In vitro exposure system for study of aerosolized influenza virus

Creager, Hannah M.; Zeng, Hui; Pulit-Penaloza, Joanna A.; Maines, Taronna R.; Tumpey, Terrence M.; Belser, Jessica A.

doi:10.1016/j.virol.2016.10.007

Cited by 15 publications

(10 citation statements)

References 49 publications

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“…At 33°C, ID 50 s were similar, except in the case of the NL/230 virus, which had an ID 50 of 83 PFU at 33°C, compared to 10 PFU at 37°C. Consistent with data for respiratory cells (18), temperature had little effect on the growth of the seasonal virus Pan/99, whereas the growth of the avian viruses was attenuated at the lower temperature. The effects of temperature on the curve shape were similar across inoculation doses, but differences in peak titers were greater at lower exposure doses, possibly because the low dosage delayed replication at both temperatures such that titers had not plateaued by 96 h postinfection.…”

Section: Resultssupporting

confidence: 84%

“…Although ocular aerosol challenge has been used in animals, to our knowledge, this inoculation method has not been used with human ocular cells or tissues. We previously found that primary human alveolar epithelial cells were less susceptible to infection with certain viruses when inoculated via aerosol than when inoculated by more traditional methods (18). To determine whether a similar phenomenon might explain why both seasonal and avian respiratory viruses, previously shown to productively infect multiple ocular cell types (8,9,11,12,23), rarely establish ocular infection in humans, we inoculated corneal epithelial cells by the aerosol route.…”

Section: Discussionmentioning

confidence: 99%

“…MOIs for corneal constructs were calculated based on the midpoint (75,000 cells) of the manufacturer's estimate of 50,000 to 100,000 cells on the apical surface (MatTek). Aerosol inoculations were performed as described previously (18). Briefly, after the removal of apical medium, if present, cells were placed into an aerosol exposure chamber for a 15-min exposure to aerosolized virus, followed by 5 min of room air to purge viral aerosols from the system.…”

Section: Methodsmentioning

confidence: 99%

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Infection and Replication of Influenza Virus at the Ocular Surface

Creager

Kumar

Zeng

et al. 2018

J Virol

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Although influenza viruses typically cause respiratory tract disease, some viruses, particularly those with an H7 hemagglutinin, have been isolated from the eyes of conjunctivitis cases. Previous work has shown that isolates of multiple subtypes from both ocular and respiratory infections are capable of replication in human ocular tissues and corneal or conjunctival cell monolayers, leaving the determinants of ocular tropism unclear. Here, we evaluated the effect of several variables on tropism for ocular cells cultured and examined the potential effect of the tear film on viral infectivity. All viruses tested were able to replicate in primary human corneal epithelial cell monolayers subjected to aerosol inoculation. The temperature at which cells were cultured postinoculation minimally affected infectivity. Replication efficiency, in contrast, was reduced at 33°C relative to that at 37°C, and this effect was slightly greater for the conjunctivitis isolates than for the respiratory ones. With the exception of a seasonal H3N2 virus, the subset of viruses studied in multilayer corneal tissue constructs also replicated productively after either aerosol or liquid inoculation. Human tears significantly inhibited the hemagglutination of both ocular and nonocular isolates, but the effect on viral infectivity was more variable, with tears reducing the infectivity of nonocular isolates more than ocular isolates. These data suggest that most influenza viruses may be capable of establishing infection if they reach the surface of ocular cells but that this is more likely for ocular-tropic viruses, as they are better able to maintain their infectivity during passage through the tear film. The potential spread of zoonotic influenza viruses to humans represents an important threat to public health. Unfortunately, despite the importance of cellular and tissue tropism to pathogenesis, determinants of influenza virus tropism have yet to be fully elucidated. Here, we sought to identify factors that limit the ability of most influenza viruses to cause ocular infection. Although ocular symptoms in humans caused by avian influenza viruses tend to be relatively mild, these infections are concerning due to the potential of the ocular surface to serve as a portal of entry for viruses that go on to establish respiratory infections. Furthermore, a better understanding of the factors that influence infection and replication in this noncanonical site may point toward novel determinants of tropism in the respiratory tract.

