Kong F, Zheng D, She P, Ni P, Zhu H, Xu H, Su Z
Department of Stomatology, the Affiliated People's Hospital of Jiangsu University, Zhenjiang, China.
Department of Immunology & Laboratory Immunology, Jiangsu University, Zhenjiang, China.
Scand J Immunol. 2015 Jun;81(6):476-82. doi: 10.1111/sji.12281.
The outer membrane protein RagB is one of the major virulence factors of Porphyromonas gingivalis (P. gingivalis). To prevent periodontitis and associated systemic diseases induced by P. gingivalis, we built B cell antigen epitope vaccine characterized by pIRES-ragB'-mGITRL to induce a protective immune responses. The B cell antigen epitope and scrambled peptide of ragB were predicted, cloned into pIRES and constructed pIRES-ragB', pIRES-scrambled epitopes and pIRES-ragB'-mGITRL. pIRES-ragB'-mGITRL was transfected into COS-7 cells. Subsequently, the 6-week-old female BALB/c mice were challenged by P. gingivalis following three time immunization by pIRES, pIRES-ragB', pIRES-scrambled epitopes and pIRES-ragB'-mGITRL. The levels of RagB-specific antibody in the serum and Tfh cells in the spleen were measured by ELISA and flow cytometry, respectively. And higher levels of RagB-specific IgG were produced in the immunized mice with pIRES-ragB'-mGITRL. Additionally, the number of Tfh cells was also expanded and lesions were diminished in pIRES-ragB'-mGITRL mice comparing with control groups. Our results clearly demonstrated that P. gingivalis B cell antigen epitope vaccine, pIRES-ragB'-mGITRL, could induce protective immune responses. Furthermore, our data also indicated that pIRES-ragB'-mGITRL was a potential therapeutic vaccine against P. gingivalis.
外膜蛋白RagB是牙龈卟啉单胞菌(P. gingivalis)的主要毒力因子之一。为预防由牙龈卟啉单胞菌引起的牙周炎及相关全身性疾病,我们构建了以pIRES-ragB'-mGITRL为特征的B细胞抗原表位疫苗,以诱导保护性免疫反应。对ragB的B细胞抗原表位和乱序肽进行了预测,克隆至pIRES中,构建了pIRES-ragB'、pIRES-乱序表位和pIRES-ragB'-mGITRL。将pIRES-ragB'-mGITRL转染至COS-7细胞。随后,6周龄雌性BALB/c小鼠在经pIRES、pIRES-ragB'、pIRES-乱序表位和pIRES-ragB'-mGITRL三次免疫后,受到牙龈卟啉单胞菌攻击。分别通过ELISA和流式细胞术检测血清中RagB特异性抗体水平和脾脏中Tfh细胞水平。用pIRES-ragB'-mGITRL免疫的小鼠产生了更高水平的RagB特异性IgG。此外,与对照组相比,pIRES-ragB'-mGITRL小鼠中Tfh细胞数量也增加,病变减轻。我们的结果清楚地表明,牙龈卟啉单胞菌B细胞抗原表位疫苗pIRES-ragB'-mGITRL可诱导保护性免疫反应。此外,我们的数据还表明,pIRES-ragB'-mGITRL是一种针对牙龈卟啉单胞菌的潜在治疗性疫苗。