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Dendritic cell-mediated, DNA-based vaccination against hepatitis C induces the multi-epitope-specific response of humanized, HLA transgenic mice.

Mishra S, Lavelle BJ, Desrosiers J, Ardito MT, Terry F, Martin WD, De Groot AS, Gregory SH - PLoS ONE (2014)

Bottom Line: To date, therapeutic vaccines have demonstrated only limited success.Peptide-specific IFN-γ production by splenic T cells obtained at 5 weeks post-immunization was quantified by ELISpot assay; additionally, the production of IL-4, IL-10 and TNF-α were quantified by cytokine bead array.A multi-epitope-based HCV vaccine that targets DCs offers an effective approach to inducing a broad immune response and viral clearance in chronic, HCV-infected patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, Rhode Island, United States of America.

ABSTRACT
Hepatitis C virus (HCV) is the etiologic agent of chronic liver disease, hepatitis C. Spontaneous resolution of viral infection is associated with vigorous HLA class I- and class II-restricted T cell responses to multiple viral epitopes. Unfortunately, only 20% of patients clear infection spontaneously, most develop chronic disease and require therapy. The response to chemotherapy varies, however; therapeutic vaccination offers an additional treatment strategy. To date, therapeutic vaccines have demonstrated only limited success. Vector-mediated vaccination with multi-epitope-expressing DNA constructs alone or in combination with chemotherapy offers an additional treatment approach. Gene sequences encoding validated HLA-A2- and HLA-DRB1-restricted epitopes were synthesized and cloned into an expression vector. Dendritic cells (DCs) derived from humanized, HLA-A2/DRB1 transgenic (donor) mice were transfected with these multi-epitope-expressing DNA constructs. Recipient HLA-A2/DRB1 mice were vaccinated s.c. with transfected DCs; control mice received non-transfected DCs. Peptide-specific IFN-γ production by splenic T cells obtained at 5 weeks post-immunization was quantified by ELISpot assay; additionally, the production of IL-4, IL-10 and TNF-α were quantified by cytokine bead array. Splenocytes derived from vaccinated HLA-A2/DRB1 transgenic mice exhibited peptide-specific cytokine production to the vast majority of the vaccine-encoded HLA class I- and class II-restricted T cell epitopes. A multi-epitope-based HCV vaccine that targets DCs offers an effective approach to inducing a broad immune response and viral clearance in chronic, HCV-infected patients.

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Related in: MedlinePlus

Cytokine bead array.The spleens were dissected and pooled on day 35 from groups of four mice immunized with vaccine construct-transfected or non-transfected DCs. Single cell suspensions were transferred to quadruplicate wells and cultured 40 hours in the presence of the peptide indicated. The data, obtained in a single experiment representative of two experiments, are the means ± SD pg/ml cytokine produced by splenocytes obtained from mice immunized with transfected DCs minus that produced by splenocytes derived from control mice immunized with non-transfected DCs. Concentrations of IFN-γ, TNF-α, IL-4 and IL-10 (where indicated by *) are significantly greater than that produced by splenocytes derived from control animals (P<0.05; Student’s t test or Mann-Whitney Rank Sums test).
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pone-0104606-g005: Cytokine bead array.The spleens were dissected and pooled on day 35 from groups of four mice immunized with vaccine construct-transfected or non-transfected DCs. Single cell suspensions were transferred to quadruplicate wells and cultured 40 hours in the presence of the peptide indicated. The data, obtained in a single experiment representative of two experiments, are the means ± SD pg/ml cytokine produced by splenocytes obtained from mice immunized with transfected DCs minus that produced by splenocytes derived from control mice immunized with non-transfected DCs. Concentrations of IFN-γ, TNF-α, IL-4 and IL-10 (where indicated by *) are significantly greater than that produced by splenocytes derived from control animals (P<0.05; Student’s t test or Mann-Whitney Rank Sums test).

Mentions: Vaccination favored Th1-type cytokine production by splenocytes derived from vaccinated mice compared to mice inoculated with non-transfected DCs. Though the response to individual HLA-A2- and -DRB1-restricted peptides varied, the overall production of pro-inflammatory cytokines (TNF-α and IFN-γ) was elevated significantly relative to the production of IL-4 and IL-10 (Figure 5), Indeed, IL-4 production was barely detectable while IL-10 production was often not increased relative to the control.


