Verburg-van Kemenade B M, Weyts F A, Debets R, Flik G
Department of Experimental Animal Morphology and Cell Biology, Agricultural University, Wageningen, The Netherlands.
Dev Comp Immunol. 1995 Jan-Feb;19(1):59-70. doi: 10.1016/0145-305x(94)00047-j.
Carp, Cyprinus carpio L, macrophages and neutrophilic granulocytes obtained from pronephros were cultured. Supernatant was harvested after 48 h and tested for interleukin-1 (IL-1) bioactivity. A concentration-dependent stimulation of proliferation was found of carp Ig- lymphocytes as well as of the murine IL-1 dependent D10 (N4) M cell line. A 4 h treatment of cells with phorbol myristate acetate prior to culture gave a two- to fourfold enhancement of the bioactivity in the supernatant. Antibodies raised in sheep against human recombinant IL-1 alpha or IL-1 beta added to the supernatant annulled the IL-1 bioactivity. Western blot analysis of supernatants with sheep or rabbit polyclonal antisera against human IL-1s revealed 22 kDa and 15 kDa protein species. The predominant newly synthesized protein that was immunoprecipitated with these antisera was a 15 kDa molecular species. We conclude that carp macrophages and neutrophilic granulocytes produce an IL-1-like molecule with T-cell proliferating potency that shares structural similarities with mammalian IL-1. This is the first evidence for the IL-1 signal protein in carp immunocompetent cells.
对鲤（Cyprinus carpio L）的前肾巨噬细胞和嗜中性粒细胞进行培养。48小时后收集上清液并检测白细胞介素-1（IL-1）的生物活性。发现鲤Ig +淋巴细胞以及鼠IL-1依赖的D10（N4）M细胞系的增殖受到浓度依赖性刺激。在培养前用佛波醇肉豆蔻酸酯乙酸盐处理细胞4小时，使上清液中的生物活性提高了两到四倍。向上清液中加入羊抗人重组IL-1α或IL-1β产生的抗体可消除IL-1生物活性。用羊或兔抗人IL-1多克隆抗血清对上清液进行蛋白质免疫印迹分析，显示出22 kDa和15 kDa的蛋白质条带。用这些抗血清免疫沉淀的主要新合成蛋白质是一种15 kDa的分子条带。我们得出结论，鲤巨噬细胞和嗜中性粒细胞产生一种具有T细胞增殖能力的IL-1样分子，其与哺乳动物IL-1具有结构相似性。这是鲤免疫活性细胞中存在IL-1信号蛋白的首个证据。
