Liu Wen-pei, Zheng Li-shu, Duan Zhao-jun, Xie Zhi-ping, Zhang Qian, Zhang Wan-ju, Hou Yun-de
State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Apr;23(2):100-2.
To construct human metapneumovirus (hMPV) DNA vaccines and evaluate the cellular and humoral immune response in mice.
Fusion protein FdeltaTM (without transmembrane domain) gene and M gene of hMPV were amplified from cDNA by PCR, then DNA vaccines pcDNA3.1His-FdeltaTM and pcDNA3.1His-M were constructed to verify the expression of F and M protein by Western blotting and indirect immunofluorescent assay (IFA) respectively. Serum IgG and spleen cell CTL were detected with ELISA and ELISPOT assay after the BALB/c mice were immunized intramuscularly with the vaccines.
The candidate DNA vaccines could express FdeltaTM and M protein as detected with Western blotting and IFA. The IgG antibody titers of mice was 1:44 when immunized with pcDNA3.1His-FdeltaTM, but could increase to 1:64 when co-immunized with pcDNA3.1His-M. ELISPOT assay demonstrated that IFN-gamma-secreting effector T cells reached 42 +/- 8.9 in co-immunization group, higher than single vaccine pcDNA3.1His-FdeltaTM group (32 +/- 7.4).
DNA vaccine pcDNA3.1His-FdeltaTM could induce specific cellular and humoral immune responses, and the immune response could increase when co-immunization with pcDNA3.1His-M was carried out.
构建人偏肺病毒（hMPV）DNA疫苗并评估其在小鼠体内的细胞免疫和体液免疫反应。
通过PCR从cDNA中扩增hMPV的融合蛋白FdeltaTM（无跨膜结构域）基因和M基因，然后构建DNA疫苗pcDNA3.1His-FdeltaTM和pcDNA3.1His-M，分别通过蛋白质免疫印迹法和间接免疫荧光法（IFA）验证F和M蛋白的表达。用疫苗对BALB/c小鼠进行肌肉注射免疫后，采用ELISA和ELISPOT法检测血清IgG和脾细胞CTL。
蛋白质免疫印迹法和IFA检测结果显示，候选DNA疫苗可表达FdeltaTM和M蛋白。用pcDNA3.1His-FdeltaTM免疫的小鼠IgG抗体效价为1:44，与pcDNA3.1His-M联合免疫时可增至1:64。ELISPOT法显示，联合免疫组分泌IFN-γ的效应T细胞达42±8.9，高于单一疫苗pcDNA3.1His-FdeltaTM组（32±7.4）。
DNA疫苗pcDNA3.1His-FdeltaTM可诱导特异性细胞免疫和体液免疫反应，与pcDNA3.1His-M联合免疫时免疫反应增强。
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