Article Figures & Data
Figures
- FIGURE 1.
The experimental protocol of this study.
Numbers of mice in each group are shown in the inset table. Small-scale preliminary experiments were performed to determine the conditions of this study.
- FIGURE 2.
ARNAX adjuvant shows some advantages compared with Alum in vaccination.
Adjuvant efficacy and lung histopathology in mice immunized with recombinant S protein with ARNAX120. Female BALB/c mice were vaccinated with each set of Ag/adjuvant. Mice immunized with 0.1 μg S protein with or without adjuvant were challenged with 106 TCID50 of mouse-adapted SARS‐CoV (n = 6–10). (A) Serum neutralizing Ab titers after the second immunization. The line indicates the limit of detection (<4). *p < 0.05; ***p < 0.001, via Dunn's multiple comparisons test following the Kruskal–Wallis test. (B) Body weight changes after SARS-CoV challenge infection. (C) Survival curves after SARS‐CoV challenge infection. Comparisons of survival with respect to the control group were performed using the log-rank test followed by Kaplan–Meier survival analysis. (D) Virus titers in lungs on day 3 postchallenge. **p < 0.01, via Dunn's multiple comparisons test following the Kruskal–Wallis test. (E) Lung histopathological findings on day 10 postchallenge. Upper panels, low magnification (scale bars, 200 µm); middle panels, high magnification (scale bars, 100 µm); lower panels, high magnification of the lung histopathological findings from the mice with the eosinophil infiltration were detected via eosinophil staining using C.E.M. Stain Kit (scale bars, 20 µm). The red arrows indicate representative eosinophils, and the blue arrows indicate plasma cells. Results of the PBS pretreated controls on day 5 postchallenge. Each image represents many fields from a specimen of an individual animal. (F) Number of eosinophils per lung section (n = 3–6) on day 10 postchallenge. Five 147,000-μm2 regions around the pulmonary bronchiole of each mouse were counted.
- FIGURE 3.
Histopathology of alveolar area of the lung in mice treated with Ag + adjuvant.
(A) The alveolar regions in the lung specimens (Fig. 2) were enlarged. The different fields were shown with a high magnification under the microscope. Score of leukocyte infiltration was given in each panel as in Iwata-Yoshikawa et al. (14). (B) Representative images of alveoli with eosinophils (arrowheads or asterisks) from Ag + adjuvant-treated mice. Each image represents many fields from a specimen of an individual animal. The insets are higher-magnification images of the boxed area. H&E staining. Scale bars, 50 µm; 20 µm (insets).
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