Figure 3.

Post-TAK-003 vaccination sera protect against dengue virus (DENV)-2 NS1-induced hyperpermeability, and protection correlates with IgG concentration. The effect of pre- or postvaccination sera on DENV-2 NS1-induced endothelial hyperpermeability was evaluated by TEER. Representative graphs are shown for (A) DENV-naive (subject ID: 1023014) and (B) DENV-preimmune (subject ID: 1052010) samples. HPMEC were grown on Transwell semipermeable membranes (0.4 µm pore size), and serum samples (30 µL) were added to the apical chamber in the presence or absence of 5 µg/mL DENV-2 NS1 (DENV-2 NS1, blue squares; day 0 serum alone, green diamonds; day 120 serum alone, red triangles; day 0 serum + DENV-2 NS1, inverted pink triangles; day 120 serum + DENV-2 NS1, light blue ×). Endothelial permeability was measured at indicated time points over 48 hours. Dotted line indicates change of medium. Relative TEER values from 1 independent experiment performed in duplicate are plotted. Error bars indicate SEM. C, Correlation analysis of 6 DENV-naive and 6 preimmune samples from both day 0 and day 120 (24 total samples). Area under the curve (AUC) values were calculated for each curve in each TEER experiment. The absolute reduction in relative TEER for each serum sample (A, B, and Supplementary Figures 3 and 4) was calculated by subtracting the AUC of NS1 + serum from the AUC of NS1 alone. Reduction was plotted against the log₁₀ of IgG concentrations for each sample, and a correlation analysis was performed. The dotted line represents the reduction of TEER by DENV-positive (+) control serum. Naive subjects, day 0 (filled) and day 120 (open) diamonds; preimmune subjects, day 0 (filled) and day 120 (open) squares; positive control, open black circle. Abbreviations: Ab, antibody; DENV-nv-TAK-003, DENV-naive TAK-003 recipients; DENV-pre-TAK-003, DENV-preimmune TAK-003 recipients; HPMEC, human pulmonary microvascular endothelial cells; TEER, transendothelial electrical resistance.