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The outer membrane protein RagB is one of the major virulence factors of the periodontal pathogen  Porphyromonas gingivalis  ( P. gingivalis ). In order to induce protective immune response against  P. gingivalis  infection, an  mGITRL  gene-linked  ragB  DNA vaccine (pIRES -ragB-mGITRL  ) was constructed. Six-week-old female BALB/c mice were immunized with pIRES -ragB-mGITRL  through intramuscular injection and then challenged by subcutaneous injection in the abdomen with  P. gingivalis . RagB-specific antibody-forming cells were evaluated by an Enzyme-linked immunosorbent spot, and specific antibody was determined by enzyme-linked immunosorbent assay. In addition, the frequencies of Tfh and IFN-γ +  T cells in spleen were measured using flow cytometer, and the levels of IL-21 and IFN-γ mRNA or proteins were detected by real time RT-PCR or ELISA. The data showed that the  mGITRL -linked  ragB  DNA vaccine induced higher levels of RagB-specific IgG in serum and RagB-specific antibody-forming cells in spleen. The frequencies of Tfh and IFN-γ +  T cells were obviously expanded in mice immunized by pIRES -ragB-mGITRL  compared with other groups (pIRES  or  pIRES -ragB  ). The levels of Tfh and IFN-γ +  T cells associated cytokines were also significantly increased in pIRES- ragB-mGITRL  group. Therefore, the mice immunized with  ragB  plus  mGITRL  showed the stronger resistant to  P. gingivalis  infection and a significant reduction of the lesion size caused by  P. gingivalis  infection comparing with other groups. Taken together, our findings demonstrated that intramuscular injection of DNA vaccine  ragB  together with  mGITRL  induced protective immune response dramatically by increasing Tfh and IFN-γ +  T cells and antibody production to  P. gingivalis .

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Enhancing Specific-Antibody Production to the ragB Vaccine with GITRL That Expand Tfh, IFN-γ+ T Cells and Attenuates Porphyromonas gingivalis Infection in Mice