原核表达小熊猫IFN-γ的免疫活性研究

Study on Immune Activities of Prokaryotic Expression IFN-γ in Red Panda

为鉴定小熊猫IFN-γ原核表达产物的免疫活性,以纯化的小熊猫IFN-γ原核表达产物为材料,采用淋巴细胞增殖试验和ELISA,分别测定了IFN-γ对小熊猫淋巴细胞增殖及其分泌IFN-γ的影响。结果显示,与对照组相比,100μL浓度分别为10-1×1.56mg/mL、10-2×1.56mg/mL的纯化IFN-γ能够显著刺激淋巴细胞增殖(P<0.05);经热变性处理(煮沸10min)、浓度为10-1×1.56mg/mL的IFN-γ刺激淋巴细胞增殖的效果与空白对照无显著差异(P>0.05),但明显低于未处理组(P<0.05);淋巴细胞培养系统中加入浓度为10-1×1.56mg/mL的等体积IFN-γ,60h之内培养上清中的IFN-γ含量均显著高于对照组(P<0.01或P<0.05)。结果表明,原核表达的小熊猫IFN-γ产物具有刺激同种淋巴细胞增殖及分泌IFN-γ的活性。

To identify the activities of prokaryotic expression IFN-γ in red panda,lymphocyte proliferation test and ELISA were used to investigate the role of IFN-γon PBMC proliferation and IFN-γ secretion re-spectively.The results showed that PBMC proliferation could be dramatically enhanced by adding 100 μL purified IFN-γwith concentrations of 10-1 1.56 mg/mL and 10 ^-2 ×1.56 mg/mL compared with control （P 〈0.05）,while heat-denatured IFN-γwith a concentration of 10^ -1 ×1.56 mg/mL had no effect on PB-MC proliferation and no significant difference with control （P 〉0.05）.When culturing under purified IFN-γ stimulation at 10-1 1.56 mg/mL,the IFN-γ level in the supernatant was significantly higher than that of control within 60 h （P 〈0.01 or P 〈0.05）.In conclusion,the purified IFN-γcould stimulate the prolif-eration and IFN-γ secretion of homologous PBMC.