REACTIVE OXYGEN IN SNAIL/SCHISTOSOME INTERACTIONS

蜗牛/血吸虫相互作用中的活性氧

• 批准号: 2069203
• 负责人: VINCENT A. CONNORS
• 金额: $ 6.72万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2069203
• 关键词: Biomphalaria; SDS polyacrylamide gel electrophoresis; Schistosoma mansoni; cell mediated cytotoxicity; electrofocusing; enzyme linked immunosorbent assay; free radical oxygen; genetic strain; glycoproteins; high performance liquid chromatography; host organism interaction; hydrogen peroxide; interleukin 1; intracellular parasitism; ion exchange chromatography; laboratory mouse; larva; phagocytosis; protein sequence; schistosomiasis; superoxide dismutase

The long-term objective of this research is to elucidate the molecular, cellular, and biochemical mechanisms underlying cell-mediated cytotoxicity in strains of the snail, Biomphalaria glabrata, that are susceptible or resistant to infection with larval stages of the human blood fluke, Schistosoma mansoni. The system of study will be the NlH-Sm-PR-1 strain of S. mansoni and three strains of the snail intermediate host: the schistosome-susceptible M-line, and resistant 1O-R2 and 13-16-R1 strains of B. glabrata. Using this model a comparative functional, biochemical, and molecular approach will be used to determine if the reactive oxygen intermediates (ROIs), superoxide and hydrogen peroxide, are produced by snail hemocytes and if they are involved in snail hemocyte-mediated parasite killing. There are three specific aims to this research. The first aim is to determine the production and role of hemocyte-derived ROIs in larval schistosome killing. Baseline ROI levels will be quantified using an in vitro phagocytosis assay, in the presence or absence of snail plasma and"'or parasite excretory-secretory (E-S) products, while hemocyte ROI production during hemocyte-sporocyst contact will be determined using an in vitro encapsulation assay. ROI-mediated sporocyst killing will be determined using an established live-dead assay. The second aim is to isolate specific snail plasma and parasite E-S products that directly affect ROI production by identifying those products that effect hemocyte ROI production and phagocytosis separately. Plasma and E-S fractions will be obtained using molecular ultrafiltration. Components of fractions will be isolated using high performance liquid chromatography, ion exchange chromatography, and electrophoretic techniques. Effects of these components and fractions on hemocyte ROI production and killing of sporocysts will be determined using in vitro phagocytosis, live-dead, and encapsulation assays. The third aim is to isolate snail interleukin-1 and to further characterize a parasite-derived E-S antioxidant molecule. The function of both molecules will be evaluated for their influences on ROI production and effect during hemocyte encapsulation of the parasite. Peptide fragments from both molecules will be N-terminally sequenced in preparation for the future isolation and sequencing of genes associated with schistosome killing. This research will contribute significantly to the understanding of vector competence in schistosome-snail interactions, which should ultimately lead to novel and more efficient approaches to the control of schistosomes.

这项研究的长期目标是阐明分子， 细胞介导的细胞毒性的基础细胞和生化机制 在蜗牛的菌株中，易感性或 抗性感染的人类血液氟的感染， 曼氏菌。研究系统将是NLH-SM-PR-1菌株 S. Mansoni和蜗牛中间主持人的三种菌株： 具有螺旋体敏感的M线以及抗性1O-R2和13-16-R1菌株 B. glabrata。使用此模型比较功能，生化， 和分子方法将用于确定反应性氧是否是否 中间体（ROI），超氧化物和过氧化氢由 蜗牛血细胞，如果它们参与蜗牛血细胞介导 寄生虫杀死。这项研究有三个具体的目标。这 第一个目的是确定血细胞衍生的ROI的生产和作用 在幼虫方案中杀死。基线ROI水平将被量化 在存在或不存在蜗牛的情况下，使用体外吞噬作用测定 等离子体和“或寄生虫排泄物分泌（E-S）产物，而血细胞 血细胞 - 孢子胞菌接触期间的ROI产生将使用 体外封装测定法。 ROI介导的孢子囊杀死将是 使用已建立的死刑测定法确定。第二个目标是 隔离特定的蜗牛等离子体和寄生虫E-S产品 通过识别那些影响血细胞的产品来影响ROI的产生 ROI产生和吞噬作用分别。等离子体和E-S分数将 使用分子超滤获得。分数的组成部分将 使用高性能液相色谱分离，离子交换 色谱和电泳技术。这些影响 血细胞ROI生产和杀死的成分和分数 孢子囊肿将使用体外吞噬作用，死刑和 封装测定。第三个目的是隔离蜗牛白介素-1和 为了进一步表征寄生虫衍生的E-S抗氧化剂分子。 这 两个分子的功能将评估其对ROI的影响 寄生虫血细胞封装过程中的产生和作用。 来自两个分子的肽片段将在 为相关基因的未来隔离和测序做准备 与血吸虫杀死。这项研究将对 对矢量能力的理解，在螺旋体 - 螺旋相互作用中， 最终应该导致新颖，更有效的方法 控制细胞体的控制。

项目成果

Chemokinetic effect of interleukin-1 beta on cultured Biomphalaria glabrata embryonic cells.

IL-1β 对培养的光滑双脐胚胎细胞的化学动力学影响。

• DOI: 10.1645/ge-1867.1
• 发表时间: 2009
• 期刊: The Journal of parasitology
• 作者: Steelman,BrandonN;Connors,VincentA
• 通讯作者: Connors,VincentA