Dendritic cell-mediated, DNA-based vaccination against hepatitis C induces the multi-epitope-specific response of humanized, HLA transgenic mice.

Mishra S, Lavelle BJ, Desrosiers J, Ardito MT, Terry F, Martin WD, De Groot AS, Gregory SH - PLoS ONE (2014)

Cytokine bead array.The spleens were dissected and pooled on day 35 from groups of four mice immunized with vaccine construct-transfected or non-transfected DCs. Single cell suspensions were transferred to quadruplicate wells and cultured 40 hours in the presence of the peptide indicated. The data, obtained in a single experiment representative of two experiments, are the means ± SD pg/ml cytokine produced by splenocytes obtained from mice immunized with transfected DCs minus that produced by splenocytes derived from control mice immunized with non-transfected DCs. Concentrations of IFN-γ, TNF-α, IL-4 and IL-10 (where indicated by *) are significantly greater than that produced by splenocytes derived from control animals (P<0.05; Student’s t test or Mann-Whitney Rank Sums test).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128787&req=5

pone-0104606-g005: Cytokine bead array.The spleens were dissected and pooled on day 35 from groups of four mice immunized with vaccine construct-transfected or non-transfected DCs. Single cell suspensions were transferred to quadruplicate wells and cultured 40 hours in the presence of the peptide indicated. The data, obtained in a single experiment representative of two experiments, are the means ± SD pg/ml cytokine produced by splenocytes obtained from mice immunized with transfected DCs minus that produced by splenocytes derived from control mice immunized with non-transfected DCs. Concentrations of IFN-γ, TNF-α, IL-4 and IL-10 (where indicated by *) are significantly greater than that produced by splenocytes derived from control animals (P<0.05; Student’s t test or Mann-Whitney Rank Sums test).
Mentions: Vaccination favored Th1-type cytokine production by splenocytes derived from vaccinated mice compared to mice inoculated with non-transfected DCs. Though the response to individual HLA-A2- and -DRB1-restricted peptides varied, the overall production of pro-inflammatory cytokines (TNF-α and IFN-γ) was elevated significantly relative to the production of IL-4 and IL-10 (Figure 5), Indeed, IL-4 production was barely detectable while IL-10 production was often not increased relative to the control.

Bottom Line: To date, therapeutic vaccines have demonstrated only limited success.Peptide-specific IFN-γ production by splenic T cells obtained at 5 weeks post-immunization was quantified by ELISpot assay; additionally, the production of IL-4, IL-10 and TNF-α were quantified by cytokine bead array.A multi-epitope-based HCV vaccine that targets DCs offers an effective approach to inducing a broad immune response and viral clearance in chronic, HCV-infected patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, Rhode Island, United States of America.

ABSTRACT
Hepatitis C virus (HCV) is the etiologic agent of chronic liver disease, hepatitis C. Spontaneous resolution of viral infection is associated with vigorous HLA class I- and class II-restricted T cell responses to multiple viral epitopes. Unfortunately, only 20% of patients clear infection spontaneously, most develop chronic disease and require therapy. The response to chemotherapy varies, however; therapeutic vaccination offers an additional treatment strategy. To date, therapeutic vaccines have demonstrated only limited success. Vector-mediated vaccination with multi-epitope-expressing DNA constructs alone or in combination with chemotherapy offers an additional treatment approach. Gene sequences encoding validated HLA-A2- and HLA-DRB1-restricted epitopes were synthesized and cloned into an expression vector. Dendritic cells (DCs) derived from humanized, HLA-A2/DRB1 transgenic (donor) mice were transfected with these multi-epitope-expressing DNA constructs. Recipient HLA-A2/DRB1 mice were vaccinated s.c. with transfected DCs; control mice received non-transfected DCs. Peptide-specific IFN-γ production by splenic T cells obtained at 5 weeks post-immunization was quantified by ELISpot assay; additionally, the production of IL-4, IL-10 and TNF-α were quantified by cytokine bead array. Splenocytes derived from vaccinated HLA-A2/DRB1 transgenic mice exhibited peptide-specific cytokine production to the vast majority of the vaccine-encoded HLA class I- and class II-restricted T cell epitopes. A multi-epitope-based HCV vaccine that targets DCs offers an effective approach to inducing a broad immune response and viral clearance in chronic, HCV-infected patients.

Show MeSH
Related in: MedlinePlus