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Superior in vitro stimulation of human CD8+ T-cells by whole virus versus split virus influenza vaccines.

Halbroth BR, Heil A, Distler E, Dass M, Wagner EM, Plachter B, Probst HC, Strand D, Hartwig UF, Karner A, Aichinger G, Kistner O, Landfester K, Herr W - PLoS ONE (2014)

Bottom Line: Our study showed that influenza whole virus vaccines of major seasonal A and B strains activated DC more efficiently than those of pandemic swine-origin H1N1 and pandemic-like avian H5N1 strains.In contrast, influenza split virus vaccines had a low ability to activate DC, regardless which strain was investigated.We conclude that vaccines against seasonal and pandemic (-like) influenza strains that aim to stimulate cross-reacting CD8+ T cells should include whole virus rather than split virus formulations.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine III - University Medical Center of Johannes Gutenberg-University, Mainz, Germany.

ABSTRACT
Pandemic and seasonal influenza viruses cause considerable morbidity and mortality in the general human population. Protection from severe disease may result from vaccines that activate antigen-presenting DC for effective stimulation of influenza-specific memory T cells. Special attention is paid to vaccine-induced CD8+ T-cell responses, because they are mainly directed against conserved internal influenza proteins thereby presumably mediating cross-protection against circulating seasonal as well as emerging pandemic virus strains. Our study showed that influenza whole virus vaccines of major seasonal A and B strains activated DC more efficiently than those of pandemic swine-origin H1N1 and pandemic-like avian H5N1 strains. In contrast, influenza split virus vaccines had a low ability to activate DC, regardless which strain was investigated. We also observed that whole virus vaccines stimulated virus-specific CD8+ memory T cells much stronger compared to split virus counterparts, whereas both vaccine formats activated CD4+ Th cell responses similarly. Moreover, our data showed that whole virus vaccine material is delivered into the cytosolic pathway of DC for effective activation of virus-specific CD8+ T cells. We conclude that vaccines against seasonal and pandemic (-like) influenza strains that aim to stimulate cross-reacting CD8+ T cells should include whole virus rather than split virus formulations.

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Split virus preparation substantially decreases the ability to activate and mature DC.(A) Immature DC were incubated for 48 h with different pandemic (-like) and seasonal influenza virus vaccines at 10 µg/mL (referring to HA content) and were subsequently analyzed by flow cytometry for expression of maturation markers on viable 7AAD-negative cells. Representative data of a single donor are shown in Fig. 1. Relative fluorescence intensity was calculated for each individual marker from MFI value of marker-specific staining divided by MFI value of the related IgG isotype control staining and was measured in 6 randomly selected donors. Summarized data are shown here as box blot diagrams. Split virus formulations were unavailable from pandemic (-like) strains A/H1N1-California and A/H5N1-Indonesia. P-values comparing the maturation effect of whole virus vaccines and related split virus vaccines on DC were calculated using two-tailed paired Wilcoxon signed-rank test (*, p<0.10; **, p<0.05). (B) After incubation of immature DC for 48 h with different pandemic (-like) and seasonal influenza virus vaccines, culture supernatants of four randomly selected donors were measured for IFN-α by ELISA, as well as for IL-6 and TNF-α by cytometric bead array. Graphs show mean concentrations (± SD) derived from experiments in all four healthy donors. For abbreviations of vaccines see Table 1. w/o, without.
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pone-0103392-g002: Split virus preparation substantially decreases the ability to activate and mature DC.(A) Immature DC were incubated for 48 h with different pandemic (-like) and seasonal influenza virus vaccines at 10 µg/mL (referring to HA content) and were subsequently analyzed by flow cytometry for expression of maturation markers on viable 7AAD-negative cells. Representative data of a single donor are shown in Fig. 1. Relative fluorescence intensity was calculated for each individual marker from MFI value of marker-specific staining divided by MFI value of the related IgG isotype control staining and was measured in 6 randomly selected donors. Summarized data are shown here as box blot diagrams. Split virus formulations were unavailable from pandemic (-like) strains A/H1N1-California and A/H5N1-Indonesia. P-values comparing the maturation effect of whole virus vaccines and related split virus vaccines on DC were calculated using two-tailed paired Wilcoxon signed-rank test (*, p<0.10; **, p<0.05). (B) After incubation of immature DC for 48 h with different pandemic (-like) and seasonal influenza virus vaccines, culture supernatants of four randomly selected donors were measured for IFN-α by ELISA, as well as for IL-6 and TNF-α by cytometric bead array. Graphs show mean concentrations (± SD) derived from experiments in all four healthy donors. For abbreviations of vaccines see Table 1. w/o, without.

Mentions: Whole virus vaccines were analyzed for the ability to induce maturation in monocyte-derived immature DC of healthy individuals. Vaccines were available from seasonal strains A/H1N1-Brisbane, A/H3N2-Uruguay, and B/Brisbane, as well as from recent pandemic-like avian and pandemic swine-origin strains A/H5N1-Indonesia, A/H5N1-Vietnam, and A/H1N1-California, respectively (Table 1). Flow cytometry data on DC showed that incubation with seasonal whole virus vaccines strongly increased the expression of CD80, CD86, HLA-A/B/C, HLA-DR, CD40, CD83, consistent with a phenotype of mature DC (Fig. 1, Fig. 2A). This up-regulation was absent or observed at much lower level if pandemic (-like) whole virus vaccines were added to DC.


