BIOCHEMISTRY OF INSECT HEMOCYTES

许多重要的人类寄生虫病是通过昆虫传播的。 向量。引起疟疾或丝虫病等疾病的生物体必须 躲避或击败昆虫媒介的防御反应，如果 成功地将病毒传播给人类宿主就可以了。然而， 昆虫与这种寄生虫之间的细胞和分子相互作用 生物体还没有被很好地理解。与我们详细的 脊椎动物免疫系统的知识，分子机制 昆虫的免疫反应在很大程度上仍然是一个谜。主要的反应是 昆虫对真核生物的寄生是对入侵生物的封装 通过寄主昆虫的血细胞。这个项目的长期目标是 鉴定和鉴定昆虫血细胞中的蛋白质 参与了封装过程。我们将通过以下方式解决这个问题 一组独特的抗Manduca血细胞的单抗的应用 我们已经生成的Sexta。具体目标是： 1)颗粒血细胞囊泡内分子的鉴定 在胞浆细胞和颗粒血细胞的表面参与 血细胞黏附和包膜。我们将对我们的单克隆组进行筛选 寻找那些可以扰乱浆细胞扩散、血细胞的抗体 体外试验中的粘连或包埋。 2)目标1中确定的分子的表征。我们将使用 作为试剂干扰血细胞反应的单抗 确定相应抗原的大小，以提纯 并克隆相应抗原的cDNA 从血细胞库中提取。我们将确定cDNA和DNA的序列 分析推导出的血细胞蛋白氨基酸序列。 鉴定在血细胞包膜中起作用的蛋白质是 这是了解它们如何运作的第一步，并最终走向 了解寄生虫如何避免昆虫的包囊反应 人类疾病的传播者。

A number of important human parasitic diseases are transmitted by insect vectors. Organisms that cause diseases such as malaria or filariasis must evade or defeat the defensive responses of their insect vectors if successful transmission to human hosts is to take place. However, the cellular and molecular interactions between insects and such parasitic organisms are not well understood. In comparison with our detailed knowledge of the vertebrate immune system, the molecular mechanisms of insect immune responses remain largely a mystery. The primary response of insects to eukaryotic parasites is encapsulation of the invading organism by the host insect's hemocytes. The long term objective of this project is to identify and characterize proteins from insect hemocytes that are involved in the encapsulation process. We will approach this problem with the use of a unique panel of monoclonal antibodies to hemocytes of Manduca sexta that we have generated. The specific aims are: 1) Identification of molecules within the vesicles of granular hemocytes or on the surface of plasmatocytes and granular hemocytes that are involved in hemocyte adhesion and encapsulation. We will screen our panel of monoclonal antibodies to find those that can disrupt plasmatocyte spreading, hemocyte adhesion, or encapsulation in in vitro assays. 2) Characterization of molecules identified in Aim 1. We will use the monoclonal antibodies that disrupt hemocyte responses as reagents to determine the size of the corresponding antigen, to purify the corresponding antigens, and to clone cDNAs for the corresponding antigens from hemocyte libraries. We will determine the sequences of the cDNAs and analyze the deduced amino acid sequences of the hemocyte proteins. The identification of proteins that function in hemocyte encapsulation is a first step toward understanding how they function and eventually toward understanding how parasites avoid the encapsulation response in insect vectors of human diseases.