The fusion αDEC-NS1 and αDCIR2-NS1 mAbs bind to the DC DEC205 and DCIR2 receptors, respectively.

<p>(A) A density of 10⁵ CHO cells expressing either DEC205 or DCIR2 receptors were incubated on ice with 10, 1 or 0.1 µg/mL of αDEC-NS1 or αDCIR2-NS1 fusion antibodies. Binding was detected on 30.000 cells using an anti-mouse IgG1 antibody. (B) A density of 5×10⁶ splenocytes were incubated on ice with the same concentrations of the fusion antibodies described in (A). Gates on CD11c⁺CD8α⁺ (DEC205 rich population) or CD11c⁺CD8α⁻ (DCIR2 rich population) are shown in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002330#pntd.0002330.s001" target="_blank">figure S1</a>. Binding was detected on 1×10⁶ cells using an anti-mouse IgG1 antibody. One experiment representative of three is shown.</p

CD4⁺ and CD8⁺ T cells play important roles on the protective responses induced in mice immunized with the αDEC-NS1 mAb.

<p>Mice were immunized as described in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002330#pntd-0002330-g003" target="_blank">figure 3</a>. One day before the challenge, mice were intravenously injected with 100 µg of αCD4 or αCD8 purified antibodies to deplete the respective T cell populations. Survival (A) and morbidity (B) were monitored daily for 21 days after the challenge. n = 8 mice/group. * Refers to p<0.05 when compared to the αDEC-NS1+αCD4 mAb, adjuvant and saline groups.</p

Induction of anti-NS1 antibody responses in mice immunized with the fusion αDEC-NS1 and αDCIR2-NS1 mAbs.

<p>(A) Immunization protocol describing the order of immunizations, bleedings and challenge. Mice were immunized i.p. with 2 doses of 5 µg of each fusion antibody in the presence of 50 µg of poly (I:C) or 10 µg of recombinant NS1 co-administered with 50 µg of poly (I:C). (B) Anti-NS1 antibody titers 4 days before (white symbols) or 14 days after (black symbols) the administration of the second dose. (C) Anti-NS1 antibody titers obtained 14 days after the second dose were detected using intact (white symbols) or boiled (black symbols) NS1 protein. Symbols represent individual mice and horizontal lines represent the mean value for each group. Fusion antibodies used for immunizations are depicted below. Data were analyzed by a one-way ANOVA followed by the post-test HSD Tukey. P-value indicators * and *** refer to p<0.05 and p<0.01, respectively, while ns = not significant.</p