Song Li-qiang, Li Yan, Qi Hao-wen, Liu Xin-ping, Wang Ji-cun
Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Zhonghua Yi Xue Za Zhi. 2004 Feb 17;84(4):329-33.
To observe the inhibitive effect on airway mucus overproduction with DNA vaccine based on human calcium-activated chloride channel 1 (hCLCA1) in asthmatic mice, who own mCLCA3 being xenogeneic homology of hCLCA1 in airway goblet cell.
The DNA vaccine was made with hCLCA1 gene inserted into pSecTag2B, and then BALB/c mice were vaccinated by i.m. once every two weeks. When serum antibody showed binding activity to mCLCA3 with ELISA analysis, asthma will be induced with ovalbumin in the vaccinated mice. To detect mucus production, lung sections were PAS stained and their MUC5AC mRNA levels were investigated by reverse transcription polymerase chain reaction (RT-PCR). Mice in control groups were injected with pSecTag2B/mCLCA3, pSecTag2B and saline, respectively.
Antiserum of vaccine group after three times vaccination showed good binding activity to three mCLCA3 extracellular domains (ED), and the activity to N-terminal C-terminal ED was stronger than middle-ED. After induced to asthma, the number of goblet cell and MUC5AC mRNA level in vaccine group were lower than these in control group.
hCLCA1 DNA vaccine can induce mouse to produce serum antibody binding itself mCLCA3, and thus airway mucus overproduction of asthmatic mouse is effectively inhibited.
观察基于人钙激活氯离子通道1（hCLCA1）的DNA疫苗对哮喘小鼠气道黏液过度分泌的抑制作用，该哮喘小鼠气道杯状细胞中存在与hCLCA1具有异种同源性的小鼠钙激活氯离子通道3（mCLCA3）。
构建携带hCLCA1基因的DNA疫苗并将其插入pSecTag2B载体，然后对BALB/c小鼠进行肌肉注射，每两周一次。当ELISA分析显示血清抗体对mCLCA3具有结合活性时，用卵清蛋白诱导接种疫苗的小鼠发生哮喘。通过过碘酸希夫（PAS）染色检测黏液分泌情况，并通过逆转录聚合酶链反应（RT-PCR）研究其黏蛋白5AC（MUC5AC）mRNA水平。对照组小鼠分别注射pSecTag2B/mCLCA3、pSecTag2B和生理盐水。
三次接种疫苗后疫苗组的抗血清对mCLCA3的三个细胞外结构域（ED）均显示出良好的结合活性，对N端和C端ED的活性强于中间ED。诱导哮喘后，疫苗组杯状细胞数量和MUC5AC mRNA水平低于对照组。
hCLCA1 DNA疫苗可诱导小鼠产生与自身mCLCA3结合的血清抗体，从而有效抑制哮喘小鼠气道黏液的过度分泌。
