Continuous in vitro culture of Babesia microti

田鼠巴贝斯虫体外连续培养

基本信息

批准号：
8961269
负责人：
Cheryl Ann Lobo
金额：
$ 25.71万
依托单位：
NEW YORK BLOOD CENTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8961269
关键词：
Address Age Agitation Antibodies Antigens Area Babesia Babesia microti Babesiosis Biological Biology Blood Blood Circulation Blood Screening Blood Transfusion Borrelia microti CD34 gene Carbon Dioxide Cattle Cell Aging Cells Centrifugation Coculture Techniques Culture Media Development Diagnosis Diagnostic Disease Ensure Environment Epidemiology Erythrocytes Erythroid Erythropoiesis Functional disorder Future Gases Goals Growth Heterogeneity Host-Parasite Relations Human In Vitro Individual Infection Intervention Invaded Investigation Longevity Medicine Methods Movement Mus Nutrient Optimum Populations Parasites Parasitology Phase Physical environment Plasmodium Plasmodium falciparum Population Prevention Protocols documentation Reagent Reporting Research Resources Reticulocytes Safety Serum Solutions Source Staging Supplementation Suspension substance Suspensions System Testing Ticks Transfusion Umbilical Cord Blood Umbilical cord structure United States Zoonoses age related blood product cell age density erythroid differentiation experience innovation insight interest monolayer neglect novel parasite invasion pathogen peripheral blood public health relevance safety study tissue culture transmission process

项目摘要

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.

描述（由申请人提供）：体外培养寄生虫的能力使人们对宿主-寄生虫关系、寄生虫生物学和病理生理学的理解取得了快速进展，缺乏寄生虫体外培养系统可能会严重阻碍这方面的研究。尽管有越来越多的证据表明田鼠巴贝斯虫在美国出现和传播，并且其对输血医学构成威胁，但这种情况确实存在。我们提案的总体目标是。在人红细胞中建立田鼠芽孢杆菌的连续体外培养系统，为未来的实验研究提供一个平台众所周知，作为巴贝虫细胞载体宿主的人红细胞并不是均匀、同质的细胞群。我们的假设是，这些异质红细胞并不是相同的宿主。我们提案中的两个具体目标将检验这一假设并评估寄生虫是否存在。当提供具有最佳年龄和特征的统一红细胞群时，人类红细胞就会发生入侵和增殖。具体目标 1 致力于识别特定的人红细胞亚群，以支持田鼠体外和特定的最佳入侵和复制。目标 2 将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 NYBC 提供的红细胞专业知识 (Narla) 和试剂（田鼠 B. microti 的人类来源、脐带细胞资源、抗体和抗原），我们将深入了解田鼠 B. microti 所青睐的特定红细胞谱系。这项研究是对寄生虫繁殖所需培养条件的首次系统分析，是寻找人类巴贝斯虫病诊断、流行病学、治疗和预防试剂以及协助开发试剂的必要的第一步。筛选血液制品的新方法。