【摘要】 目的:探讨鸡γ-干扰素(IFN-γ)的最佳诱生条件。方法:采用夹心ELISA对培养的不同种类细胞在不同诱生剂作用下IFN-γ的分泌水平进行了检测,并对影响IFN-γ产量的条件进行了优化。结果:在ConA、PHA、ARV三种诱生剂中,ARV诱生能力最强,ConA其次,PHA最弱;其最佳诱生剂量分别为:ARV105TCID50/mL,ConA30μg/mL,PHA1.5μg/mL。鸡脾淋巴细胞、外周血白细胞(PBL)和鸡胚成纤维细胞(CEF)在同种诱生剂作用下,脾淋巴细胞IFN-γ分泌量最高,为47.65±3.26pg/mL。各种细胞最佳浓度均为3.0×106/mL。脾淋巴细胞在ConA和PHA刺激下,培养60h时产生的IFN-γ最高;而ARV诱导48hIFN-γ即可达峰值。同种IFN-γ具有启动效应。结论:确定了鸡IFN-γ的最佳诱生条件,为鸡IFN-γ定量检测奠定了基础。更多还原

【Abstract】 Objective:To explore the optimizing conditions of interferon-gamma(IFN-γ) induced.Method:The IFN-γ level induced by different inducers in different kind of cultured chicken cells was measured by Sandwich ELISA,and the factors influencing IFN-γ production were optimized.Result:ARV possessed the strongest IFN-γ inducing ability in three kinds of IFN-γ inducer(ConA,PHA and ARV),the other were ConA,PHA in proper order.The best inducing-dosage of ARV,ConA and PHA was 105 TCID50/ml,30 μg/ml and 1.5μg/ml respectively.In the supernatant of spleen lymphocyte,peripheral blood leukocyte(PBL) and chicken embryo fibroblast(CEF),the level of IFN-γ secretion was 47.65±3.26 pg/mL,was the highest in the spleen lymphocyte induced by the same inducer.The best cell consistence of different kind of cells were 3.0×106/ml.IFN-γ peaked in spleen lymphocyte stimulated by ConA and PHA at 60 h post-incubation,while ARV induced the highest IFN-γ level at 48 h post-incubation.Priming by the same species of IFN-γ were found in the IFN-γ induction with ARV.Conclusion:The optimizing conditions of IFN-γ induced were determined,that provided a base for quantitative detection of chicken IFN-γ.更多还原