ntranasal co-delivery of IL-6 gene enhances the mmunogenicity of anti-caries DNA vaccine  

作　　者：Ling-kai SU Fei YU Zhao-fei LI Chang ZENG Qing-an xu Ming-wen FAN 

机构地区：[1]The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine of Ministry of Education (KLOBM), School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China [2]Center of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

出　　处：《Acta Pharmacologica Sinica》2014年第5期592-598,共7页中国药理学报（英文版）

基　　金：This work was supported by National Natural Science Foundation of China （No 81000435 and 81271129）.Ming-wen FAN and Qing-an XU designed the study and revised the manuscript; Ling-kai SU, Fei YU, Zhao-fei LI, and Chang ZENG performed the experiments; and Ling-kai SU wrote the manuscript.

摘　　要：Aim： To investigate the effects of co-delivering 11_-6 expressing plasmid pCI-IL-6 on the immunogenicity of the anti-caries DNA vaccine pClA-P, which encodes the surface protein antigen PAc of Streptococcus mutans. Methods： Plasmid pCI-IL-6 was constructed by inserting the murine IL-6 gene into the pCI vector. Expression of IL-6 in vitro was assessed using Western blot analysis. BALB/c mice were intranasally co-immunized with pClA-P plus pCI-IL-6 on d 0 and 14. Anti-PAc IgG and secretory IgA （slgA） were assessed by ELISA. Splenocytes from the mice were re-stimulated with the PAc protein, and IFN-y and IL-4 production was measured using ELISA. Splenocyte proliferation was analyzed with flow cytometry. Rats were similarly immunized, and dental caries scores were determined using the Keyes method. Results： Marked expression of IL-6 was found in C0S-7 cells transfected with pCI-IL-6. In the pCI-IL-6 co-immunized mice, the specific IgG antibodies in serum and slgA antibodies in saliva were significantly higher than those in the control mice at weeks 4 and 8. Moreover, the secretion of IFN-y from splenocytes in response to re-stimulation with PAc protein was significantly higher in the pCI-IL-6 co-immunized mice than that in the control mice, whereas the secretion of IL-4 had no significant difference. The proliferation of splenocytes from the pCI-IL-6 co-immunized mice was significantly higher than that from the mice immunized with pClA-P and pCl vector. In the rat caries model, the pCI-IL-6 co-immunization rats displayed lower caries scores than the control rats. Conclusion： Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.Aim： To investigate the effects of co-delivering 11_-6 expressing plasmid pCI-IL-6 on the immunogenicity of the anti-caries DNA vaccine pClA-P, which encodes the surface protein antigen PAc of Streptococcus mutans. Methods： Plasmid pCI-IL-6 was constructed by inserting the murine IL-6 gene into the pCI vector. Expression of IL-6 in vitro was assessed using Western blot analysis. BALB/c mice were intranasally co-immunized with pClA-P plus pCI-IL-6 on d 0 and 14. Anti-PAc IgG and secretory IgA （slgA） were assessed by ELISA. Splenocytes from the mice were re-stimulated with the PAc protein, and IFN-y and IL-4 production was measured using ELISA. Splenocyte proliferation was analyzed with flow cytometry. Rats were similarly immunized, and dental caries scores were determined using the Keyes method. Results： Marked expression of IL-6 was found in C0S-7 cells transfected with pCI-IL-6. In the pCI-IL-6 co-immunized mice, the specific IgG antibodies in serum and slgA antibodies in saliva were significantly higher than those in the control mice at weeks 4 and 8. Moreover, the secretion of IFN-y from splenocytes in response to re-stimulation with PAc protein was significantly higher in the pCI-IL-6 co-immunized mice than that in the control mice, whereas the secretion of IL-4 had no significant difference. The proliferation of splenocytes from the pCI-IL-6 co-immunized mice was significantly higher than that from the mice immunized with pClA-P and pCl vector. In the rat caries model, the pCI-IL-6 co-immunization rats displayed lower caries scores than the control rats. Conclusion： Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.

关 键 词：DNA vaccine vaccine adjuvant dental caries Streptococcus mutans IL-6 CYTOKINE IMMUNOGENICITY STOMATOLOGY 

分 类 号：S858.28[农业科学—临床兽医学] S942.5[农业科学—兽医学]