Superior in vitro stimulation of human CD8+ T-cells by whole virus versus split virus influenza vaccines.

Halbroth BR, Heil A, Distler E, Dass M, Wagner EM, Plachter B, Probst HC, Strand D, Hartwig UF, Karner A, Aichinger G, Kistner O, Landfester K, Herr W - PLoS ONE (2014)

Split virus preparation substantially decreases the ability to activate and mature DC.(A) Immature DC were incubated for 48 h with different pandemic (-like) and seasonal influenza virus vaccines at 10 µg/mL (referring to HA content) and were subsequently analyzed by flow cytometry for expression of maturation markers on viable 7AAD-negative cells. Representative data of a single donor are shown in Fig. 1. Relative fluorescence intensity was calculated for each individual marker from MFI value of marker-specific staining divided by MFI value of the related IgG isotype control staining and was measured in 6 randomly selected donors. Summarized data are shown here as box blot diagrams. Split virus formulations were unavailable from pandemic (-like) strains A/H1N1-California and A/H5N1-Indonesia. P-values comparing the maturation effect of whole virus vaccines and related split virus vaccines on DC were calculated using two-tailed paired Wilcoxon signed-rank test (*, p<0.10; **, p<0.05). (B) After incubation of immature DC for 48 h with different pandemic (-like) and seasonal influenza virus vaccines, culture supernatants of four randomly selected donors were measured for IFN-α by ELISA, as well as for IL-6 and TNF-α by cytometric bead array. Graphs show mean concentrations (± SD) derived from experiments in all four healthy donors. For abbreviations of vaccines see Table 1. w/o, without.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4114834&req=5

pone-0103392-g002: Split virus preparation substantially decreases the ability to activate and mature DC.(A) Immature DC were incubated for 48 h with different pandemic (-like) and seasonal influenza virus vaccines at 10 µg/mL (referring to HA content) and were subsequently analyzed by flow cytometry for expression of maturation markers on viable 7AAD-negative cells. Representative data of a single donor are shown in Fig. 1. Relative fluorescence intensity was calculated for each individual marker from MFI value of marker-specific staining divided by MFI value of the related IgG isotype control staining and was measured in 6 randomly selected donors. Summarized data are shown here as box blot diagrams. Split virus formulations were unavailable from pandemic (-like) strains A/H1N1-California and A/H5N1-Indonesia. P-values comparing the maturation effect of whole virus vaccines and related split virus vaccines on DC were calculated using two-tailed paired Wilcoxon signed-rank test (*, p<0.10; **, p<0.05). (B) After incubation of immature DC for 48 h with different pandemic (-like) and seasonal influenza virus vaccines, culture supernatants of four randomly selected donors were measured for IFN-α by ELISA, as well as for IL-6 and TNF-α by cytometric bead array. Graphs show mean concentrations (± SD) derived from experiments in all four healthy donors. For abbreviations of vaccines see Table 1. w/o, without.
Mentions: Whole virus vaccines were analyzed for the ability to induce maturation in monocyte-derived immature DC of healthy individuals. Vaccines were available from seasonal strains A/H1N1-Brisbane, A/H3N2-Uruguay, and B/Brisbane, as well as from recent pandemic-like avian and pandemic swine-origin strains A/H5N1-Indonesia, A/H5N1-Vietnam, and A/H1N1-California, respectively (Table 1). Flow cytometry data on DC showed that incubation with seasonal whole virus vaccines strongly increased the expression of CD80, CD86, HLA-A/B/C, HLA-DR, CD40, CD83, consistent with a phenotype of mature DC (Fig. 1, Fig. 2A). This up-regulation was absent or observed at much lower level if pandemic (-like) whole virus vaccines were added to DC.

Bottom Line: Our study showed that influenza whole virus vaccines of major seasonal A and B strains activated DC more efficiently than those of pandemic swine-origin H1N1 and pandemic-like avian H5N1 strains.In contrast, influenza split virus vaccines had a low ability to activate DC, regardless which strain was investigated.We conclude that vaccines against seasonal and pandemic (-like) influenza strains that aim to stimulate cross-reacting CD8+ T cells should include whole virus rather than split virus formulations.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine III - University Medical Center of Johannes Gutenberg-University, Mainz, Germany.

ABSTRACT
Pandemic and seasonal influenza viruses cause considerable morbidity and mortality in the general human population. Protection from severe disease may result from vaccines that activate antigen-presenting DC for effective stimulation of influenza-specific memory T cells. Special attention is paid to vaccine-induced CD8+ T-cell responses, because they are mainly directed against conserved internal influenza proteins thereby presumably mediating cross-protection against circulating seasonal as well as emerging pandemic virus strains. Our study showed that influenza whole virus vaccines of major seasonal A and B strains activated DC more efficiently than those of pandemic swine-origin H1N1 and pandemic-like avian H5N1 strains. In contrast, influenza split virus vaccines had a low ability to activate DC, regardless which strain was investigated. We also observed that whole virus vaccines stimulated virus-specific CD8+ memory T cells much stronger compared to split virus counterparts, whereas both vaccine formats activated CD4+ Th cell responses similarly. Moreover, our data showed that whole virus vaccine material is delivered into the cytosolic pathway of DC for effective activation of virus-specific CD8+ T cells. We conclude that vaccines against seasonal and pandemic (-like) influenza strains that aim to stimulate cross-reacting CD8+ T cells should include whole virus rather than split virus formulations.

Show MeSH
Related in: MedlinePlus