Immune Response of Mosquitoes to Filarial Worms

蚊子对丝虫的免疫反应

• 批准号: 6659796
• 负责人: BRUCE MARTIN CHRISTENSEN
• 金额: $ 21.83万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-20 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6659796
• 关键词: Aedes; Culicidae; Filarioidae; arthropod borne communicable disease; arthropod genetics; arthropod nonpollutant control; biotechnology; cellular immunity; disease vectors; epizootiology; genetic library; genetically modified animals; host organism interaction; insect control; malaria; microarray technology; parasitism

DESCRIPTION (provided by applicant): The resurgence of mosquito-borne diseases is having a devastating impact on global health and an urgent need exists to develop new control strategies. A new strategy being actively investigated is the use of transgenics for the control of vector-borne diseases, i.e., to change vector competence by genetic transformation to reduce the ability of mosquitoes within a population to transmit a particular pathogen. However, if we are to evaluate the use of mosquito transformation as a viable disease control methodology it is imperative that we gain a better understanding of the mosquito's innate immune response that governs vector competence. Melanotic encapsulation is a cellular innate immune response that is responsible for the resistant phenotype of mosquitoes to filarial worms and malaria parasites. Through support from our parent grant (Immune Response of Mosquitoes to Filarial Worms) we are studying the biochemistry and genetic regulation of melanotic encapsulation, but during the course of these studies we have determined that detailed studies of gene expression in the circulating immune reactive cells (hemocytes) of mosquitoes is necessary if we are to clarify mechanisms of immune recognition and the initiation of defense responses. Very few hemocytes (<1,200) are present in an individual mosquito and they are not amenable to in vitro culture or even short-term in vitro maintenance; consequently, we have constructed cDNA libraries representing mRNA obtained from immune-activated hemocytes from two mosquito species. Generation of a limited number of ESTs from these libraries indicated a low percentage of ribosomal clones and revealed a number of matches to proteins with known innate immunity functions. These data convinced us that the systematic development of additional ESTs would facilitate the construction of informative microarrays that could be used to test specific hypotheses about the role hemocytes play in influencing vector competence. The budget of the parent grant was not designed to cover the costs required to generate the number of ESTs required, let alone the costs associated with oligonucleotide microarray construction; and, importantly, we could not have anticipated the feasibility of a comprehensive transcriptome approach to assess gene regulation in our model. The specific aims of the parent grant would be significantly enhanced by the multi-dimensional data from these additional tools. It is therefore appropriate to use the R21 mechanism for technology application to enhance our NIAID-funded research by (1) generating a large EST data set from immune-activated hemocyte cDNA libraries, (2) constructing oligonucleotide microarrays representing these ESTs, and (3) using these microarrays in data mining of hemocyte expression profiles to identify distinct patterns of transcription underlying the immune response of mosquitoes against filarial worms.

描述（由申请人提供）：蚊子传播疾病的复兴正在对全球健康产生毁灭性的影响，并且存在着制定新的控制策略的迫切需求。正在积极研究的一种新策略是使用转基因来控制媒介传播疾病，即通过遗传转化来改变向量的能力，以降低人群中蚊子传播特定病原体的能力。但是，如果我们要评估将蚊子转化作为一种​​可行的疾病控制方法的使用，那么我们必须更好地了解蚊子的先天免疫反应，从而控制媒介能力。黑素封装是一种细胞先天免疫反应，负责蚊子对丝状蠕虫和疟疾寄生虫的抗性表型。通过我们的父母赠款（蚊子对丝虫蠕虫的免疫反应）的支持，我们正在研究黑素封装的生物化学和遗传调节，但是在这些研究的过程中，我们确定了循环中的免疫反应性细胞（血细胞）的详细研究，如果我们的宣传是必不可少的，即使我们的宣传是必要的，即使宣传是必不可少的。单个蚊子中很少有血细胞（<1,200），它们不适合体外培养，甚至不适合短期维护；因此，我们构建了代表从两个蚊子物种的免疫激活血细胞获得的mRNA的cDNA文库。来自这些文库的数量有限的EST产生表明核糖体克隆的百分比较低，并揭示了与具有已知先天免疫功能的蛋白质的许多匹配。这些数据使我们确信，额外EST的系统开发将有助于构建信息丰富的微阵列，这些微阵列可用于测试有关血细胞在影响载体能力中起作用的特定假设。父母赠款的预算并非旨在支付产生所需的EST数量所需的费用，更不用说与寡核苷酸微阵列建设相关的成本了；而且，重要的是，我们无法预料到一种全面的转录组方法来评估模型中基因调节的可行性。这些其他工具的多维数据将大大提高父赠款的具体目标。 It is therefore appropriate to use the R21 mechanism for technology application to enhance our NIAID-funded research by (1) generating a large EST data set from immune-activated hemocyte cDNA libraries, (2) constructing oligonucleotide microarrays representing these ESTs, and (3) using these microarrays in data mining of hemocyte expression profiles to identify distinct patterns of transcription underlying the immune response of蚊子反对丝状蠕虫。