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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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CD8+ T-cell reactivity to whole virus is superior compared to split virus preparations.CD4+ and CD8+ T cells purified from PBMC of healthy individuals were screened for IFN-γ ELISpot reactivity to autologous DC pre-loaded with 10 µg/mL of influenza whole virus and related split virus vaccine formulations. DC also received maturation cytokines during vaccine pulsing. (A) Representative data obtained from donor HD20 with 1×105 CD4+ T cells (grey columns) or 1×105 CD8+ T cells (black columns) plated per well are shown. (B, C) Reactivity to influenza whole virus and related split virus vaccines were measured in 10 randomly selected healthy individuals as described in (A). Box plot diagrams include IFN-γ ELISpot data from purified CD4+ (B) and CD8+ (C) T cells. Effective SFC were determined by subtraction of background spot numbers (w/o vaccine) from spot numbers induced by each individual vaccine. P-values were calculated by two-tailed paired-sample Wilcoxon signed-rank test.
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pone-0103392-g003: CD8+ T-cell reactivity to whole virus is superior compared to split virus preparations.CD4+ and CD8+ T cells purified from PBMC of healthy individuals were screened for IFN-γ ELISpot reactivity to autologous DC pre-loaded with 10 µg/mL of influenza whole virus and related split virus vaccine formulations. DC also received maturation cytokines during vaccine pulsing. (A) Representative data obtained from donor HD20 with 1×105 CD4+ T cells (grey columns) or 1×105 CD8+ T cells (black columns) plated per well are shown. (B, C) Reactivity to influenza whole virus and related split virus vaccines were measured in 10 randomly selected healthy individuals as described in (A). Box plot diagrams include IFN-γ ELISpot data from purified CD4+ (B) and CD8+ (C) T cells. Effective SFC were determined by subtraction of background spot numbers (w/o vaccine) from spot numbers induced by each individual vaccine. P-values were calculated by two-tailed paired-sample Wilcoxon signed-rank test.

Mentions: Vaccine-loaded matured DC were subsequently used as APC to stimulate autologous CD4+ and CD8+ T cells of 10 healthy individuals in IFN-γ ELISpot assay. As shown in detail for a single representative donor, IFN-γ secreting CD4+ and CD8+ T cells were observed upon stimulation with every influenza whole virus vaccine preparation tested (Fig. 3A). Data summarized from the entire study cohort demonstrated that the frequencies of whole virus vaccine reactive CD4+ precursors were highest for seasonal strains A/H3N2-Uruguay and B/Brisbane, and were slightly lower for seasonal strain A/H1N1-Brisbane and all pandemic (-like) strains (Fig. 3B). Median numbers (and ranges) per 105 CD4+ T cells were 105 (30–217), 110 (23–324), 112 (23–300), 87 (23–204), 135 (13–245), and 126 (5–255) for swine-origin A/H1N1-California, avian A/H5N1-Indonesia, avian A/H5N1-Vietnam, A/H1N1-Brisbane, A/H3N2-Uruguay, and B/Brisbane, respectively. In contrast, frequencies of whole virus vaccine reactive CD8+ T cells were approximately 2- to 10-fold lower than that of CD4+ counterparts, as determined in the same donor cohort (Fig. 3C). Highest immunogenicity for CD8+ T cells was observed with seasonal vaccines A/H1N1-Brisbane and A/H3N2-Uruguay, whereas CD8+ responses to seasonal vaccine B/Brisbane and all pandemic (-like) vaccines were of lower magnitude. Median values (and ranges) per 1×105 CD8+ T cells were 10 (5–54), 24 (6–173), 30 (7–173), 50 (13–211), 40 (15–223), and 18 (2–170) for A/H1N1-California, A/H5N1-Indonesia, A/H5N1-Vietnam, A/H1N1-Brisbane, A/H3N2-Uruguay, and B/Brisbane, respectively.


