DIFFERENTIATED CELL CULTURES: INFECTION BY MYCOPLASMAS

支原体是人类、实验室、家庭和农场的感染源。 动物、植物、昆虫和细胞培养。约15%的 连续的细胞系含有支原体。几乎每一种细胞培养 参数受影响。支原体感染使In的结果无效 体外诊断和实验程序和浪费数百万联邦 每年提供助学金。随着新类型的 非成纤维细胞培养正在各种介质中繁殖， 包括无血清配方和大规模手术。数据是 急需确定这一新的支原体的性质 细胞培养体系的产生。这个应用程序解决了某些 这些方面的细节：淋巴母细胞，细胞培养在 细胞大规模生长的无血清培养基及检测方法 文化。 细胞培养系统也可以代表有价值的研究模型。 支原体的生理和发病机制。支原体、解脲支原体和 螺旋体不会在细胞培养液中繁殖。它们确实会生长， 然而，在细胞培养中，表明细胞产生了肌浆生长 各种因素。这些研究将描述这些因素的特征以及定义 猪鼻分枝杆菌在T淋巴细胞系中生长的机制 不含补充胆固醇的无血清介质，最近观察到 这个实验室。解脲支原体的细胞和器官培养研究将 扩大了。螺旋杆菌转化NIH3T3细胞的研究 支原体，螺旋体，米氏螺旋体，高效扩增， 希望能确定细胞和分子机制。一种免疫结合 本实验室开发的用于鉴定细胞培养分离物的技术 为检测临床标本中的支原体提供了可能性。这些 研究将会扩大。 这项提案中包含的研究将产生重要的信息 这将对细胞专家具有实用和即时的应用 文化以及传染病。结果应该有助于 改进检测方法，阐明支原体在 疾病，并更好地理解 支原体-哺乳动物细胞关系，尤其是发生在 体内和体外的无血清状态。

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.