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Table 1.

Number of Volunteers Enrolled, Vaccination Schedule, Sample Collection and Assessment Time Points.

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Figure 1.

CONSORT Flow Diagram.

Number of individuals assessed for eligibility, enrolled and randomized to study vaccine(s) and respective placebo, followed-up and analyzed.

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Table 2.

Demographics, by Group Assignment.

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Figure 2.

Solicited Local and Systemic Events.

Figure 2a; Local reactogenicity and systemic signs and symptoms collected over 3 days post vaccination. Maximum severity of local reactions was significantly greater after rAd5 than after placebo (p = 0.006 and p<0.001 for rAd5 alone and rAd5 boost, respectively). For systemic signs and symptoms, the severity was significantly greater after DNA (p = 0.036) and after rAd5 boost (p = 0.028), than after the corresponding placebo. Figure 2b; Local reactogenicity and systemic events post rAd5 by dosage. Combining low dosage (LD) groups and high dosage (HD) groups, the maximum systemic reaction per volunteer post rAd5 was significantly higher (p = 0.025) in the HD group than in the LD group (41% versus 21%, respectively, were moderate or severe). Mild: open bars; Moderate: cross-hatched bars; and Severe: dark bars.

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Table 3.

Number of Unsolicited Adverse Events within 28 Days Post Any Vaccination, by Severity and Group.

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Table 4.

IFN-γ ELISPOT Response Rates at Single Post Vaccination Time Points.

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Figure 3.

Impact of DNA prime on ELISPOT responses.

Comparison of percent ELISPOT responders after rAd5 alone (Groups A and B) versus DNA prime - rAd5 boost over time (Groups C and D). Vertical lines represent 95% Confidence Intervals.

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Figure 4.

Impact of DNA prime on ELISPOT magnitude.

Values plotted are the mean background-subtracted ELISPOT counts (Spot Forming Cells, SFC) over all positive peptide responses per volunteer at that visit. Shaded bars are subjects with any positive response at 4 weeks post 3rd DNA, white boxes are those with no positive responses at this time point.

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Figure 5.

Magnitude of ELISPOT and % response rate.

The magnitude of the response was measured by SFC per million peripheral blood mononuclear cells (SFC/m PBMC). The x-axis row one shows the peptide pool, row two the % response rate and row three the vaccine group. Black dots indicate positive responses, defined by background-subtracted values greater than the cutoff, more than 3 times mean background SFC count, and a coefficient of variation of not more than 70% amongst replicate wells. The rAd5 response rates correspond to both dose groups combined. The box plots summarize positive responses only (i.e., median, 1st and 3rd quartiles, minimum/maximum). The cut-offs for EnvA, EnvB, Gag, Nef, PolB1 and PolB2, as determined by the level of non-specific responses from at least 180 samples from unvaccinated individuals, are 40, 51, 54, 68, 51 and 38 respectively. The Pol response is the maximum of PolB1 and PolB2 and positive if either one is positive.

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Table 5.

Percent of Volunteers with Positive IFN-γ ELISPOT Response to Vaccine Antigens.

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Table 6.

ICS Responses Post DNA Prime - rAd5 Boost.

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Figure 6.

Induction of CD8 T cell-mediated inhibition of HIV-1-IIIB replication following vaccination with a DNA prime - rAd5 boost or rAd5 alone regimen.

Viral inhibition was assessed in placebo recipients and prior to any vaccination (•); following DNA alone (▪); DNA prime - rAd5 boost at 6 and 12 weeks (▴and ▵respectively) and after rAd5 alone at 6 and between 36 to 48 weeks (♦ and ⋄ respectively). The 1.13 log10 inhibition value above which inhibition is considered HIV-1 specific is indicated by the hashed line. Medians for all groups are indicated (—).

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Figure 7.

HIV antibody titers.

Distribution of HIV-specific antibody titers at 1 month post rAd5 alone (Groups A and B) and rAd5 boost (Groups C and D) in vaccine recipients, by protein and treatment group. The Y-axis shows the antibody titer on a log scale with box plots showing the median, 1st and 3rd quartiles.

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