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Marker development for mosquito hemocytes

蚊子血细胞标记物开发

基本信息

批准号：
7018523
负责人：
Michael Strand
金额：
$ 16.72万
依托单位：
UNIVERSITY OF GEORGIA
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-03-01 至 2008-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7018523
关键词：
Anopheles arthropod genetics cell differentiation cell line cell morphology cell population study cellular immunity disease vectors hemolymph immunogenetics immunologic receptors phagocytosis protein structure function proteomics

项目摘要

DESCRIPTION (provided by applicant): The insect immune system consists of both cellular and humoral elements that innately recognize broad classes of foreign intruders. While great progress has been made over the last several years in identifying humoral factors like antimicrobial peptides and the signaling pathways that regulate their synthesis, much less is known about control of cellular defense responses. This is due in large part to the small size of many insects which makes collection of hemocytes and identification of hemocyte-produced effector molecules difficult. It is also often difficult to conduct manipulative experiments on hemocyte-mediated defense responses in vivo or to isolate defined populations of hemocytes for use in experiments in vitro. This is particularly true for vector arthropods like mosquitoes where little information exists on the origins, differentiation or functions of different hemocyte types. Mosquito hemocytes are classified primarily on the basis of morphology with only limited data relating these morphological characters to functional features. In the case of critically important vectors, like Anopheles gambiae, virtually nothing is known about hemocyte ontogeny in any life stage. Here, our goal is to use hemocyte-like cell lines (4a-3B cells), the fully sequenced An. gambiae genome, and our knowledge of hemocyte lineage markers from other insects to develop a robust suite of characters for identifying An. gambiae hemocytes. Given the exploratory nature of the application and limited background available on An. gambiae hemocytes, we think it appropriate to submit the application as an R21 proposal. Specific aims are: 1. Clone and characterize subpopulations of 4a-3B cells, 2. Characterize candidate generic and gene specific markers in An. gambiae hemocytes and 4a-3B cells, and 3. Conduct a proteomic analysis of the subcloned 4a-3B cells to identify new candidate markers. The results of this study will provide essential tools for understanding the role of different hemocytes in cellular immunity of An. gambiae and likely other species of mosquitoes.

描述(由申请人提供)：昆虫免疫系统由细胞和体液成分组成，天生就能识别广泛类别的外来入侵者。尽管在过去的几年里，在识别抗菌肽等体液因子以及调节其合成的信号通路方面取得了很大进展，但对细胞防御反应的控制却知之甚少。这在很大程度上是由于许多昆虫的体积很小，这使得收集血细胞和识别血细胞产生的效应分子变得困难。在体内对血细胞介导的防御反应进行操纵性实验，或分离确定的血细胞群体用于体外实验，也往往是困难的。对于像蚊子这样的媒介节肢动物来说尤其如此，因为关于不同血细胞类型的起源、分化或功能的信息很少。蚊子血细胞的分类主要基于形态，只有有限的数据将这些形态特征与功能特征联系起来。就冈比亚按蚊等至关重要的媒介而言，几乎对任何生命阶段的血细胞个体发育都一无所知。在这里，我们的目标是使用血细胞样细胞系(4A-3B细胞)，即完全测序的AN。冈比亚的基因组，以及我们对其他昆虫的血细胞谱系标记的知识，以开发一套强大的特征来识别一个。冈比亚血细胞。考虑到该应用程序的探索性性质和可用背景有限，冈比亚血细胞，我们认为作为R21提案提交申请是合适的。具体目标是：1.克隆和鉴定4A-3B细胞亚群；2.鉴定AN中候选的通用标记和基因特异性标记。对亚克隆的4a-3B细胞进行蛋白质组学分析，寻找新的候选标记。本研究的结果将为了解不同血细胞在AN细胞免疫中的作用提供必要的工具。冈比亚亚科，可能还有其他种类的蚊子。