yezv-n peptide (pos: 165–182) immunized rabbit serum antibody  (Cosmo Bio USA)

Journal: PLOS Pathogens

Article Title: Establishment of a lethal mouse model of emerging tick-borne orthonairovirus infections

doi: 10.1371/journal.ppat.1012101

Figure Lengend Snippet: AG129 mice inoculated intraperitoneally with 10 4 FFU of YEZV were sacrificed at 2, 4, and 6 dpi to collect organ samples. Mock-infected mice sacrificed at 6 dpi are shown as controls. Mouse organs were fixed in 10% phosphate-buffered formalin and (A) stained with hematoxylin-eosin (H&E) or (B and C) immunohistochemically stained for YEZV N. (A) Representative gross images and H&E-stained tissue images of the liver and spleen are shown. Each side of a square in the gross images is 5 mm. (B) Representative IHC images of the liver and spleen stained for YEZV N are shown with viral antigens in brown. (A and B) The scale bars on the histological images of the liver and spleen are 100 μm and 200 μm, respectively. (C) YEZV N protein (blue) and macrophage marker, Iba1 (brown) were stained by IHC double staining in the liver and spleen of YEZV-infected mice. The scale bars on IHC images of the liver and spleen tissues are 50 μm and 20 μm, respectively. Arrows indicate a cell in which both YEZV antigen and Iba1 were detected.

Article Snippet: IHC was performed using YEZV-N peptide (POS: 165–182) immunized rabbit serum (1:20,000) (Cosmo Bio, Tokyo, Japan) or anti Iba1 goat polyclonal antibody (1:500, #001–27991, Wako, Osaka, Japan) as primary antibodies.

Techniques: Infection, Staining, Marker, Double Staining Immunohistochemistry

Journal: PLOS Pathogens

Article Title: Establishment of a lethal mouse model of emerging tick-borne orthonairovirus infections

doi: 10.1371/journal.ppat.1012101

Figure Lengend Snippet: (A) The antiviral effects of T-705 in the AG129 mouse model were evaluated. AG129 mice intraperitoneally inoculated with 10 4 FFU of YEZV were orally dosed with T-705 (100 or 300 mg/kg/day) or vehicle using a stomach probe at 1 hour, 1, 2 and 3 dpi (represented by gray color on the figure). All mice were monitored for survival (left panel) and body weight changes (right panel) for 14 days. Fourteen-day survival rates and body weight changes were compared with the vehicle-treated group, 100 mg/kg/day of T-705-treated group and 300 mg/kg/day of T-705-treated group using a log-rank test. (B-E) Vehicle- or 300 mg/kg/day of T705-treated YEZV-infected AG129 mice were sacrificed at 2 and 4 dpi to collect blood and tissue samples. (B) Representative gross images and H&E-stained tissue images of the liver and spleen of mice are shown. Each side of a square in the gross images is 5 mm. The scale bars on the histological images of the liver and spleen are 100 μm and 500 μm respectively. (C) YEZV N proteins were detected by IHC in the liver and spleen of vehicle-treated and T-705-treated mice, and representative images are shown. The scale bars on the images of the liver and spleen tissues are 100 μm and 500 μm, respectively. (D) Viral titers and (E) the copy number of viral RNA (YEZV L-segment) in the liver and spleen of vehicle-treated and T-705-treated mice were quantified. Dotted lines indicated the lower limit of detection (L.O. D.) for the assay (4 FFU/mg of liver and spleen and 0.4 FFU/μL of serum). Plots lower than the L.O.D. are indicated by the half value of L.O.D. on the graph. Statistical differences in viral RNA copy number between the vehicle-treated group and T-705-group were examined by the Mann-Whitney U test, and the significant difference ( p < 0.01) is indicated as double asterisks.

Article Snippet: IHC was performed using YEZV-N peptide (POS: 165–182) immunized rabbit serum (1:20,000) (Cosmo Bio, Tokyo, Japan) or anti Iba1 goat polyclonal antibody (1:500, #001–27991, Wako, Osaka, Japan) as primary antibodies.

Techniques: Infection, Staining, MANN-WHITNEY