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Vaccination with Ad5 vectors expands Ad5-specific CD8 T cells without altering memory phenotype or functionality.

Hutnick NA, Carnathan DG, Dubey SA, Cox KS, Kierstead L, Makadonas G, Ratcliffe SJ, Lasaro MO, Robertson MN, Casimiro DR, Ertl HC, Betts MR - PLoS ONE (2010)

Bottom Line: Adenoviral (Ad) vaccine vectors represent both a vehicle to present a novel antigen to the immune system as well as restimulation of immune responses against the Ad vector itself.Peripheral blood Ad5-specific CD8(+) T-cell numbers expanded after Ad5-HIV vaccination in all subjects, but differential expansion kinetics were noted in some baseline Ad5-neutralizing antibody (Ad5 nAb) seronegative subjects compared to baseline Ad5 nAb seropositive subjects.These data indicate that repeat Ad5-vector administration in humans expands Ad5-specific CD8(+) T-cells without overtly affecting their functional capacity or phenotypic properties.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Center for AIDS Research, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America.

ABSTRACT

Background: Adenoviral (Ad) vaccine vectors represent both a vehicle to present a novel antigen to the immune system as well as restimulation of immune responses against the Ad vector itself. To what degree Ad-specific CD8(+) T cells are restimulated by Ad vector vaccination is unclear, although such knowledge would be important as vector-specific CD8(+) T cell expansion could potentially further limit Ad vaccine efficacy beyond Ad-specific neutralizing antibody alone.

Methodology/principal findings: Here we addressed this issue by measuring human Adenovirus serotype 5 (Ad5)-specific CD8(+) T cells in recipients of the Merck Ad5 HIV-1 vaccine vector before, during, and after vaccination by multicolor flow cytometry. Ad5-specific CD8(+) T-cells were detectable in 95% of subjects prior to vaccination, and displayed primarily an effector-type functional profile and phenotype. Peripheral blood Ad5-specific CD8(+) T-cell numbers expanded after Ad5-HIV vaccination in all subjects, but differential expansion kinetics were noted in some baseline Ad5-neutralizing antibody (Ad5 nAb) seronegative subjects compared to baseline Ad5 nAb seropositive subjects. However, in neither group did vaccination alter polyfunctionality, mucosal targeting marker expression, or memory phenotype of Ad5-specific CD8(+) T-cells.

Conclusions: These data indicate that repeat Ad5-vector administration in humans expands Ad5-specific CD8(+) T-cells without overtly affecting their functional capacity or phenotypic properties. This is a secondary analysis of samples collected during the 016 trial. Results of the Merck 016 trial safety and immunogenicity have been previously published in the journal of clinical infectious diseases [1].

Trial registration: ClinicalTrials.gov NCT00849680[http://www.clinicaltrials.gov/show/NCT00849680].

