Development of a continous invitro culture system for Plasmodium vivax

间日疟原虫连续体外培养系统的开发

• 批准号: 7649325
• 负责人: Tetsuya Furuya
• 金额: $ 6.71万
• 依托单位: AMERICAN TYPE CULTURE COLLECTION
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-03 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7649325
• 关键词: Antimalarials; Aotus primate; Area; Biological; Biological Preservation; CD34 gene; Cell Fraction; Cell Line; Cells; Centers for Disease Control and Prevention (U.S.); Coculture Techniques; Competence; Development; Disease; Drug resistance; Erythrocytes; Erythropoiesis; Erythropoietin; Freezing; Future; Goals; Growth; Hematopoietic stem cells; Human; In Vitro; Infection; Interleukin-3; Laboratory Research; Length; Malaria; Malaria Vaccines; Mesenchymal; Methods; Monkeys; Mononuclear; Mus; Parasites; Patients; Plasmodium; Plasmodium falciparum; Plasmodium vivax; Prevention; Principal Investigator; Qualifying; Recombinants; Research; Research Personnel; Research Project Grants; Resistance development; Reticulocytes; Shipping; Ships; Site; Stem Cell Factor; Stem cells; Stromal Cells; System; Testing; Time; Umbilical Cord Blood; Vaccines; base; cytokine; drug development; novel; programs; public health relevance

DESCRIPTION (provided by applicant): The primary goal of this research project is to develop a continuous in vitro culture system for a human malaria parasite, Plasmodium vivax, which currently cannot be maintained in vitro for an extended period of time. The resulting system will be used for culturing P. vivax parasites from human patients to conduct important biological studies of P. vivax, such as drug resistance, and development of novel antimalarial drugs and vaccines. This will help with prevention and treatment of the disease caused by the parasite in endemic areas. To accomplish this goal, we will develop a system to obtain human reticulocytes, which is essential for P. vivax growth, by directing differentiation of the hematopoetic stem cells (HSCs) isolated from human umbilical cord blood (UCB), by using recombinant cell factors and co-culture with mouse or human stromal cells. The resulting reticulocytes will be infected with Plasmodium parasites (P. falciparum and P. vivax) to test if they can support growth of the parasites. The method for cryo- preservation of the cultured reticulocytes will also be studied for distribution of the reticulocytes to research laboratories and field sites. PUBLIC HEALTH RELEVANCE: The primary goal of this research project is to develop a continuous in vitro culture system of human reticulocytes for propagating a human malaria parasite, Plasmodium vivax. Currently, this system is not available and its absence impedes the development of drugs and/or vaccines for malaria caused by this parasite. To accomplish this goal, human reticulocytes will be developed through the directed differentiation of hematopoetic stem cells (HSCs) isolated from human umbilical cord blood (UCB). The enriched human reticulocytes will be tested for infection with Plasmodium parasites. Various methods for cryo- preservation of the cultured reticulocytes will be evaluated to create a qualified, long-term supply of the new culture system.

描述（由申请人提供）：该研究项目的主要目标是为人类疟原虫疟原虫Vivax开发一种连续的体外培养系统，目前无法长期维持体外维持。所得的系统将用于培养来自人类患者的维瓦克斯寄生虫，以对维瓦克斯进行重要的生物学研究，例如耐药性以及新型抗疟药和疫苗的发展。这将有助于预防和治疗流行地区寄生虫引起的疾病。为了实现这一目标，我们将开发一个系统，通过使用重组细胞因子和与小鼠或人类脊髓细胞共培养，通过指导从人脐带血（UCB）分离的造血干细胞（HSC）的分化，以获取人类的网状细胞，这对于维瓦克斯生长至关重要。所得的网状细胞将被寄生虫（恶性疟原虫和维瓦克斯疟原虫）感染，以测试它们是否可以支持寄生虫的生长。还将研究培养的网状细胞的冷冻保存方法，以分配网状细胞为研究实验室和现场部位的分布。 公共卫生相关性：该研究项目的主要目的是开发一种持续的人类网状细胞体外培养系统，用于传播人类疟疾寄生虫疟原虫。当前，该系统尚不可用，其缺席阻碍了该寄生虫引起的疟疾的药物和/或疫苗的开发。为了实现这一目标，将通过从人脐带血液（UCB）中分离出的造血干细胞（HSC）的定向分化来发展人类网状细胞。富集的人网状细胞将测试用疟原虫感染。将评估培养的网状细胞冷冻保存的各种方法，以创建合格的长期供应新培养系统。