牙龈卟啉单胞菌Kgp DNA疫苗防治牙周炎的实验研究

Porphyromonas Gingivalis Kgp DNA Vaccine Induces Antibody Response in Experimental Periodontitis

目的构建牙龈卟啉单胞菌(P.gingivalis)Kgp DNA疫苗真核表达质粒,并对Wistar大鼠进行免疫接种,分析和评价其对牙周炎的免疫效果。方法将牙龈卟啉单胞菌ATCC33277 Kgp基因通过聚合酶链反应(PCR)扩增后连接至真核表达质粒pVAX1,构建真核表达质粒pVAX1-kgp;16只Wistar大鼠随机分为A,B两组,分别接种pVAX1-Kgp和pVAX1,绑线法构建牙周炎模型,记录各组牙周状况,免疫前后收集大鼠唾液和血清样本,酶联免疫吸附测定(ELISA)检测唾液中sIgA和外周血中IgG抗体水平。结果 A组牙周软组织炎症和牙槽骨吸收明显低于B组,且A组动物sIgA和IgG抗体较未免疫前明显增高(P<0.05),较B组显示出更高的抗体水平(P<0.05)。结论牙龈卟啉单胞菌Kgp基因疫苗可诱导牙周炎大鼠产生有效免疫应答,对实验性牙周炎起到明显保护作用。

Objective To construct Kgp gene vaccine and to evaluate its immune response in animal model with periodontitis,and to provide experimental data for the study of peri-implantits. Methods Kgp gene amplified from Poqhyromonas gingivalis strains ATCC33277 were cloned into vector pVAX1.16 rats and randomly divided into group A and group B. Grot, p A was immunized with pVAXI-Kgp,while group B with pVAX1. Cotton ligation infiltrated with Porphyromonas gingivalis was ligatured around the neck of the first maxillary molar to construct the periodontitis model. The status of periodontal tissue was ohserved. Before the first immunized and one week after the last imnmnized,slgA and lgG specific antibody were detected by EL1SA. Results After imnmnization,slgA and IgG of group A were higher than before the immunization （ P 〈 0.05 ） , while the difference in group B had no statistically significant. Furthermore after im munization, antihodies of group A showed higher titres than group B （ P 〈 0.05 ） , before immunization, these two groups showed no difference（ P 〉0.05）. Periodontitis in group B was more serious than in group A. Conclusion Kgp DNA vaccine couht induce specific immune response in experimental periodontitis, and these could be effective to protect bone loss.