DIFFERENTIATED CELL CULTURES: INFECTION BY MYCOPLASMAS

支原体是人类、实验室、家庭和农场的感染因子 动物、植物、昆虫和细胞培养物。 约15%的 连续的细胞系含有支原体。 几乎每一种细胞培养 参数受到影响。 支原体感染使结果无效， 体外诊断和实验程序，并浪费了数百万的联邦 每年提供美元。 随着新类型的 非成纤维细胞培养物在多种培养基中繁殖， 包括无血清制剂和大规模操作。 数据 迫切需要确定支原体的性质，在这个新的 细胞培养系统的产生。 本申请解决了某些 这些领域的细节：淋巴母细胞，细胞培养生长在 用于细胞大规模生长无血清培养基和检测方法 cultures. 细胞培养系统也可以代表有价值的模型进行研究 支原体生理学和发病机理。 支原体、脲原体和 螺原体在细胞培养基中不繁殖。 它们确实在生长， 然而，在细胞培养中，表明细胞产生肌浆细胞生长， 因素 这些研究将描述这些因素，并定义 机制，M。猪鼻病毒可以在T淋巴细胞系中生长， 无血清培养基中没有补充胆固醇，最近观察到， 这个实验室。 将对脲原体进行细胞和器官培养研究， 扩大 螺旋状质粒转化NIH 3T3细胞的研究 支原体，微小螺旋体，高效扩增， 希望能确定细胞和分子机制。 免疫结合 本实验室开发的用于鉴定细胞培养分离株的技术 提供了在临床标本中检测支原体的可能性。 这些 研究将扩大。 本提案所载的研究将产生重要信息 这将对细胞学专家有实际和直接的应用， 文化和传染病。 结果应有助于 改进检测方法，阐明支原体在 疾病，并更好地了解机制， 支原体-哺乳动物细胞关系，特别是那些发生在 体内和体外无血清条件。

Mycoplasmas are infective agents of humans, laboratory, domestic and farm animals, plants, insects and cell cultures. Approximately 15% of continuous cell lines contain mycoplasmas. Virtually every cell culture parameter is affected. Mycoplasmal infection invalidates results of in vitro diagnostic and experimental procedures and wastes millions of federal grant dollars annually. The problem will increase as new types of non-fibroblast cell cultures are being propagated in a variety of media, including serum-free formulations and in large scale operations. Data are critically needed to determine the nature of mycoplasmas in this new generation of cell culture systems. This application addresses certain of these areas in detail: lymphoblastoid cells, cell cultures grown in serum-free media and detection methods for large scale growth of cell cultures. Cell culture systems can also represent valuable models to study mycoplasmal physiology and pathogenesis. Mycoplasma, Ureaplasma and Spiroplasma species do not propagate in cell culture media. They do grow, however, in cell cultures, suggesting the cells produce myocplasma growth factors. These studies will characterize these factors as well as define mechanisms whereby M. hyorhinis can grow in a T lymphocyte cell line in serum-free media without supplemental cholesterol, recently observed in this laboratory. Cell and organ culture studies with ureaplasmas will be expanded. Studies on the transformation of NIH 3T3 cells by a helical mycoplasma, Spiroplasma mirum, with high efficiency will be expanded, hopefully to define cellular and molecular mechanisms. An immunobinding technique developed in this laboratory to identify cell culture isolates offers potential to detect mycoplasmas in clinical specimens. These studies will be expanded. Studies contained in this proposal will generate significant information that will have practical and immediate aplication to specialists in cell culture as well as infectious disease. Results should contribute to improved detection methods, elucidation of the role mycoplasmas play in disease, and to a better understanding of the mechanisms of mycoplasmal-mammalian cell relationships, especially those that occur in serum free condition in vivo and in vitro.