Immune Response of Mosquitoes to Filarial Worms

蚊子对丝虫的免疫反应

基本信息

批准号：
7460491
负责人：
BRUCE MARTIN CHRISTENSEN
金额：
$ 39.25万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-04-01 至 2012-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7460491
关键词：
Address Affinity Affinity Chromatography Amino Acid Sequence Anabolism Animals Arthropods Asians Bacteria Biochemical Biochemical Pathway Biochemistry Bioinformatics Biomedical Research Brugia malayi Brugia pahangi Chorion Communities Compatible Competence Complement Complex Computational Biology Computer Simulation Computing Methodologies Culicidae Data Dependence Deposition Detection Dirofilaria immitis Encapsulated Evolution Expressed Sequence Tags Filarial Elephantiases Filariasis Fluorescence Future Gene Expression Gene Expression Profile Gene Expression Regulation Gene Proteins Gene Targeting Genes Genetic Genetic Transcription Glucans Goals Health Hemolymph Human Human Development Immune Immune response Immune system Immunity Insect Proteins Insecta Invertebrates Investigation Isotopically-Coded Affinity Tagging Knock-out Laboratories Lasers Life Cycle Stages Link Literature Mammals Mass Spectrum Analysis Mediating Melanins Methods Midgut Modeling Molecular Molecular Genetics Molecular Profiling Mosquito Control Mosquito-borne infectious disease Natural Immunity Natural Resistance Nature Nematoda Nucleotides Numbers Oligonucleotides Parasites Pathway interactions Pattern Pattern Recognition Personal Satisfaction Phase Phenotype Physiological Physiology Plasma Play Population Positioning Attribute Post-Translational Protein Processing Prevention Production Proteins Proteome Proteomics Public Health RNA Interference Reaction Regulation Relative (related person)Research Risk Role Sampling Signal Transduction Site Skin tanning System Techniques Testing Time Transcript Transcriptional Regulation Translations Wound Healing base cDNA Library comparative curdlan defense response disorder control egg enzyme substrate gel electrophoresis interest killings liquid chromatography mass spectrometry molecular mass novel strategies pathogen pro-phenoloxidase protein expression research study resistance mechanism response tandem mass spectrometry tool two-dimensional vector vector mosquito

项目摘要

DESCRIPTION (provided by applicant): The re-emergence of mosquito-borne diseases has challenged the scientific community to develop more effective means for disease control, prevention, and treatment. In order to do this, a more comprehensive understanding of pathogen-host interrelationships during all phases of the pathogen's life cycle must be developed. Lymphatic filariasis is a mosquito-borne disease infecting 120 million people and placing another 1.2 billion at risk. The obligate relationship these nematode parasites have with their mosquito vectors is compromised in certain mosquitoes that mount innate immune responses. Innate immune responses involving the deposition of melanin on the pathogen is the natural resistance mechanism employed by the mosquito Armigeres subalbatus to kill the human filarial worm pathogen, Brugia malayi. But, this mosquito serves as an effective vector for the closely related animal parasite, Brugia pahangi; consequently, the Ar. subalbatus-B. malayi-B. pahangi system, which can be easily manipulated in the laboratory, serves as an ideal model for characterizing innate immune responses that serve as natural determinants of vector competence in mosquito-borne filariasis. Our previous research deliniated the biochemical pathways required for melanin biosynthesis in these defense responses, and new molecular tools and experimental approaches now enable us to begin unravelling the complexities of non-self recognition, target specific melanin deposition, and the genetic regulation/signaling cascades required for this immune response. Herein we propose to use Ar. subalbatus, and the filarial worms B. malayi, B. pahangi and Dirofilaria immitis to profile gene transcription and identify distinct patterns underlying immune responses against filarial worms. These studies will employ oligo-nucleotide microarrays, representing over 6,000 Ar. subalbatus expressed sequence tag (EST) clusters, to temporally compare expression profiles between immune response activated and control mosquitoes and identify key factors involved in this response. Bioinformatics, computational biology, and RNAi will be used to assess genes suspected of influencing the melanization phenotype and to identify key pathways and other factors that might be co-regulated during anti-filarial worm immunity. Proteomics approaches (2-dimensional fluorescence difference gel electrophoresis and isotope-coded affinity tags) with matrix-assisted laser/time-of-flight/mass spectrometry and liquid chromatography/tandem mass spectrometry will be used to temporally profile the hemolymph proteome during responses and to assess post-translational modifications. Mass spectrometry, in concert with affinity isolation techniques, will be used to test the hypothesis that pattern recognition molecules interact with distinct molecules on parasites, mediating further interactions with prophenoloxidase-protein complexes that culminate in the activation of prophenoloxidase and the production of melanin at the site of interaction. PUBLIC HEALTH RELEVANCE: Mosquito-borne lymphatic filariasis extracts a devastating toll on human health, especially amongst less privileged populations in tropical regions of the world. The research described in this proposal will give the scientific community a better understanding of the molecular/genetic factors that control compatible and incompatible relationships between mosquitoes and filarial worm parasites. Understanding these molecular mechanisms could provide clues for new approaches that might be used in the control of mosquito-borne filariasis.

