West Nile Virus Infection in Human and Mouse Cornea Tissue

Bradley J. Blitvich Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa.

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Tian Wang Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas.
Department of Pathology, University of Texas Medical Branch, Galveston, Texas.
Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas.

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Vandana Saxena Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas.

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Shemin Zeng Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa.
Stephen A. Wynn Institute for Vision Research, University of Iowa, Iowa City, Iowa.

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Karen M. Harmon Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa.

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Matthew D. Raymond Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa.

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Kenneth M. Goins Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa.
Iowa Lions Eye Bank, Coralville, Iowa.

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Cynthia R. Reed Iowa Lions Eye Bank, Coralville, Iowa.

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Robert F. Mullins Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa.
Stephen A. Wynn Institute for Vision Research, University of Iowa, Iowa City, Iowa.

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Mark A. Greiner Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa.
Stephen A. Wynn Institute for Vision Research, University of Iowa, Iowa City, Iowa.
Iowa Lions Eye Bank, Coralville, Iowa.

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The purpose of this study was to determine the in vitro and ex vivo susceptibility of human corneal cells to West Nile virus (WNV) infection and evaluate the ability of the virus to disseminate to the corneas of infected mice. Human corneal epithelial cells were challenged with WNV, incubated for 1–6 days, and tested for evidence of WNV infection. Viral RNA and antigen were detected at every time point, and the virus reached a peak titer of 2.5 × 107 plaque-forming units (pfu)/mL at 3 days postinoculation (PI). Corneas procured from donors were incubated in culture dishes containing WNV for 1–5 days and tested for evidence of WNV. Viral RNA and antigen were detected, and the virus reached a mean peak titer of 4.9 × 104 pfu/mL at 5 days PI. Mice were inoculated intraperitoneally with WNV, and their eyes were harvested at 2, 5, and 8 days PI and tested for evidence of WNV. Viral RNA was detected in corneas of four of nine systemically infected mice as early as 2 days PI. We conclude that human corneal cells support WNV replication in vitro and ex vivo, and WNV may disseminate into the corneas of experimentally infected mice. These findings indicate that corneal transmission cannot be ruled out as a novel mode of human-to-human WNV transmission and additional experiments should be conducted to assess this risk further.

Author Notes

* Address correspondence to Mark A. Greiner, Cornea and External Diseases, Department of Ophthalmology and Visual Sciences, University of Iowa Carver College of Medicine, 200 Hawkins Drive, Iowa City, IA 52242. E-mail: mark-greiner@uiowa.edu

Financial support: This study was supported by a grant from the Iowa Lions Eye Bank.

Authors' addresses: Bradley J. Blitvich, Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, E-mail: blitvich@iastate.edu. Tian Wang, Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, Department of Pathology, University of Texas Medical Branch, Galveston, TX, and Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, TX, E-mail: ti1wang@utmb.edu. Vandana Saxena, Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, and Department of Immunology, National AIDS Research Institute, Pune, Maharashtra, India, E-mail: vandysaxena@gmail.com. Shemin Zeng and Robert F. Mullins, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, and The Stephen A. Wynn Institute for Vision Research, University of Iowa, Iowa City, IA, E-mails: shemin-zeng@uiowa.edu and robert-mullins@uiowa.edu. Karen M. Harmon, Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, E-mail: kharmon@iastate.edu. Matthew D. Raymond, Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, and Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, E-mail: mdraymond@wisc.edu. Kenneth M. Goins, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, and Iowa Lions Eye Bank, Coralville, IA, E-mail: kenneth-goins@uiowa.edu. Cynthia R. Reed, Iowa Lions Eye Bank, Coralville, IA, E-mail: cynthia-reed@uiowa.edu. Mark A. Greiner, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IA, The Stephen A. Wynn Institute for Vision Research, University of Iowa, Iowa City, IA, and Iowa Lions Eye Bank, Coralville, IA, E-mail: mark-greiner@uiowa.edu.

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