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Related in: MedlinePlus

Ad specific T-cell functionality following vaccination.Ten seronegative (Ad5 nAb titer ≤18, five weeks 0–4, five weeks 0–78, grey circles) and five seropositive subjects (Ad5 nAb titer >18, white circles) received Merck Ad5 gag/pol/nef as described in Methods. Black lines recommend the mean. Grey asterics represent a significant increase from baseline in seronegative subjects and black asterics represent a significant difference from baseline in seropositive subjects. Black bars represent a significant difference between the serogroups at that time point A) Percent of the total Ad-specific response producing each cytokine. The total Ad-specific CD8+ response was computed by summing cells making at least IL-2, IFN-γ, MIP1α, or TNFα as measured by flow cytometry. The percentage of the total Ad-specific response was then computed for each cytokine. The percentage of the total response consisting of TNFα was significantly higher in seropositive subjects (p<0.0005) at baseline. B) Percentage of IL-2+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seronegative subjects at week 42 (p<0.01) and seropositive subjects at week 4(p = 0.015). C) Percentgae of TNFα+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seropositive subjects at week 52 (p<0.01) and 78 (p<0.02) and serpositive subjects at week 4 (p<0.01). There was a significant difference at baseline in the percentage of TNFα+ CD8+ T-cells between serogroups (p<0.05). D) Percentage of IFN-γ+ Ad-specific CD8+ T-cells. The percentage of IFN-γ+ CD8+T-cells was significantly increased above baseline in seronegative subjects at weeks 42 (p<0.004), 52 (p<0.003), and 78 (0.009) and in seropositives at weeks (4 (p<0.0001), 8 (p<0.04), 30 (p<0.01) and 42 (p<0.04). There was a significant difference in the percentage of CD8+IFN-γ+ CD8+ T-cells at week 78 between the serogroups (p<0.05). E) Percentage of MIP1α+ Ad-specific CD8+ T-cells. Seronegative subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells above baseline at weeks 26 (p<0.03), 42 (p<0.001), 52 (p<0.05) and 78 (p<0.004). Seropositive subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells at week 4 (p<0.0005) compared with baseline There was a significant difference in the percentage of MIP1α+CD8+ T-cells between the serogroups at weeks 30 (p<0.04), 42 (p<0.012), 52 (p<0.005), and 78 (p<0.001). F) Percentage of perforin+ Ad-specific CD8+ T-cells. The percentage of perforin+CD8+ T-cells was significantly increased above baseline at weeks 26 (p<0.02), 42 (p<0.001) 52 (p<0.05) and 78 (p<0.004) in seronegatives and week 4 (p<0.0005) in seropositive subject. There was a significant difference in the percentage of Perforin CD8+ T-cells at weeks 42 (p<0.04), 52 (p<0.02) and 78 (p<0.006).
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pone-0014385-g004: Ad specific T-cell functionality following vaccination.Ten seronegative (Ad5 nAb titer ≤18, five weeks 0–4, five weeks 0–78, grey circles) and five seropositive subjects (Ad5 nAb titer >18, white circles) received Merck Ad5 gag/pol/nef as described in Methods. Black lines recommend the mean. Grey asterics represent a significant increase from baseline in seronegative subjects and black asterics represent a significant difference from baseline in seropositive subjects. Black bars represent a significant difference between the serogroups at that time point A) Percent of the total Ad-specific response producing each cytokine. The total Ad-specific CD8+ response was computed by summing cells making at least IL-2, IFN-γ, MIP1α, or TNFα as measured by flow cytometry. The percentage of the total Ad-specific response was then computed for each cytokine. The percentage of the total response consisting of TNFα was significantly higher in seropositive subjects (p<0.0005) at baseline. B) Percentage of IL-2+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seronegative subjects at week 42 (p<0.01) and seropositive subjects at week 4(p = 0.015). C) Percentgae of TNFα+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seropositive subjects at week 52 (p<0.01) and 78 (p<0.02) and serpositive subjects at week 4 (p<0.01). There was a significant difference at baseline in the percentage of TNFα+ CD8+ T-cells between serogroups (p<0.05). D) Percentage of IFN-γ+ Ad-specific CD8+ T-cells. The percentage of IFN-γ+ CD8+T-cells was significantly increased above baseline in seronegative subjects at weeks 42 (p<0.004), 52 (p<0.003), and 78 (0.009) and in seropositives at weeks (4 (p<0.0001), 8 (p<0.04), 30 (p<0.01) and 42 (p<0.04). There was a significant difference in the percentage of CD8+IFN-γ+ CD8+ T-cells at week 78 between the serogroups (p<0.05). E) Percentage of MIP1α+ Ad-specific CD8+ T-cells. Seronegative subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells above baseline at weeks 26 (p<0.03), 42 (p<0.001), 52 (p<0.05) and 78 (p<0.004). Seropositive subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells at week 4 (p<0.0005) compared with baseline There was a significant difference in the percentage of MIP1α+CD8+ T-cells between the serogroups at weeks 30 (p<0.04), 42 (p<0.012), 52 (p<0.005), and 78 (p<0.001). F) Percentage of perforin+ Ad-specific CD8+ T-cells. The percentage of perforin+CD8+ T-cells was significantly increased above baseline at weeks 26 (p<0.02), 42 (p<0.001) 52 (p<0.05) and 78 (p<0.004) in seronegatives and week 4 (p<0.0005) in seropositive subject. There was a significant difference in the percentage of Perforin CD8+ T-cells at weeks 42 (p<0.04), 52 (p<0.02) and 78 (p<0.006).

Mentions: At baseline, the majority of Ad5-specific CD8+ T-cells produced predominantly the effector functions MIP1α and perforin in both seropositive and seronegative subjects (Fig. 4A). The percentage of Ad5-specific CD8+ T-cells producing TNFα was significantly higher in baseline seropositive subjects prior to vaccination (p<0.001) but there were no differences between the groups for IL-2, IFN-γ, MIP1α or perforin.


Vaccination with Ad5 vectors expands Ad5-specific CD8 T cells without altering memory phenotype or functionality.

