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Evaluation of cellular responses for a chimeric HBsAg-HCV core DNA vaccine in BALB/c mice.

Yazdanian M, Memarnejadian A, Mahdavi M, Motevalli F, Sadat SM, Vahabpour R, Khanahmad H, Soleimanjahi H, Budkowska A, Roohvand F - Adv Biomed Res (2015)

Bottom Line: The cellular immune responses (proliferation, In vivo CTL assay and IFN-γ/IL-4 ELISpot) against a strong and dominant H2-d restricted, CD8(+)-epitopic peptide (C39) (core 39-48; RRGPRLGVRA) of HCVcp were compared in immunized animals.Proper expression of the fused protein by pCHCORE in transiently transfected HEK 293T cells and in the expected size (around 50 kDa) was confirmed by western blotting.Fusion of HBsAg to HCVcp in the context of a DNA vaccine modality could augment Th1-oriented cellular and CTL responses toward a protective epitope, comparable to that of HCVcp (subunit HCV vaccine) immunization.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.

ABSTRACT

Background: Fusion of Hepatitis B virus surface antigen (HBsAg) to a DNA construct might be considered as a strategy to enhance cellular and cytotoxic T-lymphocytes (CTL) responses of a Hepatitis C Virus core protein (HCVcp)-based DNA vaccine comparable to that of adjuvanted protein (subunit) immunization.

Materials and methods: pCHCORE vector harboring coding sequence of HBsAg and HCVcp (amino acids 2-120) in tandem within the pCDNA3.1 backbone was constructed. The corresponding recombinant HCVcp was also expressed and purified in Escherichia coli. Mice were immunized either by adjuvanted HCVcp (pluronic acid + protein) or by pCHCORE vector primed/protein boosted immunization regimen. The cellular immune responses (proliferation, In vivo CTL assay and IFN-γ/IL-4 ELISpot) against a strong and dominant H2-d restricted, CD8(+)-epitopic peptide (C39) (core 39-48; RRGPRLGVRA) of HCVcp were compared in immunized animals.

Result: Proper expression of the fused protein by pCHCORE in transiently transfected HEK 293T cells and in the expected size (around 50 kDa) was confirmed by western blotting. The immunization results indicated that the pCHCORE shifted the immune responses pathway to Th1 by enhancing the IFN-γ cytokine level much higher than protein immunization while the proliferative and CTL responses were comparable (or slightly in favor of DNA immunization).

Conclusion: Fusion of HBsAg to HCVcp in the context of a DNA vaccine modality could augment Th1-oriented cellular and CTL responses toward a protective epitope, comparable to that of HCVcp (subunit HCV vaccine) immunization.

No MeSH data available.


Related in: MedlinePlus

Enzyme-linked-immunospot assays. Mouse Elispot kit (Mabtech, Sweden) was used for detection of class I-binding C39 peptide specific IFN-γ and IL-4-releasing T cells in splenocytes of immunized mice. Results are shown as the numbers of spot-forming-cells per 106 splenocytes
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Figure 4: Enzyme-linked-immunospot assays. Mouse Elispot kit (Mabtech, Sweden) was used for detection of class I-binding C39 peptide specific IFN-γ and IL-4-releasing T cells in splenocytes of immunized mice. Results are shown as the numbers of spot-forming-cells per 106 splenocytes

Mentions: In accordance with the results of proliferation and CTL assays, data of ex vivo ELISpot assay also indicated a significant difference in the number of IFN-γ and IL-4-secreting cells in favor of pCHCORE and HCVcp immunized mice groups compared with the control groups [Figure 4]. Moreover, as shown in Figure 4, the group immunized with pCHCORE elicited significantly more spots of IFN-γ than HCVcp vaccinated groups. In contrast, the immunized group by HCVcp shows the IL-4 secreting responses significantly higher than the vaccinated group by pCHCORE. These results clearly indicated an elevated Th1-oriented response for our DNA vaccination strategy compared with that of protein immunization.


