Sarfati M, Rubio-Trujillo M, Wong K, Rector E, Sehon A H, Delespesse G
Immunology. 1984 Oct;53(2):187-96.
In view of the controversial data in the literature regarding the in vitro IgE synthesis by human lymphocytes, the conditions for culture of lymphocytes and the methodology for measurement of the IgE produced are described in detail. In the absence of any added mitogen, enriched B cell preparations derived from 70% of allergic donors actively secreted 100 to 3200 pg/ml of IgE after culture for 7 days, at which time the cell viability was higher than 85%. In comparable B cell cultures derived from non-allergic donors, only trace amounts of de novo synthesized IgE were detected in 20% of the cases. All B cell cultures actively secreted IgG, IgA, IgM and there was no apparent relationship between the secretion of IgE and that of the other classes of Ig. By contrast, the synthesis of IgE by unfractionated peripheral blood mononuclear cells of allergic individuals, which were stimulated with pokeweed mitogen (PWM) under several experimental conditions, was not consistently reproducible, i.e. the spontaneous synthesis of IgE in such cultures was either suppressed or enhanced by PWM. The most important finding was that the secretion of IgE was selectively enhanced by supplementing the B cell cultures with cell-free supernatants (CFS) of cultures of neonatal lymphocytes which had been preincubated with 10 micrograms/ml IgE. It is, therefore, concluded that B cell cultures from allergic individuals constitute an appropriate model for investigations of the mechanisms underlying the regulation of human IgE synthesis.
鉴于文献中关于人淋巴细胞体外合成IgE的数据存在争议,本文详细描述了淋巴细胞培养条件及所产生IgE的测量方法。在未添加任何促有丝分裂原的情况下,来自70%过敏供体的富集B细胞制剂在培养7天后可主动分泌100至3200 pg/ml的IgE,此时细胞活力高于85%。在来自非过敏供体的可比B细胞培养物中,仅在20%的病例中检测到微量的新生合成IgE。所有B细胞培养物均主动分泌IgG、IgA、IgM,且IgE分泌与其他类别的Ig分泌之间无明显关系。相比之下,在几种实验条件下用商陆有丝分裂原(PWM)刺激的过敏个体未分离外周血单核细胞合成IgE的情况并不一致,即此类培养物中IgE的自发合成被PWM抑制或增强。最重要的发现是,用10微克/毫升IgE预孵育的新生淋巴细胞培养物的无细胞上清液(CFS)补充B细胞培养物可选择性增强IgE的分泌。因此,得出结论,来自过敏个体的B细胞培养物构成了研究人类IgE合成调节机制的合适模型。