Hutnick NA, Carnathan DG, Dubey SA, Cox KS, Kierstead L, Makadonas G, Ratcliffe SJ, Lasaro MO, Robertson MN, Casimiro DR, Ertl HC, Betts MR - PLoS ONE (2010)

Ad specific T-cell functionality following vaccination.Ten seronegative (Ad5 nAb titer ≤18, five weeks 0–4, five weeks 0–78, grey circles) and five seropositive subjects (Ad5 nAb titer >18, white circles) received Merck Ad5 gag/pol/nef as described in Methods. Black lines recommend the mean. Grey asterics represent a significant increase from baseline in seronegative subjects and black asterics represent a significant difference from baseline in seropositive subjects. Black bars represent a significant difference between the serogroups at that time point A) Percent of the total Ad-specific response producing each cytokine. The total Ad-specific CD8+ response was computed by summing cells making at least IL-2, IFN-γ, MIP1α, or TNFα as measured by flow cytometry. The percentage of the total Ad-specific response was then computed for each cytokine. The percentage of the total response consisting of TNFα was significantly higher in seropositive subjects (p<0.0005) at baseline. B) Percentage of IL-2+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seronegative subjects at week 42 (p<0.01) and seropositive subjects at week 4(p = 0.015). C) Percentgae of TNFα+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seropositive subjects at week 52 (p<0.01) and 78 (p<0.02) and serpositive subjects at week 4 (p<0.01). There was a significant difference at baseline in the percentage of TNFα+ CD8+ T-cells between serogroups (p<0.05). D) Percentage of IFN-γ+ Ad-specific CD8+ T-cells. The percentage of IFN-γ+ CD8+T-cells was significantly increased above baseline in seronegative subjects at weeks 42 (p<0.004), 52 (p<0.003), and 78 (0.009) and in seropositives at weeks (4 (p<0.0001), 8 (p<0.04), 30 (p<0.01) and 42 (p<0.04). There was a significant difference in the percentage of CD8+IFN-γ+ CD8+ T-cells at week 78 between the serogroups (p<0.05). E) Percentage of MIP1α+ Ad-specific CD8+ T-cells. Seronegative subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells above baseline at weeks 26 (p<0.03), 42 (p<0.001), 52 (p<0.05) and 78 (p<0.004). Seropositive subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells at week 4 (p<0.0005) compared with baseline There was a significant difference in the percentage of MIP1α+CD8+ T-cells between the serogroups at weeks 30 (p<0.04), 42 (p<0.012), 52 (p<0.005), and 78 (p<0.001). F) Percentage of perforin+ Ad-specific CD8+ T-cells. The percentage of perforin+CD8+ T-cells was significantly increased above baseline at weeks 26 (p<0.02), 42 (p<0.001) 52 (p<0.05) and 78 (p<0.004) in seronegatives and week 4 (p<0.0005) in seropositive subject. There was a significant difference in the percentage of Perforin CD8+ T-cells at weeks 42 (p<0.04), 52 (p<0.02) and 78 (p<0.006).
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pone-0014385-g004: Ad specific T-cell functionality following vaccination.Ten seronegative (Ad5 nAb titer ≤18, five weeks 0–4, five weeks 0–78, grey circles) and five seropositive subjects (Ad5 nAb titer >18, white circles) received Merck Ad5 gag/pol/nef as described in Methods. Black lines recommend the mean. Grey asterics represent a significant increase from baseline in seronegative subjects and black asterics represent a significant difference from baseline in seropositive subjects. Black bars represent a significant difference between the serogroups at that time point A) Percent of the total Ad-specific response producing each cytokine. The total Ad-specific CD8+ response was computed by summing cells making at least IL-2, IFN-γ, MIP1α, or TNFα as measured by flow cytometry. The percentage of the total Ad-specific response was then computed for each cytokine. The percentage of the total response consisting of TNFα was significantly higher in seropositive subjects (p<0.0005) at baseline. B) Percentage of IL-2+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seronegative subjects at week 42 (p<0.01) and seropositive subjects at week 4(p = 0.015). C) Percentgae of TNFα+ Ad-specific CD8+ T-cells. There was a significant increase above baseline in seropositive subjects at week 52 (p<0.01) and 78 (p<0.02) and serpositive subjects at week 4 (p<0.01). There was a significant difference at baseline in the percentage of TNFα+ CD8+ T-cells between serogroups (p<0.05). D) Percentage of IFN-γ+ Ad-specific CD8+ T-cells. The percentage of IFN-γ+ CD8+T-cells was significantly increased above baseline in seronegative subjects at weeks 42 (p<0.004), 52 (p<0.003), and 78 (0.009) and in seropositives at weeks (4 (p<0.0001), 8 (p<0.04), 30 (p<0.01) and 42 (p<0.04). There was a significant difference in the percentage of CD8+IFN-γ+ CD8+ T-cells at week 78 between the serogroups (p<0.05). E) Percentage of MIP1α+ Ad-specific CD8+ T-cells. Seronegative subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells above baseline at weeks 26 (p<0.03), 42 (p<0.001), 52 (p<0.05) and 78 (p<0.004). Seropositive subjects had a significantly increased percentage of MIP1α+ CD8+ T-cells at week 4 (p<0.0005) compared with baseline There was a significant difference in the percentage of MIP1α+CD8+ T-cells between the serogroups at weeks 30 (p<0.04), 42 (p<0.012), 52 (p<0.005), and 78 (p<0.001). F) Percentage of perforin+ Ad-specific CD8+ T-cells. The percentage of perforin+CD8+ T-cells was significantly increased above baseline at weeks 26 (p<0.02), 42 (p<0.001) 52 (p<0.05) and 78 (p<0.004) in seronegatives and week 4 (p<0.0005) in seropositive subject. There was a significant difference in the percentage of Perforin CD8+ T-cells at weeks 42 (p<0.04), 52 (p<0.02) and 78 (p<0.006).
Mentions: At baseline, the majority of Ad5-specific CD8+ T-cells produced predominantly the effector functions MIP1α and perforin in both seropositive and seronegative subjects (Fig. 4A). The percentage of Ad5-specific CD8+ T-cells producing TNFα was significantly higher in baseline seropositive subjects prior to vaccination (p<0.001) but there were no differences between the groups for IL-2, IFN-γ, MIP1α or perforin.

