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

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

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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）：蚊子传播疾病的重新出现已挑战科学界，以开发更有效的疾病控制，预防和治疗方法。为了做到这一点，必须在病原体生命周期的所有阶段对病原体宿主相互关系有更全面的了解。淋巴丝虫病是一种蚊子 - 传播疾病，感染了1.2亿人，再加上12亿人处于危险之中。这些线虫寄生虫与其蚊子载体具有的强制关系在某些蚊子中受到了损害，这些蚊子可以安装先天免疫反应。涉及黑色素在病原体上沉积的先天免疫反应是蚊子Armigeres subalbatus使用的自然抗药机制，以杀死人类丝状蠕虫病原体Brugia Malayi。但是，这种蚊子是密切相关的动物寄生虫Brugia Pahangi的有效向量。因此，AR。亚α-b。马来西-B。可以在实验室中轻松操纵的Pahangi系统，是表征先天免疫反应的理想模型，这些模型是蚊子源性丝虫病中载体能力的自然决定因素。我们以前的研究描述了这些防御反应中黑色素生物合成所需的生化途径，而新的分子工具和实验方法现在使我们能够开始揭示非自我识别的复杂性，靶标特异性黑色素沉积以及这种免疫反应所需的遗传调节/信号级数。在此，我们建议使用AR。亚α和丝状蠕虫B. Malayi，B。Pahangi和Dirofilaria Immitis介绍基因转录，并鉴定针对丝状蠕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6,000多个AR。亚α表达序列标签（EST）簇，以时间比较激活和控制蚊子之间的表达谱，并确定涉及此反应的关键因素。生物信息学，计算生物学和RNAI将用于评估涉嫌影响黑素化表型的基因，并确定在抗抗蠕虫免疫期间可能会共同调节的关键途径和其他因素。蛋白质组学方法（二维荧光差凝胶电泳和同位素编码的亲和力标签）具有基质辅助激光/飞行时间/质谱/质谱和液相色谱/tandem质谱法，可用于在响应和评估后的响应过程中进行临时介绍血液抗蛋白质组的时间，以评估疗程的变性。质谱法与亲和力隔离技术一致，将用于检验以下假设：模式识别分子与寄生虫上的不同分子相互作用，从而介导了与先知氧化酶蛋白络合物的进一步相互作用，该蛋白质的进一步相互作用，该复合物在预防性氧化酶的活化和墨拉兰人的产生中的生产中，在相互作用的位置上产生。公共卫生相关性：蚊子 - 传播的淋巴丝虫病提取了对人类健康的破坏性造成的损失，尤其是在世界热带地区的特权人群较低的人群中。该提案中描述的研究将使科学界更好地了解分子/遗传因素，这些分子/遗传因素控制蚊子和丝状蠕虫寄生虫之间兼容和不相容的关系。了解这些分子机制可以为可能用于控制蚊子传播丝虫病的新方法提供线索。