Marker development for mosquito hemocytes

蚊子血细胞标记物开发

• 批准号: 6873553
• 负责人: Michael Strand
• 金额: $ 20.55万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6873553
• 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

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，天生能够识别多种外来入侵者。尽管过去几年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巨大进展，但人们对细胞防御反应的控制知之甚少。这在很大程度上是由于许多昆虫体型较小，这使得血细胞的收集和血细胞产生的效应分子的鉴定变得困难。通常也很难在体内对血细胞介导的防御反应进行操作性实验，或者分离特定的血细胞群用于体外实验。对于像蚊子这样的媒介节肢动物来说尤其如此，因为它们关于不同血细胞类型的起源、分化或功能的信息很少。蚊子血细胞主要根据形态进行分类，只有有限的数据将这些形态特征与功能特征相关。对于极其重要的载体，如冈比亚按蚊，几乎对任何生命阶段的血细胞个体发育一无所知。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。冈比亚基因组，以及我们对其他昆虫的血细胞谱系标记的了解，开发出一套强大的特征来识别冈比亚。冈比亚血细胞。鉴于该应用程序的探索性和 An 上可用的背景有限。冈比亚血细胞，我们认为将申请作为 R21 提案提交是适当的。具体目标是： 1. 克隆并表征 4a-3B 细胞亚群， 2. 表征 An 中的候选通用标记和基因特异性标记。冈比亚血细胞和 4a-3B 细胞，以及 3. 对亚克隆的 4a-3B 细胞进行蛋白质组分析，以确定新的候选标记。这项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重要工具。冈比亚蚊子和可能的其他种类的蚊子。