Zaitsu Masafumi, Yamasaki Fumio, Ishii Eiichi, Midoro-Horiuti Terumi, Goldblum Randall M, Hamasaki Yuhei
Department of Pediatrics, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga City 849-8501, Japan.
Prostaglandins Leukot Essent Fatty Acids. 2006 Jan;74(1):61-6. doi: 10.1016/j.plefa.2005.09.005. Epub 2005 Nov 8.
The human basophilic cell line KU812 that is an established tool for studying the function of human basophils, is differentiated into mature basophils by interleukin (IL-3) or other agents. However, whether leukotrienes (LTs)-synthesis is affected by cytokines in KU812 cells remains unknown. KU812 cells were incubated with IL-3, IL-4, IL-6, IL-13 or IL-18 for up to 14 days. The A23187 stimulated- and IgE cross-linked-synthesis of LTC(4) and LTB(4) were measured using an enzyme immunoassay (EIA). The expression of messenger RNA (mRNA) for LT-synthesizing enzymes was examined by reverse transcriptase polymerase chain reaction (RT-PCR), and the expression of 5-lipoxygenase (5-LO) was examined by immunostaining. Incubation with IL-3 (10 ng/ml) and IL-18 (10 ng/ml) induced the expression of 5-LO. A23187stimulated LT-synthesis and IgE cross-linked LT-synthesis were enhanced after incubation with IL-3 or IL-18. These results indicated that IL-3 and IL-18 primed human basophils for higher LT-synthesis. Thus, both IL-3 and IL-18 might be important factors for regulating LT-synthesis during the differentiation of human basophils.
人嗜碱性粒细胞系KU812是研究人嗜碱性粒细胞功能的一种成熟工具,可通过白细胞介素(IL-3)或其他因子分化为成熟嗜碱性粒细胞。然而,白三烯(LTs)的合成在KU812细胞中是否受细胞因子影响仍不清楚。将KU812细胞与IL-3、IL-4、IL-6、IL-13或IL-18孵育长达14天。使用酶免疫测定法(EIA)测量A23187刺激的和IgE交联的LTC4和LTB4的合成。通过逆转录聚合酶链反应(RT-PCR)检测LT合成酶信使核糖核酸(mRNA)的表达,并通过免疫染色检测5-脂氧合酶(5-LO)的表达。用IL-3(10 ng/ml)和IL-18(10 ng/ml)孵育可诱导5-LO的表达。与IL-3或IL-18孵育后,A23187刺激的LT合成和IgE交联的LT合成增强。这些结果表明,IL-3和IL-18使人类嗜碱性粒细胞的LT合成增加。因此,IL-3和IL-18可能都是人类嗜碱性粒细胞分化过程中调节LT合成的重要因素。