Establishment of a Cynomolgus Macaque Model for Human Adenovirus Type 55–Induced Respiratory Disease

Figures

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  Figure 1.. Study design and monitoring parameters after PBS administration and HAdV-55 infection in cynomolgus macaques. Cynomolgus macaques (n = 3) were initially administered PBS simultaneously via IT and IN routes. Following a 20-day recovery period, the macaques were infected with HAdV-55 using the same administration routes as PBS and monitored using the same observational criteria. Body weight, respiration rate, body temperature, oxygen saturation, heart rate, and food consumption were monitored from day 0 to day 14 after injection PBS and HAdV-55 infection.

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  Figure 2.. Physiological monitoring following PBS administration and HAdV-55 infection. A, Physiological parameters, including body weight (upper left), respiratory rate (upper middle), body temperature (upper right), oxygen saturation (SpO2) (lower left), and heart rate (lower middle), were monitored after PBS administration or HAdV-55 infection. B, Food consumption post PBS administration or HAdV-55 infection, were monitored. The gray areas in the graphs indicate the normal ranges for each parameter. C, Clinical signs were monitored daily for 14 d following infection. A total clinical score was calculated for each animal as the sum of seven parameters: general appearance (0–15), skin and fur (0–10), nose/mouth/eyes and head (0–5), respiration (0–15), feces and urine (0–10), food intake (0–10), and activity (0–35). To enhance visualization of actual scoring range, the y-axis maximum for the bar graph was set to 15 rather than the default value of 100. Data presented as mean ± SD. Statistical significances between PBS-injected and HAdV-55-infected groups at each time point were assessed using the Kruskal–Wallis test (n = 3 per group; *P < .05).

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  Figure 3.. Lung pathology monitoring via CT scans before and after PBS administration and HAdV-55 infection. Lung pathological changes were assessed by CT scans before and 14 d after PBS administration (A) or HAdV-55 infection (B). C, Cross-sectional CT images highlight peri-bronchial consolidation (circles) and ground-glass opacity (arrows).

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  Figure 4.. Histological analysis of lung tissue in cynomolgus macaques 14 d post-HAdV-55 infection. A, Granulomatous inflammation (scale bar: 100 μm), (B) inflammatory cell infiltration (scale bar: 50 μm), and (C) increased macrophage presence (scale bar: 50 μm) were observed. Arrowheads indicate the infiltration of inflammatory cells (arrows, B) and macrophages (arrows, C). D, The lung pathology was observed in H&E-stained sections and histology score in each animal was determined as described in Materials and Methods.

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  Figure 5.. Viral titers in cynomolgus macaques post PBS administration or HAdV-55 infection. Swabs from the nose and mouth, as well as blood samples, were collected from cynomolgus macaques following PBS administration and HAdV-55 infection. Viral titers were measured in blood (A), nasal swab (B), and oral swab (C) using RT-PCR (upper panel) and plaque assay (lower panel), respectively. Samples collected at 0 d postinjection or infection represent preinoculation baselines. Data presented as mean ± SD (n = 3 per group).

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  Figure 6.. Blood cell profile and Serum cytokine level changes following PBS administration and HAdV-55 infection in cynomolgus macaques. A, Complete blood count (CBC) analysis. Parameters included WBCs, neutrophils, lymphocytes, monocytes, eosinophils, basophils, and large unstained cells. Gray shaded areas represent the normal reference ranges for each parameter. B, Serum cytokine levels, TNF-α, IFN-α, IFN-β, IFN-γ, IL-6, IL-8, IL-1β, IL-4, and IL-10 were monitored using ELISA. Data presented as mean ± SD. Comparisons between PBS-injected and HAdV-55–infected groups at each time point were performed using the Kruskal–Wallis test (*P < .05; n = 3 per group).

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  Figure 7.. Humoral and cellular immune responses following PBS administration and HAdV-55 infection. Binding antibody titers (A) were measured using ELISA, neutralizing antibody titers (B) using PRNT, and IFN-γ-producing cellular immune responses (C) using ELISPOT assay. Data presented as mean ± SD. Kruskal–Wallis test was used to compare PBS-injected and HAdV-55 groups at each time point (*P < .05; n = 3).