Fig. 3: Generation and characterization of second-generation MV-based candidate ZIKV vaccines. | npj Vaccines

Fig. 3: Generation and characterization of second-generation MV-based candidate ZIKV vaccines.

From: Measles-based Zika vaccine induces long-term immunity and requires NS1 antibodies to protect the female reproductive tract

Fig. 3

a Second-generation vaccine constructs. b Immunofluorescence staining of Vero cells were infected at MOI-0.1 for 72 h with the recovered modified MV-ZIKV candidate vaccines and control viruses. The permeabilized cells were stained for MV using an α-MV N protein (Sigma, NP-Cl25) mouse monoclonal conjugated with dylight488 (green color changed to Yellow for visualization), for the ZIKV-E using a mouse monoclonal antibody (Biofront-1176-56) and for ZIKV NS1 using human monoclonal antibody EB9. ZIKV-E is stained with a secondary α-mouse AF568 (red color changed to Cyan for visualization) antibody and ZIKV-NS1 is stained with a secondary goat α-human AF-647 (far-red color changed to Magenta for visualization) antibody. Confocal images were taken using NIKON-A1R, 60X magnification, 2.5X zoom. The scale bar measures 30 μm. The data is representing one replicate of n = 3. c Sucrose-purified virions were analyzed on SDS-PAGE (10%) stained with SYPRO Ruby. Zika virus PRVABC59 was loaded with the control virus. The data is representing one replicate of n = 3. d Western blot analysis of sucrose purified virions probed for ZIKV-E (Biofront mouse mAb-1176) and MV-H (Rabbit polyclonal sera). Zika virus PRVABC59 was loaded as the control virus. The data is representing one replicate of n = 3. e Western blot analysis of cell lysates of Vero cells infected with MV-ZIKV candidate vaccine at MOI-5. Zika virus PRVABC59 infected cell lysates were loaded as the control. The blot was probed for ZIKV-E (Biofront mouse mAb), ZIKV-NS1 (Abcam, B4 mouse mAb) and MV-N (Sigma, NP-Cl25). The data is representing one replicate of n = 2.

Back to article page