Continuous in vitro culture of Babesia microti

田鼠巴贝斯虫体外连续培养

• 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
• 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

 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（由适用提供）：体外培养寄生虫的能力在理解宿主 - 寄生虫关系，寄生虫生物学和病理生理学方面取得了迅速的进展。相反，缺乏体外培养系统的寄生虫可能会严重阻碍有关该特定病原体的研究。这是真正的目的地，证明了Microti在美国的出现和传播及其主导地位作为对翻译医学的威胁。我们提案的总体目标是建立人类RBC中Microti的连续体外培养系统，以实现适合未来实验研究的平台。众所周知，作为贝贝西亚的细胞载体宿主的人类RBC并不是循环中的统一细胞群体，并且在其120天寿命中经历了许多与年龄相关的转变。我们的假设是，这些异质的RBC并不是小芽孢杆菌入侵和生长的宿主。我们的提案中的两个具体目标将检验该假设和评估，当在最佳年龄和特征的RBC人群中出现时，人类RBC的寄生虫入侵和增殖是否会发生。具体目的1解决了特定人类RBC亚群的鉴定，这些识别将支持在体外的最佳侵袭和复制，并且特定目标2将定义培养条件（气体，媒体，摇动与静态），以支持这种侵袭和复制Microti在体外。通过将我们在寄生虫学（Lobo和Narla，P。falciparum和B. divergens）和红细胞专业知识（NARLA）和试剂（Microti B. microti，脐带细胞资源，脐细胞资源，抗体和抗原的人体来源（人体来源）相结合，我们将在Nybc中获得互联网，B。这项研究是对寄生虫传播所需的培养条件进行的首次系统分析，是寻找诊断，流行病学，治疗和预防人类Babesiosis的试剂的必要第一步，并有助于开发筛查血液产品的新方法。