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)

CD8+ T-cell reactivity to whole virus is superior compared to split virus preparations.CD4+ and CD8+ T cells purified from PBMC of healthy individuals were screened for IFN-γ ELISpot reactivity to autologous DC pre-loaded with 10 µg/mL of influenza whole virus and related split virus vaccine formulations. DC also received maturation cytokines during vaccine pulsing. (A) Representative data obtained from donor HD20 with 1×105 CD4+ T cells (grey columns) or 1×105 CD8+ T cells (black columns) plated per well are shown. (B, C) Reactivity to influenza whole virus and related split virus vaccines were measured in 10 randomly selected healthy individuals as described in (A). Box plot diagrams include IFN-γ ELISpot data from purified CD4+ (B) and CD8+ (C) T cells. Effective SFC were determined by subtraction of background spot numbers (w/o vaccine) from spot numbers induced by each individual vaccine. P-values were calculated by two-tailed paired-sample Wilcoxon signed-rank test.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4114834&req=5

pone-0103392-g003: CD8+ T-cell reactivity to whole virus is superior compared to split virus preparations.CD4+ and CD8+ T cells purified from PBMC of healthy individuals were screened for IFN-γ ELISpot reactivity to autologous DC pre-loaded with 10 µg/mL of influenza whole virus and related split virus vaccine formulations. DC also received maturation cytokines during vaccine pulsing. (A) Representative data obtained from donor HD20 with 1×105 CD4+ T cells (grey columns) or 1×105 CD8+ T cells (black columns) plated per well are shown. (B, C) Reactivity to influenza whole virus and related split virus vaccines were measured in 10 randomly selected healthy individuals as described in (A). Box plot diagrams include IFN-γ ELISpot data from purified CD4+ (B) and CD8+ (C) T cells. Effective SFC were determined by subtraction of background spot numbers (w/o vaccine) from spot numbers induced by each individual vaccine. P-values were calculated by two-tailed paired-sample Wilcoxon signed-rank test.
Mentions: Vaccine-loaded matured DC were subsequently used as APC to stimulate autologous CD4+ and CD8+ T cells of 10 healthy individuals in IFN-γ ELISpot assay. As shown in detail for a single representative donor, IFN-γ secreting CD4+ and CD8+ T cells were observed upon stimulation with every influenza whole virus vaccine preparation tested (Fig. 3A). Data summarized from the entire study cohort demonstrated that the frequencies of whole virus vaccine reactive CD4+ precursors were highest for seasonal strains A/H3N2-Uruguay and B/Brisbane, and were slightly lower for seasonal strain A/H1N1-Brisbane and all pandemic (-like) strains (Fig. 3B). Median numbers (and ranges) per 105 CD4+ T cells were 105 (30–217), 110 (23–324), 112 (23–300), 87 (23–204), 135 (13–245), and 126 (5–255) for swine-origin A/H1N1-California, avian A/H5N1-Indonesia, avian A/H5N1-Vietnam, A/H1N1-Brisbane, A/H3N2-Uruguay, and B/Brisbane, respectively. In contrast, frequencies of whole virus vaccine reactive CD8+ T cells were approximately 2- to 10-fold lower than that of CD4+ counterparts, as determined in the same donor cohort (Fig. 3C). Highest immunogenicity for CD8+ T cells was observed with seasonal vaccines A/H1N1-Brisbane and A/H3N2-Uruguay, whereas CD8+ responses to seasonal vaccine B/Brisbane and all pandemic (-like) vaccines were of lower magnitude. Median values (and ranges) per 1×105 CD8+ T cells were 10 (5–54), 24 (6–173), 30 (7–173), 50 (13–211), 40 (15–223), and 18 (2–170) for A/H1N1-California, A/H5N1-Indonesia, A/H5N1-Vietnam, A/H1N1-Brisbane, A/H3N2-Uruguay, and B/Brisbane, respectively.

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