to exist in vivo (other than iron). To began to explain these observations, I propose the following SPECIFIC AIMS: 1. Develop a rapid, quantitative assay to measure gonococcal sialyltransferase (STase) activity. 2. Identify, clone, sequence and analyze the gonococcal STase gene(s), or genes associated with the expression of STase activity. 3. Purify and biochemically characterize the gonococcal protein(s) responsible for STase activity. 4. Begin investigations to determine the degree, level (transcriptional, translational, or post translational) and mechanism(s) of regulation of gonococcal STase activity or expression by anaerobiosis.

淋病奈瑟氏菌（Gonococcus，Gonococci，GC）感染约2 美国每年一百万平民，其余的数百万 世界。在女性中，未经治疗的宫颈疾病（淋病） 升上子宫内膜和输卵管引起骨盆 炎症性疾病（PID），进而导致输卵管 疤痕，不育和异位妊娠。每年在美国，治疗 Gonorrhea，Pid及其后遗症花费了数亿美元。 除了帮助预防和治疗淋球菌感染和疾病 和他们的后遗症，研究淋球菌发病机理对于帮助 我们更好地了解其他粘膜病原体，包括引起的粘膜病原体 严重的呼吸道和脑膜感染（例如，由 N. Meningitidis），为了帮助我们更好地了解其他性行为 传播疾病。 淋球菌在体外生长与宿主防御剂，包括血清， 上皮细胞和吞噬细胞，与生长的GC截然不同 体内。本提案中提出的研究的目的是 研究GC调节其脂肪糖酸的机制 （LOS）在被认为存在体内存在的条件下，即在存在的情况下 胞苷单磷酸N-乙酰神经氨酸（CMP-NANA）和下方 氧气张力降低（厌食症）。我们已经观察到 厌氧菌病以低浓度的CMP-NANA协同作用 在GC中诱导高水平的血清耐药性。血清耐药性应得 至少部分地是通过淋病杆菌轻松地靠LOS归还的。 使用宿主CMP-NANA使用siAllyltransferase（Stase）。我们观察到厌氧 与有氧GC相比，GC表达增加的基碱活性并改变了 洛斯。据我们所知，这是对监管的首次观察 淋球菌毒力因体内存在的条件（其他 比铁）。 为了开始解释这些观察，我提出了以下特定的特定观察 目的：1。开发快速的定量测定以测量淋球菌 辅助转移酶（Stase）活性。 2。识别，克隆，序列和 分析淋球菌基因基因或与该基因相关的基因 斯平活性的表达。 3。纯化和生化表征 负责骨活性的淋球菌蛋白。 4。开始 调查以确定程度，水平（转录， 翻译或后翻译后）和调节机制 通过厌食症的淋球菌活性或表达。