Fig. 5. CYV-ZIKV-mosquitoes bites protect IFNAR^−/− C57BL/6 mice from the ZIKV challenge.

a–p Mice were bitten with CYV-ZIKV infected mosquitoes (10 per mouse) three times and challenged on day 42 with 10³ FFU (>50 LD₅₀) of the ZIKV MR766 strain (f–h), 10⁵ FFU (10 LD⁵₀) of the Natal-RGN strain (j–l), or 10⁴ PFU (>5 × 10² LD₅₀) of the GZ01 strain (n-p) (n = 6). Mice bitten with naïve mosquitoes were used as controls (Con.). a Scheme of the immunization and challenge experiment. b–d Viral titers of CYV-ZIKV in the saliva of 80 mosquitoes used to immunize mice against ZIKV MR766 (b), Natal-RGN (c), or GZ01(d) were determined 12 dpi (n = 3). 1st, the first bite; 2nd, the second bite; 3rd, the third bite. Data are presented as means ± SD of triplicate measurements. Con. indicates control saliva of naïve mosquitoes. e, i, m Neutralizing antibody levels of sera on day 35 against the indicated ZIKV strain. The dashed lines indicate the 1:20 detection limit and the horizontal bars indicate the mean. e MR766, n = 5; Con., n = 4. i n = 6. m n = 6. f, j, n Survival rate of mice. g, k, o Weight change in mice. Data are presented as means ± SD. h, l, p ZIKV viremia post-challenge, measured by real-time PCR using ACTB as the reference gene. The dashed lines indicate the detection limit. Data are presented as means ± SD. Similar results were obtained in three independent experiments. Source data are provided as a Source Data file.