Manzel L, Macfarlane D E
Department of Medicine, VA Medical Center, and University of Iowa, Iowa City 52242, USA.
Life Sci. 1998;62(1):23-7. doi: 10.1016/s0024-3205(97)01034-5.
The mouse-mouse hybridoma plasma cell line 7TD1 requires exogenous IL-6 for growth, and has been used to characterize and assay IL-6. Oligodeoxynucleotides containing CpG (CpG-ODN) and bacterial DNA are immunostimulatory, preventing B-cell apoptosis and inducing cytokine synthesis. We report that the phosphorothioate CpG-ODN 5'-ATAATCGACGTTCAAGCAAG-3' (half maximal effect at 0.3 microg/ml) supports the growth of 7TD1 cells in the absence of added IL-6. A non-CpG control ODN was without effect. No IL-6 production by 7TD1 cells incubated with CpG-ODN was detected by sensitive immunoassay. CpG-ODN-supported growth was not influenced by IL-6-neutralizing antibody, but was completely abolished by chloroquine and quinacrine, agents which inhibit CpG-ODN responses in other cells. Our results show that CpG-ODN can substitute for IL-6 in this cell line. This cell line may be useful for assaying CpG-ODN activity and for screening congeners of chloroquine and quinacrine for their ability to inhibit CpG effects.
小鼠-小鼠杂交瘤浆细胞系7TD1的生长需要外源性白细胞介素-6(IL-6),并已用于鉴定和检测IL-6。含胞嘧啶-磷酸-鸟嘌呤(CpG)的寡脱氧核苷酸(CpG-ODN)和细菌DNA具有免疫刺激作用,可防止B细胞凋亡并诱导细胞因子合成。我们报告,硫代磷酸酯CpG-ODN 5'-ATAATCGACGTTCAAGCAAG-3'(在0.3微克/毫升时产生半数最大效应)在不添加IL-6的情况下支持7TD1细胞的生长。非CpG对照寡脱氧核苷酸没有作用。通过灵敏的免疫测定法未检测到与CpG-ODN孵育的7TD1细胞产生IL-6。CpG-ODN支持的生长不受IL-6中和抗体的影响,但被氯喹和奎纳克林完全消除,这两种药物可抑制其他细胞中的CpG-ODN反应。我们的结果表明,CpG-ODN可以替代该细胞系中的IL-6。该细胞系可能有助于检测CpG-ODN活性,并筛选氯喹和奎纳克林的同类物抑制CpG效应的能力。