鸡白细胞介素-6的原核表达及其对疫苗免疫增强效果的研究  

Enhancment of immune respones to vaccines by recombinant chicken interleukin-6 (ChIL-6)

作　　者：谢彩华[1] 闫若潜[2] 崔保安[1] 赵雪丽[2] 吴志明[2] 张志凌[2] 张健[2] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]河南省动物疫病预防控制中心,河南郑州450008

出　　处：《中国预防兽医学报》2008年第7期557-561,共5页Chinese Journal of Preventive Veterinary Medicine

摘　　要：本研究采用PCR技术,扩增、克隆了鸡白细胞介素-6(ChIL-6)成熟肽基因并进行原核表达,对表达的鸡白细胞介素-6组蛋白(rChIL-6)通过尿素变性、复性液复性、PBS溶液透析等步骤进行纯化,经测定rChIL-6的促小鼠脾脏淋巴母细胞的增殖能力来评估重组蛋白的生物学活性;用4个不同浓度的rChIL-6在不同部位与NDV活疫苗、AIVH5N1灭活疫苗同时肌肉注射来评价其免疫增强作用。研究结果表明,表达rChIL-6为25ku左右,经纯化后的蛋白纯度在95%以上;100pg/mLrChIL-6注射组具有较高的促小鼠脾淋巴母细胞增殖活性;不同浓度的rChIL-6注射组对NDV和AIVH5N1疫苗都具有较好的免疫增强作用,但对其免疫抗体产生时间及抗体滴度峰值出现和维持时间均没有明显的影响,其中0.03μg/mLrChIL-6浓度注射组的对鸡疫苗的免疫增强作用最明显,过高或过低浓度的rChIL-6对疫苗免疫增强作用不显著。这些研究为新型免疫增强佐剂开发以及鸡免疫失败性疫病的预防和治疗奠定了基础。The chicken interleukin-6 （ChIL-6） gene was PCR amplified, sequenced and cloned into the prokaryotic expression vector pQE30. The expressed recombinant protein （rChIL-6） was purified under denaturing condition, refolding in renaturation buffer and dialysised. The bioactivity of rChIL-6 protein was estimated by testing its ability to stumulate the proliferation of IL-6- dependent lymphoblast cells from the spleen of mice. The immune enhancing effects of rChIL-6 protein was evaluated by injection of different concentrations of rChIL-6 protein and the Newcastle Disease Virus （NDV） attenuated live vaccine （La Sota strain）, avian influenza virus （AIV） H5N1 subtype oil-emulsion inactivated vaccine at the same time but at different parts of body. The result showed that injection of 100 pg/mL of ChIL6 could significantly promote the proliferation of lymphoblast cell from mice. The rChIL-6 exhibited enhanced immune protection induced by NDV and AIV H5N1 vaccines, but could not bring forward or prolong the antibody producing time, nor increase antibody titer. The injection of 0.03 μg/mL rChIL-6 exhibited the best effect of immune enhancement, neither higher nor lower concentration of rChIL-6 could promote the antibody titers elicited by vaccines.

关 键 词：鸡白细胞介素-6 基因克隆 蛋白表达 免疫活性 免疫增强 

分 类 号：S852.42[农业科学—基础兽医学]