The use of human cornea organotypic cultures to study herpes simplex virus type 1 (HSV-1)-induced inflammation

Peter Drevets, Ana Chucair-Elliott, Priyadarsini Shrestha, Jeremy Jinkins, Dimitrios Karamichos, Daniel J.J. Carr

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Purpose: To determine the utility of human organotypic cornea cultures as a model to study herpes simplex virus type 1 (HSV-1)-induced inflammation and neovascularization. Methods: Human organotypic cornea cultures were established from corneas with an intact limbus that were retrieved from donated whole globes. One cornea culture was infected with HSV-1 (104 plaque-forming units), while the other cornea from the same donor was mock-infected. Supernatants were collected at intervals post-culture with and without infection to determine viral titer (by plaque assay) and pro-angiogenic and proinflammatory cytokine concentration by suspension array analysis. In some experiments, the cultured corneas were collected and evaluated for HSV-1 antigens by immunohistochemical means. Another set of experiments measured susceptibility of human three-dimensional cornea fibroblast constructs, in the presence and absence of TGF-β1, to HSV-1 infection in terms of viral replication and the inflammatory response to infection as a comparison to the organotypic cornea cultures. Results: Organotypic cornea cultures and three-dimensional fibroblast constructs exhibited varying degrees of susceptibility to HSV-1. Fibroblast constructs were more susceptible to infection in terms of infectious virus recovered in a shorter period of time. There were changes in the levels of select pro-angiogenic or proinflammatory cytokines that were dictated as much by the cultures producing them as by whether they were infected with HSV-1 or treated with TGF-β1. Conclusion: Organotypic cornea and three-dimensional fibroblast cultures are likely useful for the identification and short-term study of novel antiviral compounds and virus replication, but are limited in the study of the local immune response to infection.

Original languageEnglish
Pages (from-to)1721-1728
Number of pages8
JournalGraefe's Archive for Clinical and Experimental Ophthalmology
Volume253
Issue number10
DOIs
StatePublished - 22 Oct 2015

Keywords

  • Cornea
  • Cytokines
  • HSV-1
  • Inflammation
  • Pro-angiogenic factors

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