Figures & data
Relative binding antibody (bAb) levels against whole ZIKV and DENV-2 were examined using ELISA. The kinetics of binding IgM in serum that was collected longitudinally from (a) DENV-2 only (ZIKV naïve) infected animals (n = 4), (b) ZIKV-infected animals prior to DENV-2 infection (n = 5). The kinetics of binding IgG in serum that was collected longitudinally from (c) DENV-2 only (ZIKV naïve) infected animals (n = 4), (d) ZIKV-infected animals prior to DENV-2 infection (n = 5). Statistical significance was determined using multiple unpaired t-tests and corrected for multiple comparisons using the Holm-Sidak method. A p < 0.05 was considered significant and * indicates significant difference between groups
![Fig. 1 Kinetics of cross-reactive antibody responses.Relative binding antibody (bAb) levels against whole ZIKV and DENV-2 were examined using ELISA. The kinetics of binding IgM in serum that was collected longitudinally from (a) DENV-2 only (ZIKV naïve) infected animals (n = 4), (b) ZIKV-infected animals prior to DENV-2 infection (n = 5). The kinetics of binding IgG in serum that was collected longitudinally from (c) DENV-2 only (ZIKV naïve) infected animals (n = 4), (d) ZIKV-infected animals prior to DENV-2 infection (n = 5). Statistical significance was determined using multiple unpaired t-tests and corrected for multiple comparisons using the Holm-Sidak method. A p < 0.05 was considered significant and * indicates significant difference between groups](/cms/asset/2f5e7359-bf69-439f-8026-9cd4ae103322/temi_a_12040039_f0001_ob.png)
Percentage neutralization of (a) ZIKV and (b) DENV-2 by serum that was collected longitudinally from DENV-2 only (ZIKV naïve) (n = 4) and ZIKV-infected animals prior to DENV-2 infection (n = 5). Line represents PRNT50 titres
![Fig. 2 ZIKV-infected animals display little or no cross-neutralizing antibody responses against DENV-2.Percentage neutralization of (a) ZIKV and (b) DENV-2 by serum that was collected longitudinally from DENV-2 only (ZIKV naïve) (n = 4) and ZIKV-infected animals prior to DENV-2 infection (n = 5). Line represents PRNT50 titres](/cms/asset/83bc7a93-85c9-42ab-a5c5-9f86104ff25b/temi_a_12040039_f0002_ob.png)
Kinetics of neutralizing antibody responses against DENV-2 using serum that was collected longitudinally from DENV-2 only (ZIKV naïve) and ZIKV-infected animals at (a) day 0, (b) day 1, (c) day 4, (d) day 7, (e) day 14, and (f) day 56 after DENV-2 infection (n = 5). Line represents 50% neutralization
![Fig. 3 ZIKV immune animals display delayed anamnestic cross-neutralizing antibody responses after DENV-2 infection.Kinetics of neutralizing antibody responses against DENV-2 using serum that was collected longitudinally from DENV-2 only (ZIKV naïve) and ZIKV-infected animals at (a) day 0, (b) day 1, (c) day 4, (d) day 7, (e) day 14, and (f) day 56 after DENV-2 infection (n = 5). Line represents 50% neutralization](/cms/asset/3b4dc73b-9fc4-4b42-a93d-1ade65f2f54f/temi_a_12040039_f0003_ob.png)
Kinetics of DENV-2 viremia
Kinetics of binding antibody (bAb) responses against DENV-2 in serum showing relative levels of (a) IgM and (b) IgG against DENV-2 using serum that was collected longitudinally from ZIKV-infected animals 1, 4, and 7 days after DENV-2 infection (n = 5). Percentage OD450 values relative to each animals day 56 post-ZIKV infection (day 0 DENV-2) values are shown. Line represents day 0 vales. Statistical significance was determined using multiple unpaired t-tests and corrected for multiple comparisons using the Holm-Sidak method. A p < 0.05 was considered significant and * indicates significant difference between groups
![Fig. 4 ZIKV infected animals display delayed binding antibody responses after DENV-2 infection.Kinetics of binding antibody (bAb) responses against DENV-2 in serum showing relative levels of (a) IgM and (b) IgG against DENV-2 using serum that was collected longitudinally from ZIKV-infected animals 1, 4, and 7 days after DENV-2 infection (n = 5). Percentage OD450 values relative to each animals day 56 post-ZIKV infection (day 0 DENV-2) values are shown. Line represents day 0 vales. Statistical significance was determined using multiple unpaired t-tests and corrected for multiple comparisons using the Holm-Sidak method. A p < 0.05 was considered significant and * indicates significant difference between groups](/cms/asset/089ddf76-6a8b-4488-8dcd-2f38197d475a/temi_a_12040039_f0004_ob.png)
