Study on tumor necrosis factor-producing cells induced by biological response modifiers.

OK-432在体外处理人外周血单核细胞，导致肿瘤坏死因子（TNF）、白细胞介素-1 α（IL-1 α）和白细胞介素-1 β（IL-1 β）的快速产生。用酶免疫分析（EIA）和免疫细胞化学染色鉴定新合成的TNF、IL-1 α和IL-1 β。虽然OK-432诱导细胞因子产生的能力各不相同，但上清液中IL-1 α和TNF活性在OK-432刺激后12小时达到最高水平。而IL-1 β活性则在6 h达到高峰，随后逐渐下降。在所检查的受试者中，观察到TNF和IL-1 α值之间的密切相关性（r=0.863）。在OK-432刺激后24小时，通过快速微波固定刺激的单个核细胞，使用特异性抗体免疫细胞化学证实了OK-432、TNF、IL-1 α和IL-1 β在细胞质中的积聚。TNF和/或IL-1的体外EIA可用于评估各种生物反应调节剂（BRM）诱导这些细胞因子产生的潜在能力，而微波固定技术对产生细胞因子的细胞进行免疫染色将有助于了解免疫反应的机制。

In vitro treatment of human peripheral blood mononulcar cells with OK-432 resulted in the rapid production or tumor necrosis factor (TNF), interlcukin-1alpha (IL-1alpha) and interlcukin-1beta (IL-1beta). The newly synthesized TNF, IL-1alpha and IL-1beta were identified by cnzyme immunoassay (EIA) and immunocytochemical staining. While the ability of OK-432 to induce to production of cytokines varied individually, IL-1alpha and TNF activities in supernatant reached maximal levels at 12 hours after OK-432 stimulation. In contrast, IL-1beta activity reached peak at 6 hours and then decreased gradually. Among the subjects examined, close correlations were observed between the value of TNF and IL-1alpha (r=0.863). TNF and IL-1beta (r=0.825), IL-1alpha and IL-1beta (r=0.986) at 24 hours after OK-432 stimulation, by rapid microwave fixation of stimulated mononuclear cells, cytoplasmic accumulation of OK-432, TNF, IL-Ualpha and IL-1beta was immunocytochemically demonstrated using specific antibodies. EIA of TNF and/or IL-1 in vitro will be useful for assessment of the potential ability of various biological response modifiers (BRMs) to induce the production of these cytokines, and immunostaining of the cells producing cytokines by microwave fixation technique will contribute to understand the mechanism of immuno responses.