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目的 研究联合运用狂犬病毒糖蛋白基因重组腺病毒与同一抗原核酸疫苗对小鼠的免疫效果。方法 构建狂犬病毒糖蛋白 (GP)的真核表达质粒pcDNA3.1 CVS N2cGP作为核酸疫苗 ,瞬时转染COS 7细胞 ,间接免疫荧光试验检测pcDNA3.1 CVS N2cGP的表达 ;以基因枪方法进行初次免疫接种和首次加强免疫 ,以表达同一抗原的复制缺陷型重组腺病毒通过鼻腔接种进行第二次加强免疫 ;ELISA试验检测血清狂犬病毒特异性IgG抗体 ,快速荧光灶抑制试验 (RFFIT)检测狂犬病毒中和抗体。结果 间接免疫荧光试验表明pcDNA3.1 CVS N2cGP能有效地表达GP于转染的细胞膜上 ,ELISA试验表明小鼠接受基因枪核酸免疫后 ,仅诱导产生低水平的特异性抗体 ,而用重组腺病毒进行加强免疫后 ,特异性抗体水平显著提高。结论 联合运用狂犬病毒糖蛋白基因重组腺病毒与同种抗原核酸疫苗可克服它们单独使用时各自的缺点 ,能有效地诱导小鼠产生抗狂犬病毒特异免疫。
Objective To study the immune effect of combined use of rabies virus glycoprotein gene recombinant adenovirus and the same antigen nucleic acid vaccine on mice. Methods The eukaryotic expression plasmid pcDNA3.1 CVS N2cGP of rabies virus glycoprotein (GP) was constructed and transfected into COS 7 cells transiently. The expression of CVS N2cGP of pcDNA3.1 was detected by indirect immunofluorescence assay. The primary immunization with gene gun Vaccination and boosting for the first time to express the same antigen replication-deficient recombinant adenovirus nasal vaccination for a second boost; ELISA test for serum rabies virus-specific IgG antibodies, rapid fluorescence inhibition test (RFFIT) detection of rabies virus And antibodies. Results Indirect immunofluorescence assay showed that pcDNA3.1 CVS N2cGP could effectively express GP on the transfected cell membrane. ELISA test showed that mice immunized with gene gun only induced a low level of specific antibodies, whereas recombinant adenovirus After booster immunization, the level of specific antibodies increased significantly. Conclusions The combined use of rabies virus glycoprotein gene recombinant adenovirus and allogeneic antigen nucleic acid vaccine can overcome their shortcomings when used alone, and can effectively induce anti-rabies virus-specific immunity in mice.