A novel orally infected hamster model for Coxsackievirus A16 hand-foot-and-mouth disease and encephalomyelitis

Abstract

Coxsackievirus A16 (CV-A16) is one of the major causes of mild and self-limiting hand-foot-and-mouth disease (HFMD) in young children, which may occasionally leads to serious neurological complications. In this study, we had developed a novel, consistent, orally infected CV-A16 HFMD hamster model with encephalomyelitis. Four groups of 7-day-old hamsters in a kinetic study were orally infected with mouse-adapted CV-A16 strains and sacrificed at 1–4 days post infection (dpi), respectively. Tissues were studied by light microscopy, immunohistochemistry to detect viral antigens, in situ hybridization to detect viral RNA, and by viral titration. In a separate transmission experiment, orally infected index hamsters were housed together with contact hamsters to investigate oral and fecal viral shedding by virus culture and reverse transcription polymerase chain reaction (RT-PCR). At severe infection/death endpoints, index and contact hamster infection were also histopathologically analyzed. In the kinetic study, infected hamsters developed signs of infection at 4 dpi. Viral antigens/RNA were localized to brainstem (medulla/pons; reticular formation and motor trigeminal nucleus) and spinal cord anterior horn neurons, oral squamous epithelia and epidermis from 3 to 4 dpi. Salivary and lacrimal glands, myocardium, brown adipose tissue, intestinal smooth muscle, and skeletal muscle infection was also demonstrated. Viremia at 1 dpi and increasing viral titers in various tissues were observed from 2 dpi. In the transmission study, all contact hamsters developed disease 3–5 days later than index hamsters, but demonstrated similar histopathological findings at endpoint. Viral culture and RT-PCR positive oral washes and feces confirmed viral shedding. Our hamster model, orally infected by the natural route for human infection, confirmed CV-A16 neurotropism and demonstrated squamous epitheliotropism reminiscent of HFMD, attributes not found in other animal models. It should be useful to investigate neuropathogenesis, model person-to-person transmission, and for testing antiviral drugs and vaccines.

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Fig. 1: IHC detection of viral antigens in CV-A16-infected hamsters (kinetic study).
Fig. 2: Approximate CNS distribution of viral antigen-positive neurons in CV-A16-infected hamsters.
Fig. 3: Mean viral titers in various tissues derived from CV-A16 infected hamsters from 1 to 4 dpi groups (Kinetic study).
Fig. 4: IHC and ISH detection in a CV-A16-infected index hamster at 5 dpi.
Fig. 5: IHC and ISH detection in index and contact hamsters.

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Acknowledgements

This study was supported by the High Impact Research Grant (H20001-E00004), Fundamental Research Grant Scheme (FP038/2015A) from the Ministry of Higher Education, Malaysia Government, and Postgraduate Research Grant (PG159-2015A) from University of Malaya.

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Correspondence to Kum Thong Wong.

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Hooi, Y.T., Ong, K.C., Tan, S.H. et al. A novel orally infected hamster model for Coxsackievirus A16 hand-foot-and-mouth disease and encephalomyelitis. Lab Invest (2020). https://doi.org/10.1038/s41374-020-0456-x

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