牙龈卟啉单胞菌基因疫苗pcDNA3.1(+)/kgpcd免疫原性研究

目的:研究重组质粒pcDNA3. 1( ) /kgpcd免疫原性,并观察目的基因编码的蛋白在小鼠肌肉及颌下腺组织中的原位表达情况。方法:重组质粒pcDNA3. 1 ( ) /kgpcd经肌肉注射和颌下腺区注射免疫BALB/c小鼠,采用间接ELISA方法检测免疫后BALB/c小鼠血清中IgG和唾液中sIgA抗体水平;用免疫组织化学染色观察目的基因编码的蛋白在小鼠组织中的表达。结果:重组质粒经肌肉注射免疫可产生高水平的特异性血清IgG抗体,颌下腺区注射可同时诱导高水平的唾液中sIgA和血清中IgG抗体;骨骼肌、颌下腺导管内、腺泡腔内可见阳性着色。结论:重组质粒pcDNA3. 1( ) /kgpcd经颌下腺注射可同时诱导黏膜免疫和全身免疫应答,可作为有效免疫原,这为免疫牙周炎模型定菌鼠实验提供了依据。

Immunogenicity of Porphyromonas gingivalis gene vaccine pcNDA 3.1(+)/kgp cd

Objective:To evaluate the immunogenicity of the recombinant plasmid pcDNA 3.1( )/kgp_ cd.Methods:BALB/c mice were immunized with recombinant plasmid pcDNA 3.1( )/kgp_ cd,KGP_ cd protein(control) or vector pcDNA 3.1( )(control) by quadriceps injection or targeted submandibular gland(TSG) injection. Serum IgG and salivary sIgA levels were assayed by indirect ELISA after immunization. The expression of the protein KGP_ cd in quadriceps and submandibular gland was detected by immunohistochemistry techniques.Results: Serum specific anti-KGP_ cd IgG elicited by pcDNA 3.1( )/kgp_ cd and KGP_ cd protein was significantly higher in both the quadriceps injection group and the TSG group than that in the pcDNA 3.1( ) group (P<0.01).However, the salivary sIgA level was significantly higher only in the TSG group injected with pcDNA 3.1( )/kgp_ cd (P< 0.01). Expression of the KGP_ cd was detected in the sarcoplasm, sarcolemma of muscle fibers and submandibular gland duct.Conclusion:The recombinant pcDNA 3.1( )/kgp_ cd can induce both humoral and mucosal immunal responses by TSG immunization and it may be used as a candidate gene vaccine.