Butcher R D, McGarvie G M, Cushley W
Department of Biochemistry, University of Glasgow, U.K.
Immunology. 1990 Jan;69(1):57-64.
High-density human B lymphocytes were prepared from tonsillar mononuclear cells by depletion of adherent cells and T lymphocytes, followed by discontinuous density gradient centrifugation. The B cells were analysed for the levels of expression of the CD25 (Tac) antigen marker by flow cytometry following culture with a variety of cytokines. IL-4 could induce elevated levels of CD25 on high-density, putatively resting B lymphocytes in a dose-dependent fashion. Expression of CD25 at the B-cell surface could not be promoted by interleukin-2 (IL-2), interferon-gamma (IFN-gamma) or by a crude preparation of B-cell growth factor 2 (BCGF-2). Mitogenic challenge of the B cells with pokeweed mitogen (PWM) and a combination of phorbol ester and calcium ionophore were similarly ineffective, although a small increase in CD25 expression could be detected when the B cells were cultured with phytohaemagglutinin A (PHA). The ability of IL-4 to promote CD25 expression was abolished by the presence of IFN-gamma in the culture. Titration experiments suggested that the amount of IL-4 required to produce a half-maximal increase in CD25 expression was approximately 40 U/ml; this is considerably greater than the 8-10 U/ml required to produce the equivalent effect on CD23 expression. The ability of IL-4 to promote CD25 expression in the high-density B-lymphocyte population was apparently independent of proliferation of the cells. IL-4 could not promote Tac expression on high-density T cells prepared from the same tissue source.
通过去除贴壁细胞和T淋巴细胞,然后进行不连续密度梯度离心,从扁桃体单个核细胞中制备高密度人B淋巴细胞。在用多种细胞因子培养后,通过流式细胞术分析B细胞中CD25(Tac)抗原标志物的表达水平。IL-4能够以剂量依赖的方式诱导高密度、假定处于静止状态的B淋巴细胞上CD25水平升高。白细胞介素-2(IL-2)、干扰素-γ(IFN-γ)或B细胞生长因子2(BCGF-2)的粗制品均不能促进B细胞表面CD25的表达。用商陆有丝分裂原(PWM)以及佛波酯和钙离子载体的组合对B细胞进行促有丝分裂刺激同样无效,尽管当B细胞用植物血凝素A(PHA)培养时可检测到CD25表达有小幅增加。培养物中存在IFN-γ会消除IL-4促进CD25表达的能力。滴定实验表明,使CD25表达增加至最大值一半所需的IL-4量约为40 U/ml;这大大高于对CD23表达产生同等效应所需的8-10 U/ml。IL-4在高密度B淋巴细胞群体中促进CD25表达的能力显然与细胞增殖无关。IL-4不能促进从相同组织来源制备的高密度T细胞上Tac的表达。