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Section: Resultssupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Infection and Replication of Influenza Virus at the Ocular Surface

Creager

Kumar

Zeng

et al. 2018

J Virol

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“…Ferrets have represented a critical in vivo model for these activities, and it is likely that the role this species plays in our understanding of influenza virus pathogenicity, transmissibility, and tropism will only increase in the coming years. Advances in cell culture models have sought to more closely emulate the complexity of in vivo models; examples include the development of physiologically relevant inoculation routes and the generation of organoid tissues that are representative of the human respiratory tract (Creager et al 2017;Hui et al 2018). Similarly, ex vivo primary differentiated cultures of ferret tissue-specific cells provide an additional link between in vivo and in vitro experimentation, supporting and enhancing studies investigating viral pathogenicity and tropism (Zeng et al 2013(Zeng et al , 2019.…”

Section: Discussionmentioning

confidence: 99%

Ferreting Out Influenza Virus Pathogenicity and Transmissibility: Past and Future Risk Assessments in the Ferret Model

Belser

Pulit-Penaloza

Maines

2019

Cold Spring Harb Perspect Med

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“…Here, H1N1‐containing aerosols with a mass median aerodynamic diameter of 1.27 ± 0.61 μm were generated by an animal nose‐only aerosol exposure device. In addition, some comparative studies have demonstrated that compared with droplet instillation, inhalation of H5N1‐ and H7N9‐containing aerosols represent a more natural route of human exposure . Therefore, the aerosol inhalation method, which directly delivers H1N1‐containing aerosols to the noses of exposed BALB/C mice via a nose‐only aerosol exposure device, was selected to mimic the natural route of human infection with influenza A(H1N1).…”

Section: Introductionmentioning

confidence: 99%

Establishment of BALB/C mouse models of influenza A H1N1 aerosol inhalation

Hao

et al. 2019

Journal of Medical Virology

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Influenza A (H1N1) is a rapidly spreading acute respiratory illness that remains a worldwide burden on public health. To simulate natural infection routes, BALB/C mice were challenged with the H1N1 virus by aerosol and intranasal instillation routes. We compared the weight change and survival of the mice for 14 consecutive days after infection. The infected mice were euthanized at days 3, 5, 7, and 9 to perform necropsies, lung pathological analyses, viral titers measurement, and lung cytokines examination. The aerosol-treated mice showed clinical symptoms on day 4, obvious lung lesions on day 5, rapid weight loss on day 7, peak virus replication in the lungs on days 7 to 9, and bronchial epithelial hyperplasia on day 9. However, after intranasal instillation, the mice exhibited clinical signs on day 2, rapid weight loss and obvious lung lesions on day 3, and peak virus replication in the lungs on days 3 to 5; no bronchial epithelial hyperplasia was detected. High levels of proinflammatory cytokines and chemokines were detected in the lungs of infected mice by both two routes.Disease and lung lesion progressions were slower in the mice that inhaled H1N1containing aerosols than in those treated by intranasal instillation, and lung lesions were homogeneous in the aerosol group and heterogeneous in the intranasal group. In this study, BALB/C mouse models of H1N1 virus aerosol inhalation were successfully established and compared with mouse models of intranasal inoculation, aerosol mouse models had an infection route and lung pathology characteristics that more closely resembled those observed in humans. K E Y W O R D Saerosol inhalation, H1N1 virus aerosol, intranasal instillation, mouse models

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In vitro exposure system for study of aerosolized influenza virus

Cited by 15 publications

References 49 publications

Infection and Replication of Influenza Virus at the Ocular Surface

Infection and Replication of Influenza Virus at the Ocular Surface

Ferreting Out Influenza Virus Pathogenicity and Transmissibility: Past and Future Risk Assessments in the Ferret Model

Establishment of BALB/C mouse models of influenza A H1N1 aerosol inhalation

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