Relative levels of (a) DENV-2 and (b) ZIKV binding IgG1, IgG2, and IgG3 in serum that was collected at day 14 and day 56 post-infection from DENV-2 only (n = 4) infected animals. Relative levels of (c) DENV-2 and (d) ZIKV binding IgG1, IgG2, and IgG3 in serum that was collected at day 14 and day 56 post-infection from ZIKV-infected animals prior to DENV-2 infection (n = 5). Relative levels of (e) DENV-2 and (f) ZIKV binding IgG1, IgG2, and IgG3 in serum that was collected at day 1, 4, 7, and 56 post-infection from ZIKV-infected animals after DENV-2 infection (n = 5). Statistical significance was determined using one-way ANOVA and differences between time-points were determined by post-hoc analysis using Tukey’s multiple comparisons test. A p < 0.05 was considered significant. Error bars represent standard error
![Fig. 5 IgG1 is the primary subclass of IgG in serum from ZIKV naïve and infected animals prior to and after infection with DENV-2.Relative levels of (a) DENV-2 and (b) ZIKV binding IgG1, IgG2, and IgG3 in serum that was collected at day 14 and day 56 post-infection from DENV-2 only (n = 4) infected animals. Relative levels of (c) DENV-2 and (d) ZIKV binding IgG1, IgG2, and IgG3 in serum that was collected at day 14 and day 56 post-infection from ZIKV-infected animals prior to DENV-2 infection (n = 5). Relative levels of (e) DENV-2 and (f) ZIKV binding IgG1, IgG2, and IgG3 in serum that was collected at day 1, 4, 7, and 56 post-infection from ZIKV-infected animals after DENV-2 infection (n = 5). Statistical significance was determined using one-way ANOVA and differences between time-points were determined by post-hoc analysis using Tukey’s multiple comparisons test. A p < 0.05 was considered significant. Error bars represent standard error](/cms/asset/34bae482-332c-4cd7-afec-7a10643cdaa1/temi_a_12040039_f0005_ob.png)
Fold enhancement of DENV-2 infection in K562 cells using serum that was collected (a) from ZIKV-infected animals (n = 5) prior to DENV-2 infection and (b) at day 56 after DENV-2 infection. Statistical significance was determined using One-way ANOVA and differences between time-points were determined by post-hoc analysis using Tukey’s multiple comparisons test. A p < 0.05 was considered significant. (c) Percentage neutralization of DENV-2 using serum that was collected at day 56 post-infection from animals that was simultaneously infected with ZIKV and DENV-2 (n = 5). (d) Fold enhancement of DENV-2 infection in K562 cells using serum that was collected at 56 days from rhesus macaques that were simultaneously infected with ZIKV and DENV-2 (n = 5). Statistical significance was determined using One-way ANOVA and differences between time-points were determined by post-hoc analysis using Tukey's multiple comparisons test. A p < 0.05 was considered significant. Error bars represent standard error
![Fig. 6 Serum from ZIKV infected animals collected after infection with DENV-2 demonstrate antibody dependent enhancement at a dilution of 1:100.Fold enhancement of DENV-2 infection in K562 cells using serum that was collected (a) from ZIKV-infected animals (n = 5) prior to DENV-2 infection and (b) at day 56 after DENV-2 infection. Statistical significance was determined using One-way ANOVA and differences between time-points were determined by post-hoc analysis using Tukey’s multiple comparisons test. A p < 0.05 was considered significant. (c) Percentage neutralization of DENV-2 using serum that was collected at day 56 post-infection from animals that was simultaneously infected with ZIKV and DENV-2 (n = 5). (d) Fold enhancement of DENV-2 infection in K562 cells using serum that was collected at 56 days from rhesus macaques that were simultaneously infected with ZIKV and DENV-2 (n = 5). Statistical significance was determined using One-way ANOVA and differences between time-points were determined by post-hoc analysis using Tukey's multiple comparisons test. A p < 0.05 was considered significant. Error bars represent standard error](/cms/asset/a9315cd4-0522-4fc2-acd3-7e31eb878573/temi_a_12040039_f0006_ob.png)