葎草花粉主要变应原核酸疫苗pcDNA3.1-LC2的构建及其免疫原性鉴定  

Construction of humulus scandens pollen major allergen pcDNA3.1-LC2 DNA vaccine and study of its immunogenicity in mice

作　　者：张永红[1] 孙秀珍[1] 刘昀[1] 徐晶[1] 

机构地区：[1]西安交通大学医学院第二附属医院呼吸科,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2010年第1期51-53,78,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30772010)~~

摘　　要：目的构建葎草花粉主要变应原核酸疫苗pcDNA-LC2并鉴定其免疫原性。方法将pTripIEx2-LC2质粒用EcoRⅠ和Hind Ⅲ双酶切得到LC2葎草花粉变应原基因片段,将其插入pcDNA3.1(-)真核表达载体,在鉴定重组质粒构建成功后,大量制备去内毒素核酸疫苗pcDNA3.1-LC2,应用pcDNA3.1-LC2核酸疫苗免疫小鼠,取免疫血清行双向免疫扩散实验,分析其免疫原性。结果通过测序确认构建pcDNA3.1-LC2中的672 bp的编码框碱基与原序列一致,没有突变及缺失。应用pcDNA3.1-LC2免疫BALB/c小鼠,经双向免疫扩散试验验证pcDNA3.1-LC2在小鼠体内可以表达并产生特异性抗体。结论成功的构建了葎草花粉主要变应原核酸疫苗pcDNA3.1-LC2,pcDNA3.1-LC2可在小鼠体内表达,并能产生葎草花粉变应原特异性抗体,具有良好的免疫原性。Objective To construct humulus scandens pollen major allergen DNA vaccine pcDNA-LC2 and evaluate its immunogenicity. Methods Humulus scandens pollen allergen gene was digested from recombinant plasmid pTripIEx2-LC2 by EcoR Ⅰ and HindⅢ of restriction endonuclease, and then inserted into expression vector pcDNA3.1 （-）. A large amount of endotoxin pcDNA3. 1-LC2 purified plasmid was extracted after successful construction, and animal experiment was carried out to study its immunocompetence. Results Sequencing results showed that 672 bp in the coding sequence of frames of recombinant pcDNA3.1-LC2 was in line with the original base line, without deletion or mutation, pcDNA3.1-LC2 of pollen allergen humulus scandens allergen vaccinated BALB/c mice. Double immunodiffusion test showed that pcDNA3.1-LC2 could be expressed and produce antibodies in mice. Conclusion Humulus scandens grass pollen major allergen DNA vaccine pcDNA3. 1-LC2 has been successfully constructed. It can be expressed in mice, produces humulus scandens grass pollen allergen specific antibody and has a good immunogenicity.

关 键 词：支气管哮喘 葎草花粉变应原 核酸疫苗 

分 类 号：R562.25[医药卫生—呼吸系统]