title: Ultrashort pulsed laser treatment inactivates viruses by inhibiting viral replication and transcription in the host nucleus.
abstract: Ultrashort pulsed laser irradiation is a new method for virus reduction in pharmaceuticals and blood products. Current evidence suggests that ultrashort pulsed laser irradiation inactivates viruses through an impulsive stimulated Raman scattering process, resulting in aggregation of viral capsid proteins. However, the specific functional defect(s) in viruses inactivated in this manner have not been demonstrated. This information is critical for the optimization and the extension of this treatment platform to other applications. Toward this goal, we investigated whether viral internalization, replication, or gene expression in cells were altered by ultrashort pulsed laser irradiation. Murine Cytomegalovirus (MCMV), an enveloped DNA virus, was used as a model virus. Using electron and fluorescence microscopy, we found that laser-treated MCMV virions successfully internalized in cells, as evidenced by the detection of intracellular virions, which was confirmed by the detection of intracellular viral DNA via PCR. Although the viral DNA itself remained polymerase-amplifiable after laser treatment, no viral replication or gene expression was observed in cells infected with laser-treated virus. These results, along with evidence from previous studies, support a model whereby the laser treatment stabilizes the capsid, which inhibits capsid uncoating within cells. By targeting the mechanical properties of viral capsids, ultrashort pulsed laser treatment represents a unique potential strategy to overcome viral mutational escape, with implications for combatting emerging or drug-resistant pathogens.
authors.affiliation: Department of Radiology, Washington University School of Medicine, St Louis, MO 63110, United States. Electronic address: tsens@wusm.wustl.edu.
authors.forename: Shaw-Wei D
authors.initials: SW
authors.lastname: Tsen
authors.affiliation: Molecular Microbiology, Washington University School of Medicine, St Louis, MO 63110, United States. Electronic address: tjchapa@wustl.edu.
authors.forename: Travis
authors.initials: T
authors.lastname: Chapa
authors.affiliation: Molecular Microbiology, Washington University School of Medicine, St Louis, MO 63110, United States. Electronic address: beatty@borcim.wustl.edu.
authors.forename: Wandy
authors.initials: W
authors.lastname: Beatty
authors.affiliation: Department of Radiology, Washington University School of Medicine, St Louis, MO 63110, United States. Electronic address: baogangx@mir.wustl.edu.
authors.forename: Baogang
authors.initials: B
authors.lastname: Xu
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, United States; Center for Biophysics, Arizona State University, Tempe, AZ 85287, United States. Electronic address: tsen@asu.edu.
authors.forename: Kong-Thon
authors.initials: KT
authors.lastname: Tsen
authors.affiliation: Department of Radiology, Washington University School of Medicine, St Louis, MO 63110, United States; Biochemistry and Molecular Biophysics, Washington University School of Medicine, St Louis, MO 63110, United States; Biomedical Engineering, Washington University School of Medicine, St Louis, MO 63110, United States. Electronic address: achilefus@mir.wustl.edu.
authors.forename: Samuel
authors.initials: S
authors.lastname: Achilefu
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins / Q000378:metabolism / Q000528:radiation effects; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D015870:Gene Expression / Q000528:radiation effects; D028022:Low-Level Light Therapy; D051379:Mice; D008807:Mice, Inbred BALB C; D018146:Muromegalovirus / Q000528:radiation effects*; D066329:Protein Aggregates / Q000528:radiation effects*; D014158:Transcription, Genetic / Q000528:radiation effects; D038301:Virus Inactivation / Q000528:radiation effects*; D053586:Virus Internalization / Q000528:radiation effects; D014779:Virus Replication / Q000528:radiation effects*
chemical_list: D036022:Capsid Proteins; D004279:DNA, Viral; D066329:Protein Aggregates
keywords: Murine cytomegalovirus; Pathogen inactivation; Pathogen reduction; Ultrashort pulsed lasers
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