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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

Restriction pattern profiles of the digested plasmid. The pCI-IL-6 plasmid was enzymatically digested, and the digests were subjected to agarose gel electrophoresis. M lane, DNA Ladder; Lane 1, undigested plasmid pCI; Lane 2, EcoR I digested plasmid; Lane 3, EcoR I and Sal I digested plasmid; Lane 4, PCR product of target gene.
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fig1: Restriction pattern profiles of the digested plasmid. The pCI-IL-6 plasmid was enzymatically digested, and the digests were subjected to agarose gel electrophoresis. M lane, DNA Ladder; Lane 1, undigested plasmid pCI; Lane 2, EcoR I digested plasmid; Lane 3, EcoR I and Sal I digested plasmid; Lane 4, PCR product of target gene.

Mentions: The pCI-IL-6 plasmid was constructed as described above and confirmed by PCR, enzyme digestion and sequence analysis. Electrophoresis after digestion showed that the il-6 gene was properly inserted into the vector plasmid pCI (Figure 1). Sequence analysis showed that the sequence of the cloned IL-6 gene was identical to the sequence in Genbank (Nucleotide ID: NM_031168.1). Plasmid expression was confirmed by Western blot, which showed that the expression of IL-6 in the pCI-IL-6 transfected group was higher than that in the pCI transfected group (Figure 2).


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)

Restriction pattern profiles of the digested plasmid. The pCI-IL-6 plasmid was enzymatically digested, and the digests were subjected to agarose gel electrophoresis. M lane, DNA Ladder; Lane 1, undigested plasmid pCI; Lane 2, EcoR I digested plasmid; Lane 3, EcoR I and Sal I digested plasmid; Lane 4, PCR product of target gene.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4814028&req=5

fig1: Restriction pattern profiles of the digested plasmid. The pCI-IL-6 plasmid was enzymatically digested, and the digests were subjected to agarose gel electrophoresis. M lane, DNA Ladder; Lane 1, undigested plasmid pCI; Lane 2, EcoR I digested plasmid; Lane 3, EcoR I and Sal I digested plasmid; Lane 4, PCR product of target gene.
Mentions: The pCI-IL-6 plasmid was constructed as described above and confirmed by PCR, enzyme digestion and sequence analysis. Electrophoresis after digestion showed that the il-6 gene was properly inserted into the vector plasmid pCI (Figure 1). Sequence analysis showed that the sequence of the cloned IL-6 gene was identical to the sequence in Genbank (Nucleotide ID: NM_031168.1). Plasmid expression was confirmed by Western blot, which showed that the expression of IL-6 in the pCI-IL-6 transfected group was higher than that in the pCI transfected group (Figure 2).

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