Continuous in vitro culture of Babesia microti

 描述（由申请方提供）：体外培养寄生虫的能力导致宿主-寄生虫关系、寄生虫生物学和病理生理学的理解快速进步。相反，缺乏寄生虫体外培养系统可能会严重阻碍对该特定病原体的研究，如在小巴贝斯虫引起的巴贝虫病中所见。尽管有越来越多的证据表明B的出现和传播，但这是事实。Microti在美国及其作为输血医学威胁的主导地位。我们的建议的总体目标是建立一个连续的体外培养系统的B。microti在人类红细胞，使一个平台适合未来的实验研究。众所周知，作为巴氏杆菌的细胞载体宿主的人RBC在循环中不是均匀、同质的细胞群，并且在其120天的寿命内经历许多与年龄相关的转化。我们的假设是，这些异质性红细胞是不平等的主机B。小孢子侵入和生长。我们提案中的两个具体目标将检验这一假设，并评估当呈现最佳年龄和特征的均匀RBC群体时，人体RBC中是否会发生寄生虫入侵和增殖。特定目标1旨在鉴定支持B最佳侵袭和复制的特定人RBC亚群。microti in vitro和Specific Aim 2将定义支持B的这种侵入和复制所需的培养条件（气体、培养基、振荡vs静态）。离体培养的Microti。通过结合我们在寄生虫学方面的丰富经验（Lobo和Narla，恶性疟原虫和B。divergens）以及红细胞专业知识（Narla）和试剂（B的人源。Microti、脐带细胞资源、抗体和抗原），我们将深入了解B偏爱的特定RBC谱系。显微镜这项研究是对寄生虫繁殖所需的培养条件的首次系统分析，也是寻找用于诊断、流行病学、治疗和预防人类巴贝西虫病的试剂以及协助开发筛选血液制品的新方法的必要的第一步。

 DESCRIPTION (provided by applicant): The ability to cultivate parasites in vitro has resulted in rapid progress in the understanding of host-parasite relationships, parasite biology and pathophysiology. Conversely, the lack of a parasite in vitro culture system can severely hamper studies on that specific pathogen as seen in Babesia microti-caused babesiosis. This is true despite the mounting evidence for the emergence and spread of B. microti in the US and its dominating status as a threat to transfusion medicine. The overall goal of our proposal is to establish a continuous in vitro culture system of B. microti in human RBCs to enable a platform amenable for future experimental investigation. It is well known that human RBCs which serve as the cellular vehicle host for Babesia are not a uniform, homogenous population of cells in circulation and undergo many age related transformations in their 120 day life span. Our hypothesis is that these heterogeneous RBCs are not equal hosts for B. microti invasion and growth. The two specific aims in our proposal will test this hypothesis and assess if parasite invasion and proliferation in human RBCs would occur when presented with a uniform RBC population of the optimum age and features. Specific Aim 1 addresses the identification of specific human RBC sub-populations that would support optimum invasion and replication of B. microti in vitro and Specific Aim 2 will define culture conditions (gas, media, shaking vs static) needed to support this invasion and replication of B. microti in vitro. By combining our extensive experience in parasitology (Lobo and Narla, P. falciparum and B. divergens) together with the red cell expertise (Narla) and reagents (Human sources of B. microti, umbilical cord cell resources, antibodies, and antigens) available at NYBC, we will obtain insights into the specific RBC lineage favored by B. microti. This study is the first systematic analysis of culture conditions required for parasite propagation and is a necessary first step in the search for reagents for diagnosis, epidemiology, treatment and prevention of human babesiosis, as well as to assist in developing novel methods to screen blood products.