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Intranasal co-delivery of IL-6 gene enhances the immunogenicity of anti-caries DNA vaccine.

Su LK, Yu F, Li ZF, Zeng C, Xu QA, Fan MW - Acta Pharmacol. Sin. (2014)

Bottom Line: 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.Marked expression of IL-6 was found in COS-7 cells transfected with pCI-IL-6.Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.

View Article: PubMed Central - PubMed

Affiliation: 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.

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.

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Related in: MedlinePlus

Levels of serum specific IgG antibody. Blood was collected from mice immunized intranasally with pCIA-P plus pCI-IL-6 or pCIA-P plus pCI vector every 4 weeks and the serum was obtained for ELISA. Mean±SEM. n=8. cP<0.01 vs pCIA-P+pCI group.
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fig3: Levels of serum specific IgG antibody. Blood was collected from mice immunized intranasally with pCIA-P plus pCI-IL-6 or pCIA-P plus pCI vector every 4 weeks and the serum was obtained for ELISA. Mean±SEM. n=8. cP<0.01 vs pCIA-P+pCI group.

Mentions: To determine the mucosal adjuvant potential of IL-6, the mice were intranasally immunized with pCIA-P plus pCI-IL-6 or pCIA-P plus pCI, and the serum was collected for ELISA detection of PAc specific IgG antibodies. The level of the serum specific IgG antibody was undetectable prior to immunization in both groups (data not shown). The mean serum specific IgG antibody levels of the pCIA-P plus pCI-IL-6 immunization group were significantly higher than those in the pCIA-P plus pCI immunization group at weeks 4 and 8 (P<0.01, Figure 3). Although the IgG levels of the IL-6 co-immunization group were higher than the control group at weeks 12 and 16, the statistical analysis showed no significant difference between the two groups (P>0.05). The highest level of serum specific IgG antibody appeared at week 8 and gradually decreased over time.


Intranasal co-delivery of IL-6 gene enhances the immunogenicity of anti-caries DNA vaccine.

Su LK, Yu F, Li ZF, Zeng C, Xu QA, Fan MW - Acta Pharmacol. Sin. (2014)

Levels of serum specific IgG antibody. Blood was collected from mice immunized intranasally with pCIA-P plus pCI-IL-6 or pCIA-P plus pCI vector every 4 weeks and the serum was obtained for ELISA. Mean±SEM. n=8. cP<0.01 vs pCIA-P+pCI group.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4814028&req=5

fig3: Levels of serum specific IgG antibody. Blood was collected from mice immunized intranasally with pCIA-P plus pCI-IL-6 or pCIA-P plus pCI vector every 4 weeks and the serum was obtained for ELISA. Mean±SEM. n=8. cP<0.01 vs pCIA-P+pCI group.
Mentions: To determine the mucosal adjuvant potential of IL-6, the mice were intranasally immunized with pCIA-P plus pCI-IL-6 or pCIA-P plus pCI, and the serum was collected for ELISA detection of PAc specific IgG antibodies. The level of the serum specific IgG antibody was undetectable prior to immunization in both groups (data not shown). The mean serum specific IgG antibody levels of the pCIA-P plus pCI-IL-6 immunization group were significantly higher than those in the pCIA-P plus pCI immunization group at weeks 4 and 8 (P<0.01, Figure 3). Although the IgG levels of the IL-6 co-immunization group were higher than the control group at weeks 12 and 16, the statistical analysis showed no significant difference between the two groups (P>0.05). The highest level of serum specific IgG antibody appeared at week 8 and gradually decreased over time.

Bottom Line: 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.Marked expression of IL-6 was found in COS-7 cells transfected with pCI-IL-6.Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.

View Article: PubMed Central - PubMed

Affiliation: 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.

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.

Show MeSH
Related in: MedlinePlus