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. 2017 Nov 27;13(11):e1006743. doi: 10.1371/journal.ppat.1006743

Fig 6. Vaccination with a WNV E-αCD180 conjugate protects BAFFR-/- mice from WNV infection and enhances WNV E-specific Abs responses.

Fig 6

A-D, WT and BAFFR-/- mice were inoculated i.v. with PBS, 20 μg/mouse of WNV E- anti-CD180 protein conjugate (WNVE-αCD180) or WNV E-ratIgG2a (WNVE-iso) 30 days prior to s.c. infection with WNV (102 PFU). Mice were monitored daily for survival (A) and clinical signs of disease (B). A, statistics were performed using a log-rank test for significance. B, the multiple t test Holm-Sidak method was used to determine significance. A and B, graphs summarize data from >4 independent experiments, N = 14 for WT mice given PBS, N = 6 for WT mice immunized with WNV E-αCD180, N = 17 for BAFFR-/- mice given PBS, N = 23 for BAFFR-/- mice immunized with WNV E-iso, N = 37 for BAFFR-/- mice immunized with WNV E-αCD180. Anti-WNV E IgM and IgG measured by ELISA (C) and anti-WNV nAbs measured by PRNT (D) in sera harvested 8 days after WNV infection. C and D, graphs summarize data from three independent experiments, N = 9 for WT mice given PBS, N = 6 for WT mice immunized with WNV E-αCD180, N = 9 for BAFFR-/- mice given PBS, N = 12 for BAFFR-/- mice immunized with WNV E-iso, N = 12 for BAFFR-/- mice immunized with WNV E-αCD180. In C and D statistics were determined by Kruskal-Wallis corrected with Dunn’s multiple comparisons post-test. E, WT, BAFFR-/- and μMT mice were inoculated i.v. with 20 μg/mouse WNV E-αCD180 30 days prior s.c. infection with WNV. Survival data are from two independent experiments (N = 10). Statistics were determined using the log-rank test for significance. F, Heat-inactivated sera obtained from naïve BAFFR-/- mice (open circles), WT mice vaccinated with WNVE-αCD180 (triangles) or from BAFFR-/- mice vaccinated with WNVE-αCD180 (closed circles) and challenged with WNV (sera obtained between day5 and day 28 post-infection), were inoculated into μMT mice 1 day prior and 1 day after WNV infection (See Methods) and mice were monitored for survival. Survival data are from one experiment (mice receiving either naïve BAFFR-/- sera or immune WT sera: N = 3; mice receiving immune BAFFR-/- sera: N = 6) and statistics were performed using a log rank test for significance. G, BAFFR-/- mice were vaccinated with WNVE-αCD180 and infected with WNV (as in A), 6 months later bone marrows were harvested to measure long-lived WNV E-specific ASCs by ELISPOT. In G, data are represented as means ± SEM from one experiment and statistics were determined by two-tailed Student’s t test. For all statistical analyses significant p values are indicated as follows: * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.