Development of a human T-T cell hybridoma secreting B cell growth factor-猫眼课题宝

分泌 B 细胞生长因子的人 T-T 细胞杂交瘤的开发

DOI：

发表时间：

1983

期刊：

Journal of Experimental Medicine

影响因子：

15.3

通讯作者：

中科院分区：

医学1区

文献类型：

作者：

J. L. Butler;A. Muraguchi;H. Lane;A. Fauci

正常人和鼠B细胞的长期培养的成功受到能够维持活化的B淋巴细胞增殖的可溶性因子的有限可用性的阻碍。以前在人体系统中使用各种培养物来源的上清液的实验不能通过免疫化学方法分离B细胞生长因子（BCGF）和白细胞介素2（IL-2）的活性。因此，在没有IL-2活性的情况下具有BCGF活性的纯化因子还不能用于研究。在本研究中，将正常人外周血T细胞与次黄嘌呤/氨基蝶呤/胸苷敏感的人T白血病细胞系CEM-6融合。在最近描述的人BCGF测定系统中，测试来自所得杂交细胞的上清液维持正常人B细胞增殖的能力。通过有限稀释克隆展示BCGF活性的杂交体。一个杂交克隆，2B 11，继续支持B细胞在长期培养和6天试验中的增殖，其水平显著高于常规生产的生长因子。杂种2B 11的上清中未发现IL-2活性。通过凝胶过滤分离杂交瘤上清液，并且B细胞的最大增殖由18- 20，000 mol wt蛋白质级分支持。因此，已经开发了在不存在任何可证实的IL-2活性的情况下具有BCGF活性的人T-T细胞杂交瘤。人T-T细胞杂交瘤分泌离散的免疫调节因子应被证明是有力的工具，在解剖人体淋巴细胞功能的免疫调节机制。

The success of long-term culture of normal human and murine B cells has been hampered by the limited availability of soluble factors capable of maintaining proliferation of activated B lymphocytes. Previous experiments using various culture-derived supernatants in a human system were unable to separate the activities of B cell growth factor (BCGF) and interleukin 2 (IL-2) by immunochemical means. Thus, purified factors with BCGF activity in the absence of IL-2 activity have not been available for study. In the present study, normal human peripheral blood T cells were fused with the hypoxanthine/aminopterin/thymidine- sensitive human T-leukemic cell line, CEM-6. Supernatants from the resulting hybrid cells were tested for the ability to maintain proliferation of normal human B cells in a recently described assay system for human BCGF. Hybrids demonstrating BCGF activity were cloned by limiting dilution. One hybrid clone, 2B11, continued to support proliferation of B cells in both long-term cultures and 6-d assays at a level significantly above that seen with conventionally produced growth factors. No IL-2 activity was found in the supernatant from hybrid 2B11. The hybridoma supernatant was fractionated by gel filtration, and maximum proliferation of B cells was supported by the 18-20,000 mol wt protein fraction. Thus, a human T-T cell hybridoma that has BCGF activity in the absence of any demonstrable IL-2 activity has been developed. Human T-T cell hybridomas secreting discrete immunoregulatory factors should prove to be powerful tools in dissecting the mechanisms of immunoregulation of human lymphocyte function.

Suppressor factor from a T cell hybrid inhibits delayed-type hypersensitivity responses to azobenzenearsonate.