Article Figures & Data
Figures
- FIG. 1.
Immunogenicity of LT delivered by TCI. Following pretreatment of the skin with a nonwoven abrasive EKG prep pad to disrupt the stratum corneum, a wet patch containing 50 μg of one of three lots of LT was applied to the pretreated area on days 0 and 21. Serum samples obtained on days 0, 21, 42, 90, and 381 were analyzed to determine the presence of anti-LT IgG and IgA by ELISA. There were no significant differences between the anti-LT immune responses of the three groups (P < 0.05). The serum antibody levels for the combined treatment groups are expressed in ELISA units; the bars indicate geometric means, and the error bars indicate 95% confidence intervals. The percentages are percentages of seroconversion (a twofold increase compared with the baseline for IgG and a fourfold increase for IgA).
- FIG. 2.
LT toxin neutralization by LT immune sera. Serum samples obtained on days 0, 21, 42, 90, and 381 from subjects vaccinated on days 0 and 21 with 50 μg of LT by TCI were tested to determine their abilities to neutralize LT in a Y-1 serum neutralization assay. The serum antibody levels for the combined treatment groups are expressed as average ED50s, and the error bars indicate 95% confidence intervals.
- FIG. 3.
Comparison of neutralization of LT and CT toxins by LT immune sera. Pooled serum samples obtained on days 0 and 42 from subjects vaccinated on days 0 and 21 with 50 μg of LT by TCI were tested to determine their abilities to neutralize CT in a Y-1 serum neutralization assay. Toxin activity is indicated by the reduction in OD530, which reflects the number of neutral red-stained cells remaining after treatment. Toxin neutralization of both LT and CT by serum is shown.
- FIG. 4.
Anti-LT serum IgG induced following a live infectious challenge. Subjects received an oral infectious challenge consisting of 1 × 109 viable ETEC E2447A LT+ ST+ bacteria. Serum samples obtained on day 0, as well as on days 7 and 28 following the oral challenge, were analyzed to determine the presence of anti-LT IgG by ELISA. The serum antibody levels are expressed in ELISA units; the bars indicate geometric means, and the error bars indicate 95% confidence intervals. The percentages are percentages of seroconversion.
- FIG. 5.
Serological recognition of LT by sera from CTB-vaccinated subjects. Sera obtained from subjects 35 days after vaccination with a single dose of an oral whole-cell cholera vaccine containing 1 mg CTB and killed V. cholerae cells were analyzed to determine their recognition of LT, using an ELISA. The serum antibody levels are expressed in ELISA units; the bars indicate geometric means, and the error bars indicate 95% confidence levels. The percentage is the percentage of seroconversion.
- FIG. 6.
Correlation between net TEWL and fold increase in the IgG titer. TEWL was measured on the skin before pretreatment and after skin pretreatment but before patch application in 51 of 59 subjects at day 42. Individual net TEWL (posttreatment TEWL − baseline TEWL) at the time of vaccination is plotted against the increase in anti-LT IgG at day 42 (r = 0.59, P = 0.001).
- FIG. 7.
Biopsy of control and nonwoven abrasive pad-treated human skin. After informed consent, a shave biopsy of human skin was obtained from a control site (A) and a site that was treated with 15 strokes of a nonwoven abrasive EKG pad (B). The skin was formalin fixed and stained using hematoxylin and eosin. The three layers of the skin, the stratum corneum, epidermis, and dermis, are visible, and in the group pretreated with the nonwoven abrasive pad a modest decrease in the thickness of the stratum corneum is visible.
Tables
- TABLE 1.
LT IgG GMT for subjects immunized transcutaneously
Study day GMT (EU) Group 1 (n = 20) Group 2 (n = 20) Group 3 (n = 20) All treatments (n = 60) 0 369 656a 514 499 21 5,217 8,553 5,595 6,297 42 12,734 12,988 10,939 12,185 90 10,772 14,350 11,222 12,016 ↵ a P = 0.008 for a pairwise comparison of group 1 and group 2 using the covariate-adjusted and unadjusted generalized linear model.
- TABLE 2.
TEWL resultsa
Group n TEWL(g/m2/h) Baseline After pretreatment of skin Net n Mean (SD) Range n Mean (SD) Range n Mean (SD) Range 1 18 18 7.6 (2.2) 4.2-11.2 17 21.9 (16.1) 8.8-74.2 17 14.2 (15.6) 3.2-65.9 2 17 16 7.2 (2.1) 3.0-11.2 16 20.3 (9.4) 10.0-45.4 16 13.2 (9.2) 4.29-37.4 3 16 17 6.9 (2.1) 2.0-10.8 17 18.2 (9.3) 7.3-34.7 17 11.3 (8.7) 1.4-27.1 All 51 51 7.3 (2.1)b 2.0-11.22 50 20.1 (11.9)b 7.3-74.2 50 12.9 (11.5) 1.4-65.9 ↵ a Baseline TEWL, TEWL after pretreatment of the skin at day 42, and net TEWL were determined. The net TEWL was calculated by subtracting the baseline TEWL from the TEWL after skin pretreatment. The differences between groups for baseline TEWL, TEWL after skin pretreatment, and net TEWL were not significant (P > 0.005, as determined by a t test).
↵ b The differences between the group TEWL and the combined subject TEWL after skin pretreatment and baseline TEWL were significant (P < 0.01).
- TABLE 3.
Summary of stratum corneum cell layer measurements
Subject No. of strokes No. of stratum corneum cell layers removed (mean ± SD) % Reduction 001 None (control) 17 ± 2.0 15 12.5 ± 0.7 26 20 9.0 ± 1.5 47 002 None (control) 18.8 ± 2.2 15 14.1 ± 1.5 25 20 7.9 ± 2.1 58 003 None (control) 22.0 ± 1.8 15 14.4 ± 1.7 35 20 11.8 ± 2.4 46 Mean None (control) 19.3 15 13.7 29 20 9.6 50
