Turner K J, Creany J, Coelen R J, Cameron K J, Holt B J, Beilharz M W
Department of Microbiology, University of Western Australia, Medical Centre.
Immunology. 1991 Dec;74(4):703-8.
The regulating effects of IL-4 and pokeweed mitogen on IgE synthesis in vitro by human peripheral blood leucocytes has been compared with the corresponding effect of these regulators on the expression of IgE mRNA. The latter was measured by dot blot hybridization with an oligonucleotide coding for a unique six amino acid region of the CH epsilon 2 domain. Specificity of the oligonucleotide probe was established by its inability to hybridize with RNA extracted from HMY-2 (IgG) and XQ-15 (IgM) secreting cell lines whilst producing intense signals with RNA extracted from the IgE secreting cell line U266. Whilst IgE mRNA was detected in RNA extracted from PBL of both atopic and control subjects, spontaneous IgE synthesis was restricted to atopic PBL. IL-4 increased both IgE mRNA and IgE synthesis in all PBL samples but PWM, while significantly increasing IgE mRNA expression either failed to modify IgE synthesis or actively suppressed it. The assay system employed to quantitate IgE synthesis in vitro was shown to be inhibited by both IgE binding factors and IgG anti-IgE autoantibodies which are produced in PBL cultures. IgE mRNA levels might therefore more accurately monitor the regulatory effects of IL-4 and PWM on IgE synthesis than quantitation of the IgE by radioimmunoassay.
已将白细胞介素-4（IL-4）和商陆有丝分裂原对人外周血白细胞体外IgE合成的调节作用，与这些调节剂对IgE mRNA表达的相应作用进行了比较。后者通过与编码CHε2结构域独特六氨基酸区域的寡核苷酸进行斑点印迹杂交来测定。寡核苷酸探针的特异性通过其不能与从分泌HMY-2（IgG）和XQ-15（IgM）的细胞系中提取的RNA杂交来确定，同时与从分泌IgE的细胞系U266中提取的RNA产生强烈信号。虽然在特应性和对照受试者的外周血淋巴细胞（PBL）提取的RNA中均检测到IgE mRNA，但自发IgE合成仅限于特应性PBL。IL-4增加了所有PBL样品中的IgE mRNA和IgE合成，但商陆有丝分裂原（PWM）虽然显著增加了IgE mRNA表达，但要么未能改变IgE合成，要么积极抑制它。用于体外定量IgE合成的检测系统被证明受到PBL培养物中产生的IgE结合因子和IgG抗IgE自身抗体的抑制。因此，与通过放射免疫测定法对IgE进行定量相比，IgE mRNA水平可能更准确地监测IL-4和PWM对IgE合成的调节作用。
