Article Figures & Data
Figures
- FIG. 1.
Frequencies of Gag-specific IFN-γ-secreting cells from monkeys immunized with Ad5-gag. These values are expressed as the number of SFC per 106 PBMCs. Naïve monkeys (adenovirus type 5 neutralization (neut) titers < 10) were immunized with various doses of Ad5-gag at weeks 0 and 24. Monkeys that were pretreated with one or three 1010 viral particle (vp) doses of non-Gag-encoding adenovirus type 5 virus (producing neutralization titers of ≈30 to 90 and ≈270 to 810, respectively, at the time of the first Ad5-gag injection) were immunized with 1011 viral particles of Ad5-gag at weeks 0 and 24. PBMCs collected at weeks 0, 4, 24, and 28 were incubated in the absence (mock) or presence of the HIV-1 Gag peptide pool, and the mock-corrected response levels are shown here for each monkey. Mock-treated background responses averaged 3 SFC per 106 PBMCs (standard deviation, ≈5 SFC per 106 PBMCs).
- FIG. 2.
CTL responses in monkeys immunized with Ad5-gag. Peptide-pulsed autologous BLCLs were used as target cells at the indicated effector-to-target cell (E:T) ratios, treated with PBMCs (collected at week 8) from Ad5-gag vaccinees restimulated for 2 weeks with gag-expressing vaccinia virus. The percentages of lysed target cells are shown for unpulsed cells (open squares) and cells pulsed with the HIV-1 Gag peptide pool (solid diamonds).
- FIG. 3.
Time course of induction of Gag-specific T-cell responses from PBMCs of representative monkeys that received 1011 viral particles and 109 viral particles of Ad5-gag at week 0 and week 24 (A). Mock responses were subtracted to give the reported levels shown here. Kinetics of induction of p24-specific antibodies as a function of the Ad5-gag dose and adenovirus type 5 preexposure (B). The geometric mean titers (GMT) are shown with standard errors for each cohort of either three or six monkeys. Each curve is defined by the Ad5-gag dose (viral particles, vp) and adenovirus type 5 neutralization titers measured at the start of the immunization. Titers below the dection limit of 10 mMU/ml are scored at 5 mMU/ml (line).
- FIG. 4.
Intracellular IFN-γ staining of PBMCs from Ad5-gag vaccinees following overnight incubation in medium alone (mock) or in medium plus the Gag peptide pool (Gag 20-aa). CD3+ lymphocytes are shown and were characterized for CD8+ staining and IFN-γ production. Numbers reflect the percentage of CD3+ lymphocytes that were CD8+ IFN-γ+ (upper right quadrant) or CD4+ IFN-γ+ (lower right quadrant).
- FIG. 5.
Frequencies of Gag-specific IFN-γ-secreting cells from monkeys immunized with Ad5-gag and MVA-gag. Priming immunizations were administered at weeks 0 and 4, followed by a booster shot at week 24 with the same virus. Shown are the mock-corrected levels against the Gag peptide pool prior to the first dose (pre), 4 weeks after the second dose (post prime), time of the boost (preboost), and 4 weeks after the viral boost (postboost) for each animal.
- FIG. 6.
Frequencies of Gag-specific IFN-γ-secreting cells from PBMCs of macaques immunized following various DNA prime-adenovirus type 5 boost regimens. (A) Macaques were given multiple doses of 5 mg of V1Jns-gag DNA intramuscularly in phosphate-buffered saline, with aluminum phosphate (alum) or with CRL1005 (7.5 mg) at weeks 0, 4, and 8. Shown are levels of antigen-specific T cells (mock corrected) at different time points during and after the priming immunizations. (B) Macaques were treated intramuscularly with three doses (weeks 0, 4, and 8) of 5 mg of V1Jns-gag with and without adjuvants (CRL1005 at 7.5 or 22.5 mg) and boosted with an intramuscular dose of Ad5-gag (107 viral particles or 1011 viral particles) at weeks 24 to 26. Shown are the mock-corrected levels prior to the first dose (pre), 4 weeks after the third dose (post prime), time of the boost (preboost), and 4 weeks after the viral boost (postboost) for each animal.
- FIG. 7.
Intracellular IFN-γ staining of PBMCs from a representative cohort of macaques immunized with three doses of V1jns-gag formulated with CRL1005-based adjuvant (weeks 0, 4, and 8) followed by 107 viral particles of Ad5-gag boost (week 24). Samples were collected at week 10 (postprime) and week 28 (postboost). Shown are CD3+ lymphocytes following incubation with the HIV-1 Gag peptide pool that were characterized for CD8+ staining and IFN-γ production. For all samples, intracellular IFN-γ production in the absence of the peptide pool was minimal (data not shown). Numbers reflect the percentage of CD3+ lymphocytes that were CD8+ IFN-γ+ (upper right quadrant) or CD4+ IFN-γ+ (lower right quadrant) after subtraction of the mock levels.
- FIG. 8.
Percentages of Gag-specific T cells that are CD3+ CD8+ in rhesus macaques immunized with various regimens. The adenovirus type 5 prime-adenovirus type 5 boost (Ad5-Ad5) cohort consisted of animals given two priming doses of the vector (109 or 1011 viral particles) followed by a booster shot of the same dose. The DNA-CRL1005 prime-adenovirus type 5 boost cohort comprised animals given three priming immunizations with 5 mg of V1Jns-gag formulated with CRL1005-based adjuvant, followed by 107 viral particles of Ad5-gag. The percentage values are reported for this cohort of 11 animals before (DNA-CRL1005) and after the boost (DNA-CRL1005-Ad5). The dark diamonds and bars are the cohort arithmetic means and standard errors of the mean, respectively. These data points include those for the animals in Fig. 4 and 7.
