Ferraro A S, Newkirk M M
Department of Medicine, McGill University, Montreal General Hospital Research Institute, Quebec, Canada.
Hum Antibodies Hybridomas. 1993 Apr;4(2):80-5.
Peripheral blood B cells from normal individuals have been less than ideal as a resource for "fusible" cells for the generation of human hybridoma antibodies. In vitro stimulation of normal peripheral blood B cells by CD4+ T cells (HUT 78) that had been activated by a solid phase anti-CD3 monoclonal antibody (OKT3) was investigated to see if differentiation of B cells would result in an increased pool of B cells that could be immortalized. A comparison of the rate of successful fusion, an estimation of the frequency of the fusible cells from the input peripheral blood, and the amount of immunoglobulin secreted both in vitro, prior to fusion, and by the resulting clones in two different in vitro immunization protocols, with and without activated T cells indicated that inclusion of the activated T cells provided the necessary help to drive the B cells to a fusible state. Stimulation by activated T cells improves the efficiency of generating B cell hybrid clones from peripheral blood 10-fold compared to in vitro immunization with antigen alone.
作为用于生成人杂交瘤抗体的“可融合”细胞来源，正常个体的外周血B细胞一直不太理想。研究了用固相抗CD3单克隆抗体（OKT3）激活的CD4 + T细胞（HUT 78）对正常外周血B细胞的体外刺激，以观察B细胞的分化是否会导致可永生化的B细胞库增加。在两种不同的体外免疫方案中，有或没有激活的T细胞，比较成功融合的速率、输入外周血中可融合细胞频率的估计值，以及在融合前体外分泌的免疫球蛋白量和所得克隆分泌的免疫球蛋白量，结果表明加入激活的T细胞可提供必要的帮助，促使B细胞进入可融合状态。与仅用抗原进行体外免疫相比，激活的T细胞刺激可将从外周血生成B细胞杂交克隆的效率提高10倍。
