摘要
目的制备鲍曼不动杆菌(Ab)A1S_1969重组蛋白,利用小鼠全身脓毒血症感染致死模型评价A1S_1969重组蛋白的免疫保护效果,探讨可能的免疫保护机制,为筛选Ab疫苗有效的保护性抗原奠定基础。方法基于反向疫苗学技术筛选出Ab外膜蛋白A1S_1969,利用p GEX-6P-2质粒构建GST融合表达载体,重组表达的A1S_1969蛋白经亲和层析高效纯化后与铝佐剂吸附制备而成重组A1S_1969免疫原;采用Balb/c小鼠全身感染模型评价A1S_1969重组蛋白的免疫保护效果,通过ELISA检测免疫小鼠Ig G抗体滴度和Ig G抗体亚型。制备A1S_1969重组蛋白抗血清进行体外调理吞噬杀菌实验。结果成功克隆、表达并纯化A1S_1969重组蛋白,纯度>90%。动物免疫保护结果显示A1S_1969重组蛋白组小鼠存活率为55.6%,对照组小鼠存活率为20.0%(P<0.05);末次免疫7 d后小鼠体内总Ig G抗体滴度为1∶64 000,以Ig G1亚型为主;体外调理吞噬杀菌实验结果显示A1S_1969特异性血清抗体可以增强中性粒细胞对Ab的杀菌作用,实验组杀菌率可达55%,对照组无杀菌活性。结论 A1S_1969重组蛋白可显著提高全身脓毒血症感染致死模型小鼠的存活率,具有良好的免疫保护效果。
In this study, we aimed to evaluate the immune protection of recombinant protein AIS_1969 from Acinetobacter baumannii on a systemic Acinetobacter infection mouse model. Outer membrane protein A1S_1969 gene from GenBank was cloned in pGEX-6p-2 plasmids, expressed in E.coli and purified by GST-affinity chromatography. Balb/c mice were immunized with recombinant A1S_1969 protein, and then challenged with Ab-17978 in 4.2× 108 CFU/ml through intraperitoneal injection. The mouse survival rate of A1S_1969 immunized group （56.6%） was significant higher than that of the control group （20%） （P〈0.05）. In addition, A1S_1969 recombinant protein immunization elicited a highly-titered IgG （1 ： 64 000） responses, mainly in IgG1 subclass, and the specific IgG antibodies enhanced bacterial elimination in mouse bloods. Taken together, A1S_1969 recombinant protein may protect mouse from lethal Ab infection and be worthy of further investigation as a candidate for Ab vaccine.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2016年第2期104-108,共5页
Immunological Journal
基金
国家重大科技专项(2013ZX09J13107-03B)
关键词
鲍曼不动杆菌
基因工程疫苗
克隆表达
重组蛋白
免疫保护
Acinetobacter baumannii
Recombinant vaccine
Cloning expression
Recombinant protein
Immune protection
