鸭γ-干扰素活性体外检测方法的建立

Establishment of a method for detection of duck interferon - gamma bioactivity in vitro

目的：建立鸭γ－干扰素体外活性检测方法。方法：体外刺激巨噬细胞产生一氧化氮（ＮＯ）是γ－干扰素的生物学效应之一。为检测鸭 γ－干扰素活性，采用 Ｓｅｐｈａｄｅｘ Ｇ－１００诱导产生的腹腔巨噬细胞作为实验体系，观察一氧化氮浓度在ＬＰＳ，ＰＨＡ，Ｌ－精氨酸刺激的上述细胞中的变化情况。结果：结果表明上述方法制备的腹腔巨噬细胞经ＰＨＡ或ＬＰＳ活化后，细胞密度较高组一氧化氮产量明显升高，细胞密度较低组则没有显著差异。结论：高浓度的腹腔巨噬细胞对ＬＰＳ等刺激物敏感，可作为检测鸭γ－干扰素活性的重要手段。

Objective To establish a method for detection of duck interferon - gamma (DuIFN - γ) bioactivity in vitro. Methods One of the biological properties of IFN - γ is the ability of activating macrophage in vitro to generate nitric oxide (NO). To detect the duck interferon - gamma (DuIFN -γy) activity, we utilised duck peripheral blood mononuclear cells (PBMCs) and Sephadex G - 100 induced peritoneal exudate cells (PECs) as target cells to monitor the change of NO level of the target cells before or after stimulation of LPS, PHA of L - arginine. Results The results indicated that a limited increase of NO concentration following LPS or L - arginine stimulation was observed in chicken PBMCs,but not in duck PBMCs. The possible reason is assumed to be related to the differential distribution of macrophages in PBMCS from different species. Following stimulation with LPS or P~,the duck PECs with a high cell density produced a remarkable increase of NO concentration compared to the group with a low cell density. Conclusion Duck PEGs with high cell density are sensitive to the stimulation of LPS or P~,which can be used as a method to detect the activity of duck IFN - γ.