Bottom Line: Adenoviral (Ad) vaccine vectors represent both a vehicle to present a novel antigen to the immune system as well as restimulation of immune responses against the Ad vector itself.Peripheral blood Ad5-specific CD8(+) T-cell numbers expanded after Ad5-HIV vaccination in all subjects, but differential expansion kinetics were noted in some baseline Ad5-neutralizing antibody (Ad5 nAb) seronegative subjects compared to baseline Ad5 nAb seropositive subjects.These data indicate that repeat Ad5-vector administration in humans expands Ad5-specific CD8(+) T-cells without overtly affecting their functional capacity or phenotypic properties.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Center for AIDS Research, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America.

ABSTRACT

Background: Adenoviral (Ad) vaccine vectors represent both a vehicle to present a novel antigen to the immune system as well as restimulation of immune responses against the Ad vector itself. To what degree Ad-specific CD8(+) T cells are restimulated by Ad vector vaccination is unclear, although such knowledge would be important as vector-specific CD8(+) T cell expansion could potentially further limit Ad vaccine efficacy beyond Ad-specific neutralizing antibody alone.

Methodology/principal findings: Here we addressed this issue by measuring human Adenovirus serotype 5 (Ad5)-specific CD8(+) T cells in recipients of the Merck Ad5 HIV-1 vaccine vector before, during, and after vaccination by multicolor flow cytometry. Ad5-specific CD8(+) T-cells were detectable in 95% of subjects prior to vaccination, and displayed primarily an effector-type functional profile and phenotype. Peripheral blood Ad5-specific CD8(+) T-cell numbers expanded after Ad5-HIV vaccination in all subjects, but differential expansion kinetics were noted in some baseline Ad5-neutralizing antibody (Ad5 nAb) seronegative subjects compared to baseline Ad5 nAb seropositive subjects. However, in neither group did vaccination alter polyfunctionality, mucosal targeting marker expression, or memory phenotype of Ad5-specific CD8(+) T-cells.

Conclusions: These data indicate that repeat Ad5-vector administration in humans expands Ad5-specific CD8(+) T-cells without overtly affecting their functional capacity or phenotypic properties. This is a secondary analysis of samples collected during the 016 trial. Results of the Merck 016 trial safety and immunogenicity have been previously published in the journal of clinical infectious diseases [1].

Trial registration: ClinicalTrials.gov NCT00849680[http://www.clinicaltrials.gov/show/NCT00849680].

Show MeSH
Related in: MedlinePlus