Lysozymes and immunity in Anopheles gambiae

冈比亚按蚊的溶菌酶和免疫

基本信息

批准号：
7470067
负责人：
SUSAN M PASKEWITZ
金额：
$ 24.5万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-09-30 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7470067
关键词：
Affect Africa Africa South of the Sahara Age Anopheles Genus Anopheles gambiae Anti-Bacterial Agents Antibodies Area Bacteria Binding Biological Assay Cessation of life Chemicals Child Competence Culicidae Development Disease Equilibrium Gene Expression Gene Silencing Genes Genetic Goals Human Human Resources Immunity Immunohistochemistry Infection Insecta Insecticide Resistance Knowledge Life Light Malaria Mammals Methods Microbe Midgut Molecular Mosquito Control Muramidase Numbers Oocysts Organism Parasite resistance Parasites Parasitic Diseases Pattern Pharmaceutical Preparations Plasmodium Plasmodium berghei Polymerase Chain Reaction Proteins Public Health Regulation Research Role Salivary Staging System Targeted Research Techniques Training Transcript Transgenic Organisms Western Blotting World Health Organization burden of illness cost design disability-adjusted life years disease transmission disorder control human disease in vivo interest killings novel strategies response statistics transmission process vector vector mosquito

项目摘要

DESCRIPTION (provided by applicant): Malaria, a disease caused by parasitic protozoans in the genus Plasmodium, is transmitted by mosquitoes to humans. Each year, hundreds of millions of people are infected with this organism and millions of fatalities result, mostly in young children. The problem is how worsening because of the increasing resistance of parasites to drugs. Mosquito control is also threatened because of insecticide resistance, increasing costs, and loss of trained personnel. Thus, new methods for malaria control are needed. An important target for research is the interaction between the mosquito and the parasite. The long-term goal of our research is to understand the molecular determinants of vector competence. We study the ways mosquitoes can kill parasites and how parasites avoid these responses as well as the mosquito factors that malaria parasites require for successful development. With this knowledge, we can attempt to enhance mechanisms that cause mosquito/parasite incompatibilitity through genetic or chemical manipulation of vector mosquitoes or to block the transmission of parasites by targeting mosquito molecules essential for parasite development. The proposed research will focus on how c-type lysozymes affect Plasmodium development in Anopheles gambiae. Silencing of the lysozyme c-1 gene results in a dramatic reduction in the number of mosquitoes that become infected with Plasmodium. Unexpectedly, Lys c-1 binds to oocysts, so this protein directly interacts with parasites. Specific Aim 1 will focus on 1) describing how Lys c-1 interacts with midgut and salivary stages of malaria parasites, using immunohistqchemical techniques; 2) determining whether anti Lys c-1 antibodies can be used to block parasite development; and 3) characterizing the contribution of Lys c-1 to antibacterial immunity, through gene silencing and subsequent infection with bacteria. Two other lysozymes, Lys c-2 and Lys c-7, are upregulated in the mosquito midgut when parasite infection occurs. Specific Aim 2 will examine 1) the effect of gene knockdown of Lys c-2 or c-7 on parasite development; and, 2) describing the changes in proteins and transcripts that occur during parasite infections, through western blotting, immunohistochemistry, and quantitative PCR.

描述（由申请人提供）：疟疾是一种由疟原虫属寄生原生动物引起的疾病，通过蚊子传播给人类。每年，数亿人感染这种微生物，导致数百万人死亡，其中大部分是幼儿。问题是由于寄生虫对药物的抵抗力不断增强，情况变得更加恶化。由于杀虫剂耐药性、成本增加以及训练有素的人员流失，蚊子控制也受到威胁。因此，需要新的疟疾控制方法。研究的一个重要目标是蚊子和寄生虫之间的相互作用。我们研究的长期目标是了解载体能力的分子决定因素。我们研究蚊子杀死寄生虫的方式、寄生虫如何避免这些反应以及疟疾寄生虫成功发育所需的蚊子因子。有了这些知识，我们可以尝试通过对媒介蚊子进行基因或化学操作来增强导致蚊子/寄生虫不相容的机制，或者通过针对寄生虫发育所必需的蚊子分子来阻止寄生虫的传播。拟议的研究将重点关注 c 型溶菌酶如何影响冈比亚按蚊的疟原虫发育。溶菌酶 c-1 基因的沉默会导致感染疟原虫的蚊子数量大幅减少。出乎意料的是，Lys c-1 与卵囊结合，因此该蛋白直接与寄生虫相互作用。具体目标 1 将重点关注 1) 使用免疫组织化学技术描述 Lys c-1 如何与疟疾寄生虫的中肠和唾液阶段相互作用； 2)确定抗Lys c-1抗体是否可用于阻止寄生虫发育； 3) 表征 Lys c-1 通过基因沉默和随后的细菌感染对抗菌免疫的贡献。当发生寄生虫感染时，另外两种溶菌酶 Lys c-2 和 Lys c-7 在蚊子中肠中表达上调。具体目标 2 将检查 1) Lys c-2 或 c-7 基因敲低对寄生虫发育的影响； 2) 通过蛋白质印迹、免疫组织化学和定量 PCR 描述寄生虫感染期间蛋白质和转录本的变化。