摘要
目的:探讨hCLCA1DNA疫苗对哮喘小鼠气道高反应性(AHR)的影响。方法:构建重组真核表达载体pSecTag-2A/hCLCA1,将其作为DNA疫苗肌注法接种给BALB/c小鼠(每2周1次),并采用间接ELISA法检测血清抗体。卵蛋白致敏法复制接种小鼠的哮喘模型。引喘5d后,测定小鼠气道压力峰值-时间指数(APTI)和离体气管环收缩最大张力,并计数肺泡灌洗液(BALF)中嗜酸性粒细胞的数量。设接种空载体pSecTag-2A对照组、生理盐水组和正常小鼠对照组。结果:间接ELISA法证实疫苗接种3次后,小鼠产生能结合mCLCA3胞外肽段的血清抗体,其滴度为1∶800-1∶1000。疫苗组、空质粒组和生理盐水组的APTI、气管环收缩张力和BALF中嗜酸性粒细胞含量均明显高于正常小鼠(P<0.01);但疫苗组的这些指标显著低于空载体组(P<0.01)。空载体组与生理盐水组结果相似。结论:人源hCLCA1基因疫苗能诱导小鼠产生结合自身mCLCA3的交叉抗体,并因此阻断了上皮对哮喘AHR的促进作用,从而在一定程度上抑制了哮喘AHR的发生。
AIM: To observe the preventive effect of DNA vaccine based on human calcium - activated chloride channel 1 (hCLCA1) on airway hyperresponsiveness in asthmatic mice. METHODS: The DNA vaccine was constructed by inserting the hCLCA1 gene into pSecTag - 2A, and then BALB/c mice were vaccinated by ira. once every two weeks. Serum antibody was checked with the antigen of mCLCA1 by ELISA analysis. Asthma was induced with ovalbumin in the vaccinated mice. The airway pressure time index ( APH), the contractile responsiveness of isolated tracheal rings and the number of eosinophil in bronchoalveolar lavage (BAL) were investigated. Mice injected with pSecTag - 2A, saline or normal mice were regarded as control groups. RESULTS: The title of antiserum binding to mCLCA1 in vaccine group was 1:800 to 1 : 1 000 after three times vaccination. Compared with normal group, APH, contractile responsiveness and number of eosinophil in vaccine group, pSecTag - 2A or saline group were increased markedly ( P 〈 0.01 ). These indexes in vaccine group were obviously lower than those in pSecTag - 2A and saline groups (P 〈 0.01 ). The latter two groups had the similar results. CONCLUSION: hCLCA1 DNA vaccine induces mouse to produce serum antibody binding to itself mCLCA3, and thus airway hyperresponsiveness and aggregation of eosinophil in asthmatic mouse are effectively inhibited.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2005年第9期1735-1738,共4页
Chinese Journal of Pathophysiology
基金
陕西省科学技术研究发展计划项目(No.2003K10G55)
西安市社会发展计划项目(No.SF200337)
关键词
哮喘
气道高反应件
疫苗
DNA
氯化物通道
钙激活
Asthma
Airway hyperresponsiveness
Vaccines, DNA
Chloride channels, calcium-activated