IMMUNE RESPONSES TO PLASMODIUM IN ANOPHELES GAMBIAE

冈比亚按蚊对疟原虫的免疫反应

• 批准号: 2073682
• 负责人: SUSAN M PASKEWITZ
• 金额: $ 15.54万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2073682
• 关键词: Culicidae; Plasmodium berghei; SDS polyacrylamide gel electrophoresis; binding proteins; disease vectors; genetic mapping; genetic strain; host organism interaction; immunity; laboratory mouse; laboratory rabbit; malaria; polymerase chain reaction

Malaria is the major vector-borne parasitic disease for much of the developing world and also threatens military personnel, tourists, business people and others who work in Africa, Asia and Latin America. Because of parasite and mosquito resistance to drugs and pesticides, new control strategies are needed. The long-term objective of the research is to understand the physiology, biochemistry, and genetics of mosquitoes that are resistant to malaria infections. This may suggest ways to replace natural populations with genetically transformed nonvector mosquitoes. A specific type of resistance is found in a genetically- selected strain of Anopheles gambiae which melanotically encapsulates ookinetes of many malaria species. A fully susceptible strain was also selected for comparison. The proposed research will focus on examining the biochemical and genetic basis for resistance. A newly-discovered model system of resistance that uses ion exchange beads will be used for genetic and biochemical studies. Plasmodium- resistant mosquitoes melanize negatively charged C-25 beads while susceptible mosquitoes do not. This system will greatly facilitate quantitative trait loci mapping of resistance genes, since no parasites need to be introduced and males can be scored. Markers to be used for mapping include an esterase locus which has been shown to be associated with resistance and microsatellite markers. A total of 137 microsatellite markers are now available from all autosomes and the X-chromosome. In addition to these mapping studies, the biochemistry of melanization in resistant mosquitoes and the lack of melanization in susceptible mosquitoes will also be examined. C-25 beads inoculated into susceptible mosquitoes and then dissected and introduced into resistant mosquitoes are not melanized. The nature of this protection will be studied by treatment of the beads with various enzymes prior to inoculation into resistant mosquitoes. SDS PAGE will be used to examine proteins on several types of beads destined to be melanized in the resistant strain and on unmelanized beads in the susceptible strain. Proteins associated with melanization or with the protective response in susceptible mosquitoes will be purified and used for polyclonal antibody production.

疟疾是许多国家主要的媒介传播寄生虫病 发展中国家，也威胁军事人员、游客、 商人和在非洲、亚洲和拉丁美洲工作的其他人。 由于寄生虫和蚊子对药物和杀虫剂具有抗药性，新 需要控制策略。研究的长期目标 是了解蚊子的生理学、生物化学和遗传学 对疟疾感染具有抵抗力。这可能会建议一些方法 用转基因非载体替代自然种群 蚊子。在基因上发现了一种特定类型的抵抗力 选定的冈比亚按蚊菌株，其包囊黑色素 许多疟疾物种的动动子。还发现了完全敏感的菌株 被选中进行比较。拟议的研究将侧重于检查 耐药性的生化和遗传基础。 新发现的使用离子交换的电阻模型系统 珠子将用于遗传和生化研究。疟原虫- 抗性蚊子使带负电的 C-25 珠子黑化，而 易受影响的蚊子则不会。该系统将极大地方便 抗性基因的数量性状基因座作图，因为没有寄生虫 需要引入并且可以对雄性进行评分。标记用于 作图包括已证明相关的酯酶位点 具有抗性和微卫星标记。共有137颗微卫星 现在所有常染色体和 X 染色体都可以使用标记。在 除了这些绘图研究之外，黑化的生物化学 抗性蚊子和易感蚊子缺乏黑色化 蚊子也将接受检查。 C-25珠接种到易感人群中 然后将蚊子解剖并引入具有抗药性的蚊子中 没有黑化。这种保护的性质将由 在接种之前用各种酶处理珠子 具有抵抗力的蚊子。 SDS PAGE 将用于检查蛋白质 几种类型的珠子注定会在抗性菌株中黑化 以及敏感菌株中未黑化的珠子上。相关蛋白质 与黑化或与易感者的保护反应 蚊子将被纯化并用于多克隆抗体生产。