Evaluation of cellular responses for a chimeric HBsAg-HCV core DNA vaccine in BALB/c mice.

Yazdanian M, Memarnejadian A, Mahdavi M, Motevalli F, Sadat SM, Vahabpour R, Khanahmad H, Soleimanjahi H, Budkowska A, Roohvand F - Adv Biomed Res (2015)

Enzyme-linked-immunospot assays. Mouse Elispot kit (Mabtech, Sweden) was used for detection of class I-binding C39 peptide specific IFN-γ and IL-4-releasing T cells in splenocytes of immunized mice. Results are shown as the numbers of spot-forming-cells per 106 splenocytes
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300588&req=5

Figure 4: Enzyme-linked-immunospot assays. Mouse Elispot kit (Mabtech, Sweden) was used for detection of class I-binding C39 peptide specific IFN-γ and IL-4-releasing T cells in splenocytes of immunized mice. Results are shown as the numbers of spot-forming-cells per 106 splenocytes
Mentions: In accordance with the results of proliferation and CTL assays, data of ex vivo ELISpot assay also indicated a significant difference in the number of IFN-γ and IL-4-secreting cells in favor of pCHCORE and HCVcp immunized mice groups compared with the control groups [Figure 4]. Moreover, as shown in Figure 4, the group immunized with pCHCORE elicited significantly more spots of IFN-γ than HCVcp vaccinated groups. In contrast, the immunized group by HCVcp shows the IL-4 secreting responses significantly higher than the vaccinated group by pCHCORE. These results clearly indicated an elevated Th1-oriented response for our DNA vaccination strategy compared with that of protein immunization.

Bottom Line: The cellular immune responses (proliferation, In vivo CTL assay and IFN-γ/IL-4 ELISpot) against a strong and dominant H2-d restricted, CD8(+)-epitopic peptide (C39) (core 39-48; RRGPRLGVRA) of HCVcp were compared in immunized animals.Proper expression of the fused protein by pCHCORE in transiently transfected HEK 293T cells and in the expected size (around 50 kDa) was confirmed by western blotting.Fusion of HBsAg to HCVcp in the context of a DNA vaccine modality could augment Th1-oriented cellular and CTL responses toward a protective epitope, comparable to that of HCVcp (subunit HCV vaccine) immunization.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.

ABSTRACT

Background: Fusion of Hepatitis B virus surface antigen (HBsAg) to a DNA construct might be considered as a strategy to enhance cellular and cytotoxic T-lymphocytes (CTL) responses of a Hepatitis C Virus core protein (HCVcp)-based DNA vaccine comparable to that of adjuvanted protein (subunit) immunization.

Materials and methods: pCHCORE vector harboring coding sequence of HBsAg and HCVcp (amino acids 2-120) in tandem within the pCDNA3.1 backbone was constructed. The corresponding recombinant HCVcp was also expressed and purified in Escherichia coli. Mice were immunized either by adjuvanted HCVcp (pluronic acid + protein) or by pCHCORE vector primed/protein boosted immunization regimen. The cellular immune responses (proliferation, In vivo CTL assay and IFN-γ/IL-4 ELISpot) against a strong and dominant H2-d restricted, CD8(+)-epitopic peptide (C39) (core 39-48; RRGPRLGVRA) of HCVcp were compared in immunized animals.

Result: Proper expression of the fused protein by pCHCORE in transiently transfected HEK 293T cells and in the expected size (around 50 kDa) was confirmed by western blotting. The immunization results indicated that the pCHCORE shifted the immune responses pathway to Th1 by enhancing the IFN-γ cytokine level much higher than protein immunization while the proliferative and CTL responses were comparable (or slightly in favor of DNA immunization).

Conclusion: Fusion of HBsAg to HCVcp in the context of a DNA vaccine modality could augment Th1-oriented cellular and CTL responses toward a protective epitope, comparable to that of HCVcp (subunit HCV vaccine) immunization.

No MeSH data available.


Related in: MedlinePlus