描述（由申请人提供）：蚊媒疾病的重新出现对科学界提出了挑战，要求他们开发更有效的疾病控制、预防和治疗手段。为了做到这一点，必须更全面地了解病原体生命周期所有阶段的病原体与宿主之间的相互关系。淋巴丝虫病是一种由蚊子传播的疾病，感染 1.2 亿人，并使另外 12 亿人面临风险。这些线虫寄生虫与其蚊媒之间的必然关系在某些产生先天免疫反应的蚊子中受到损害。涉及黑色素在病原体上沉积的先天免疫反应是蚊子用来杀死人类丝虫病原体马来丝虫的自然抵抗机制。但是，这种蚊子是密切相关的动物寄生虫彭亨布鲁虫的有效载体。因此，Ar。 subalbatus-B.马来-B. pahangi 系统可以在实验室中轻松操作，是表征先天免疫反应的理想模型，而先天免疫反应是蚊媒丝虫病媒介能力的自然决定因素。我们之前的研究阐明了这些防御反应中黑色素生物合成所需的生化途径，新的分子工具和实验方法现在使我们能够开始阐明非自我识别、目标特异性黑色素沉积以及所需的遗传调控/信号级联的复杂性对于这种免疫反应。在这里我们建议使用Ar。 subalbatus、丝虫马来丝虫、彭亨丝虫和恶丝虫来分析基因转录并识别针对丝虫的免疫反应的不同模式。这些研究将采用寡核苷酸微阵列，代表超过 6,000 个 Ar。 subalbatus 表达序列标签 (EST) 簇，用于临时比较免疫反应激活蚊子和对照蚊子之间的表达谱，并确定参与该反应的关键因素。生物信息学、计算生物学和 RNAi 将用于评估怀疑影响黑化表型的基因，并确定在抗丝虫免疫过程中可能共同调节的关键途径和其他因素。采用基质辅助激光/飞行时间/质谱和液相色谱/串联质谱的蛋白质组学方法（二维荧光差异凝胶电泳和同位素编码亲和标签）将用于在响应和分析过程中对血淋巴蛋白质组进行时间分析。评估翻译后修饰。质谱法与亲和分离技术相结合，将用于测试以下假设：模式识别分子与寄生虫上的不同分子相互作用，介导与酚氧化酶原-蛋白质复合物的进一步相互作用，最终导致酚氧化酶原的激活和黑色素的产生。互动的场所。公共卫生相关性：蚊媒淋巴丝虫病对人类健康造成毁灭性损害，尤其是世界热带地区的弱势群体。该提案中描述的研究将使科学界更好地了解控制蚊子和丝虫寄生虫之间相容和不相容关系的分子/遗传因素。了解这些分子机制可以为控制蚊媒丝虫病的新方法提供线索。