A Phase I Double Blind, Placebo-Controlled, Randomized Study of a Multigenic HIV-1 Adenovirus Subtype 35 Vector Vaccine in Healthy Uninfected Adults

<div><h3>Background</h3><p>We conducted a phase I, randomized, double-blind, placebo-controlled trial to assess the safety and immunogenicity of escalating doses of two recombinant replication defective adenovirus serotype 35 (Ad35) vectors containing gag, reverse transcriptase, integrase and nef (Ad35-GRIN) and env (Ad35-ENV), both derived from HIV-1 subtype A isolates. The trial enrolled 56 healthy HIV-uninfected adults.</p> <h3>Methods</h3><p>Ad35-GRIN/ENV (Ad35-GRIN and Ad35-ENV mixed in the same vial in equal proportions) or Ad35-GRIN was administered intramuscularly at 0 and 6 months. Participants were randomized to receive either vaccine or placebo (10/4 per group, respectively) within one of four dosage groups: Ad35-GRIN/ENV 2×10⁹ (A), 2×10¹⁰ (B), 2×10¹¹ (C), or Ad35-GRIN 1×10¹⁰ (D) viral particles.</p> <h3>Results</h3><p>No vaccine-related serious adverse event was reported. Reactogenicity events reported were dose-dependent, mostly mild or moderate, some severe in Group C volunteers, all transient and resolving spontaneously. IFN-γ ELISPOT responses to any vaccine antigen were detected in 50, 56, 70 and 90% after the first vaccination, and in 75, 100, 88 and 86% of Groups A–D vaccine recipients after the second vaccination, respectively. The median spot forming cells (SFC) per 10⁶ PBMC to any antigen was 78–139 across Groups A–C and 158–174 in Group D, after each of the vaccinations with a maximum of 2991 SFC. Four to five HIV proteins were commonly recognized across all the groups and over multiple timepoints. CD4+ and CD8+ T-cell responses were polyfunctional. Env antibodies were detected in all Group A–C vaccinees and Gag antibodies in most vaccinees after the second immunization. Ad35 neutralizing titers remained low after the second vaccination.</p> <h3>Conclusion/Significance</h3><p>Ad35-GRIN/ENV reactogenicity was dose-related. HIV-specific cellular and humoral responses were seen in the majority of volunteers immunized with Ad35-GRIN/ENV or Ad35-GRIN and increased after the second vaccination. T-cell responses were broad and polyfunctional.</p> <h3>Trial Registration</h3><p>ClinicalTrials.gov <a href="http://clinicaltrials.gov/ct2/results?term=NCT00851383">NCT00851383</a></p> </div

Safety and Immunogenicity Study of Multiclade HIV-1 Adenoviral Vector Vaccine Alone or as Boost following a Multiclade HIV-1 DNA Vaccine in Africa

We conducted a double-blind, randomized, placebo-controlled Phase I study of a recombinant replication-defective adenovirus type 5 (rAd5) vector expressing HIV-1 Gag and Pol from subtype B and Env from subtypes A, B and C, given alone or as boost following a DNA plasmid vaccine expressing the same HIV-1 proteins plus Nef, in 114 healthy HIV-uninfected African adults.Volunteers were randomized to 4 groups receiving the rAd5 vaccine intramuscularly at dosage levels of 1×10(10) or 1×10(11) particle units (PU) either alone or as boost following 3 injections of the DNA vaccine given at 4 mg/dose intramuscularly by needle-free injection using Biojector® 2000. Safety and immunogenicity were evaluated for 12 months. Both vaccines were well-tolerated. Overall, 62% and 86% of vaccine recipients in the rAd5 alone and DNA prime - rAd5 boost groups, respectively, responded to the HIV-1 proteins by an interferon-gamma (IFN-γ) ELISPOT. The frequency of immune responses was independent of rAd5 dosage levels. The highest frequency of responses after rAd5 alone was detected at 6 weeks; after DNA prime - rAd5 boost, at 6 months (end of study). At baseline, neutralizing antibodies against Ad5 were present in 81% of volunteers; the distribution was similar across the 4 groups. Pre-existing immunity to Ad5 did not appear to have a significant impact on reactogenicity or immune response rates to HIV antigens by IFN-γ ELISPOT. Binding antibodies against Env were detected in up to 100% recipients of DNA prime - rAd5 boost. One volunteer acquired HIV infection after the study ended, two years after receipt of rAd5 alone.The HIV-1 rAd5 vaccine, either alone or as a boost following HIV-1 DNA vaccine, was well-tolerated and immunogenic in African adults. DNA priming increased the frequency and magnitude of cellular and humoral immune responses, but there was no effect of rAd5 dosage on immunogenicity endpoints.ClinicalTrials.gov NCT00124007

Demographic Characteristics of the Study Population.

<p>Demographic Characteristics of the Study Population.</p

Magnitude of HIV-specific antibodies.

<p>The geometric mean of a) anti-ENV Subtype A (UG037)- and b) anti-p24 (IIIB)-specific antibody titers is shown at baseline, at 4 and 24 weeks post-first vaccination, and at 2, 14, 28, 40 and 48 weeks post-second vaccination.</p

CONSORT Flow Diagram.

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

Polychromatic flow cytometry of Env-specific T cells.

a<p>All samples at baseline,</p>b<p>Samples at 2 weeks post second vaccination,</p>c<p>Frequency of positive cells.</p

Polychromatic flow cytometry of GRIN-specific T cells.

a<p>All samples at baseline,</p>b<p>Samples at 2 weeks post second vaccination,</p>c<p>Frequency of positive cells.</p

Time Course of Local and Systemic Reactions Post First (Vac1) and Post Second (Vac2) Vaccination per Maximum Severity Assessment for Placebo, Group A (2×10⁹ vp), Group B (2×10¹⁰ vp), Group C (2×10¹¹ vp) and Group D (1×10¹⁰ vp).

<p>The Y-axis of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041936#pone-0041936-g002" target="_blank">Figure 2</a> represents the number of volunteers experiencing reactogenicity events (panel A for local reactions and panel B for systemic reactions post first and second vaccinations, upper and lower rows respectively) for each group, while the X-axis represents the days of occurrence of the events, Day 0 being the day of vaccination. Volunteers did a self-assessment of reactogenicity with a memory card on Day 0 (evening of vaccination) and daily through Day 13, reviewed by the investigator at Days 3, 7 and 14. The figure shows the maximum severity assessment grade recorded as per the volunteer’s and clinic’s assessments combined. The severity grade of the reactogenicity events is indicated by color codes (mild: yellow; moderate: orange; severe: red).</p

Number of Volunteers Enrolled, Vaccination Schedule, Sample Collection and Assessment time points.

<p>W  =  week, W0 and W24 are vaccination visits.</p>a<p>(V/P) Number of Vaccine recipients/number of Placebo recipients per group.</p>b<p>Screen window up to 65 days prior to enrollment for anti-Ad35 antibodies.</p>c<p>Serum neutralizing antibodies against Ad35.</p>d<p>Vaccine-induced HIV-1 specific IFN-γ ELISPOT responses.</p>e<p>Vaccine-induced HIV-1 specific humoral immune responses (Env and p24 Gag ELISA).</p>f<p>Polychromatic Flow Cytometry.</p