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Tytuł:
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Ex vivo model of herpes simplex virus type I dendritic and geographic keratitis using a corneal active storage machine.
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Autorzy:
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Courrier E; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.
Maurin C; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.
Lambert V; Department of Ophthalmology, University Hospital, Saint-Etienne, France.
Renault D; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.
Bourlet T; Laboratory of Infectious Agents and Hygiene GIMAP-EA3064, University Hospital & University Jean Monnet, Saint-Etienne, France.
Pillet S; Laboratory of Infectious Agents and Hygiene GIMAP-EA3064, University Hospital & University Jean Monnet, Saint-Etienne, France.
Verhoeven PO; Laboratory of Infectious Agents and Hygiene GIMAP-EA3064, University Hospital & University Jean Monnet, Saint-Etienne, France.
Forest F; Department of Pathology, University Hospital, Saint-Etienne, France.
Perrache C; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.
He Z; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.
Garcin T; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.; Department of Ophthalmology, University Hospital, Saint-Etienne, France.
Rousseau A; Department of Ophthalmology, Bicêtre Hospital, APHP, South Paris University, Le Kremlin-Bicêtre, France.; Center for Immunology of Viral Infections and Autoimmune Diseases, IMVA, UMR, INSERM, CEA, South Paris University, Fontenay-aux-Roses, France.
Labetoulle M; Department of Ophthalmology, Bicêtre Hospital, APHP, South Paris University, Le Kremlin-Bicêtre, France.; Center for Immunology of Viral Infections and Autoimmune Diseases, IMVA, UMR, INSERM, CEA, South Paris University, Fontenay-aux-Roses, France.
Gain P; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.; Department of Ophthalmology, University Hospital, Saint-Etienne, France.
Thuret G; Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.; Department of Ophthalmology, University Hospital, Saint-Etienne, France.; Institut Universitaire de France, Paris, France.
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Źródło:
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PloS one [PLoS One] 2020 Jul 22; Vol. 15 (7), pp. e0236183. Date of Electronic Publication: 2020 Jul 22 (Print Publication: 2020).
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Original Publication: San Francisco, CA : Public Library of Science
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MeSH Terms:
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Cornea/*pathology
Herpesvirus 1, Human/*pathogenicity
Keratitis, Herpetic/*pathology
Organ Culture Techniques/*instrumentation
Organ Preservation/*instrumentation
Aged ; Aged, 80 and over ; Cornea/diagnostic imaging ; Cornea/ultrastructure ; Cornea/virology ; Host Microbial Interactions ; Humans ; Keratitis, Herpetic/diagnosis ; Keratitis, Herpetic/drug therapy ; Keratitis, Herpetic/virology ; Microscopy, Electron, Transmission ; Middle Aged ; Organ Culture Techniques/methods ; Organ Preservation/methods ; Slit Lamp Microscopy
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Entry Date(s):
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Date Created: 20200723 Date Completed: 20200930 Latest Revision: 20240329
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Update Code:
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20240329
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PubMed Central ID:
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PMC7375596
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DOI:
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10.1371/journal.pone.0236183
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PMID:
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32697805
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Background: Herpetic keratitis (HK) models using whole human corneas are essential for studying virus-host relationships, because of high species specificity and the role of interactions between corneal cell populations that cell culture cannot reproduce. Nevertheless, the two current corneal storage methods (hypothermia and organ culture (OC)) do not preserve corneas in good physiological condition, as they are characterized by epithelial abrasion, stromal oedema, and excessive endothelial mortality.
Methods: To rehabilitate human corneas intended for scientific use, we used an active storage machine (ASM) that restores two physiological parameters that are essential for corneal homeostasis: intraocular pressure and storage medium renewal (21mmHg and 2.6 μL/min, respectively). ASM storage regenerates a normal multilayer epithelium in 2 weeks. We infected six pairs of corneas unsuitable for graft by inoculating the epithelium with herpes simplex virus type 1 (HSV-1), and compared each ASM-stored cornea with the other cornea stored in the same medium using the conventional OC method.
Results: Only corneas in the ASM developed a dendritic (n = 3) or geographic (n = 2) epithelial ulcer reproducing typical HSV-1-induced clinical lesions. Corneas in OC showed only extensive desquamations. None of the uninfected controls showed epithelial damage. Histology, immunohistochemistry, transmission electron microscopy and polymerase chain reaction on corneal tissue confirmed infection in all cases (excluding negative controls).
Conclusions: The ASM provides an innovative ex vivo model of HK in whole human cornea that reproduces typical epithelial lesions.
Competing Interests: Please note that PG and GT are inventors of the active storage machine on “patent US 20160029618A1” submitted by Jean Monnet University that covers “Medical device intended for long-term storage of a cornea, or for ex vivo experimentation on a human or animal cornea”. The other authors have no proprietary or commercial interest in any materials discussed in this article."I can confirm that this does not alter our adherence to PLOS ONE policies on sharing data and materials.
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