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1.
Acta Pharmacol Sin ; 35(5): 592-8, 2014 May.
Article in English | MEDLINE | ID: mdl-24705100

ABSTRACT

AIM: To investigate the effects of co-delivering IL-6 expressing plasmid pCI-IL-6 on the immunogenicity of the anti-caries DNA vaccine pCIA-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 pCIA-P plus pCI-IL-6 on d 0 and 14. Anti-PAc IgG and secretory IgA (sIgA) were assessed by ELISA. Splenocytes from the mice were re-stimulated with the PAc protein, and IFN-γ 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 COS-7 cells transfected with pCI-IL-6. In the pCI-IL-6 co-immunized mice, the specific IgG antibodies in serum and sIgA antibodies in saliva were significantly higher than those in the control mice at weeks 4 and 8. Moreover, the secretion of IFN-γ 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 pCIA-P and pCI 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.


Subject(s)
Antibody Formation/genetics , Antibody Formation/immunology , Dental Caries/immunology , Interleukin-6/genetics , Interleukin-6/immunology , Vaccines, DNA/genetics , Vaccines, DNA/immunology , Administration, Intranasal , Animals , Antibodies, Bacterial/immunology , Antigens, Surface/immunology , Bacterial Proteins/genetics , Bacterial Proteins/immunology , COS Cells , Cell Line , Cell Proliferation , Chlorocebus aethiops , Female , Genetic Vectors/genetics , Genetic Vectors/immunology , Immunization/methods , Immunoglobulin A, Secretory/immunology , Interferon-gamma/immunology , Interleukin-4/immunology , Mice , Mice, Inbred BALB C , Plasmids/genetics , Plasmids/immunology , Rats , Rats, Wistar , Saliva/immunology , Streptococcus mutans/immunology
2.
Acta Pharmacol Sin ; 34(3): 432-40, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23334235

ABSTRACT

AIM: To investigate how co-delivery of the gene encoding C-C chemokine ligand-19 (CCL-19) affected the systemic immune responses to an anti-caries DNA vaccine pCIA-P in mice. METHODS: Plasmid encoding CCL19-GFP fusion protein (pCCL19/GFP) was constructed by inserting murine ccl19 gene into GFP-expressing vector pAcGFP1-N1. Chemotactic effect of the fusion protein on murine dendritic cells (DCs) was assessed in vitro and in vivo using transwell and flow cytometric analysis, respectively. BALB/c mice were administered anti-caries DNA vaccine pCIA-P plus pCCL19/GFP (each 100 µg, im) or pCIA-P alone. Serum level of anti-PAc IgG was assessed with ELISA. Splenocytes from the mice were stimulated with PAc protein for 48 h, and IFN-γ and IL-4 production was measured with ELISA. The presence of pCCL19/GFP in spleen and draining lymph nodes was assessed using PCR. The expression of pCCL19/GFP protein in these tissues was analyzed under microscope and with flow cytometry. RESULTS: The expression level of CCL19-GFP fusion protein was considerably increased 48 h after transfection of COS-7 cells with pCCL19/GFP plasmids. The fusion protein showed potent chemotactic activity on DCs in vitro. The level of serum PAc-specific IgG was significantly increased from 4 to 14 weeks in the mice vaccinated with pCIA-P plus pCCL19/GFP. Compared to mice vaccinated with pCIA-P alone, the splenocytes from mice vaccinated with pCIA-P plus pCCL19/GFP produced significantly higher level of IFN-γ, but IL-4 production had no significant change. Following intromuscular co-delivery, pCCL19/GFP plasmid and fusion protein were detected in the spleen and draining lymph nodes. Administration of CCL19 gene in mice markedly increased the number of mature DCs in secondary lymphoid tissues. CONCLUSION: CCL19 serves as an effective adjuvant for anti-caries DNA vaccine by inducing chemotactic migration of DCs to secondary lymphoid tissues.


Subject(s)
Chemokine CCL19/genetics , Chemotaxis/immunology , Dendritic Cells/immunology , Dental Caries/prevention & control , Lymph Nodes/immunology , Spleen/immunology , Vaccines, DNA/immunology , Animals , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , COS Cells , Chemokine CCL19/administration & dosage , Chemokine CCL19/immunology , Chemotaxis/genetics , Chlorocebus aethiops , Cytokines/immunology , Dendritic Cells/cytology , Dental Caries/immunology , Dental Caries/microbiology , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Genetic Vectors , Green Fluorescent Proteins/genetics , Injections, Intramuscular , Mice , Plasmids , Recombinant Fusion Proteins/genetics , Streptococcus mutans/genetics , Streptococcus mutans/immunology , Transfection , Vaccines, DNA/genetics
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