Lack of iNOS facilitates MCMV spread in the retina

Ming Zhang, Jun Zhou, Brendan Marshall, Hua Xin, Sally S. Atherton

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

PURPOSE. The purposes of this study were to identify iNOS-producing retinal cells and to determine whether lack of iNOS facilitates MCMV spread and replication in the retina. METHODS. Immunosuppressed (IS) iNOS-/- mice or C57BL/6 (wild-type) mice were inoculated with 5 × 104 PFU of MCMV K181 strain (K181) via the supraciliary route. Injected eyes were collected at several times after inoculation and examined by plaque assay for replicating virus, RT-PCR for iNOS RNA, Western blot for iNOS protein and by staining for MCMV early antigen (EA), iNOS, and retinal cell antigens. RESULTS. iNOS mRNA and iNOS proteins were expressed in the MCMV-injected eye of wild-type mice. Most iNOS-producing cells were F4/80-positive, including macrophages, RPE-derived macrophages, and resident microglia. Significantly higher titers of virus were recovered from the injected eyes, and more infected cells were detected in the retina of IS iNOS-/- mice than in IS wild-type mice. Retinal necrosis and loss of retinal architecture throughout the retina were noted in IS iNOS-/- mice, whereas cytomegalic cells and retinitis were present only in the peripheral retina of IS wild-type mice. CONCLUSIONS. iNOS produced by macrophages, especially resident macrophages including microglia and RPE derived macrophages, plays an important role in limiting spread of MCMV in the retina.

Original languageEnglish (US)
Pages (from-to)285-292
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume48
Issue number1
DOIs
StatePublished - Jan 1 2007

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Retina
Macrophages
Microglia
Retinitis
Antigens
Viral Load
Inbred C57BL Mouse
Proteins
Necrosis
Western Blotting
RNA
Staining and Labeling
Viruses
Polymerase Chain Reaction
Messenger RNA

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Lack of iNOS facilitates MCMV spread in the retina. / Zhang, Ming; Zhou, Jun; Marshall, Brendan; Xin, Hua; Atherton, Sally S.

In: Investigative Ophthalmology and Visual Science, Vol. 48, No. 1, 01.01.2007, p. 285-292.

Research output: Contribution to journalArticle

Zhang, Ming ; Zhou, Jun ; Marshall, Brendan ; Xin, Hua ; Atherton, Sally S. / Lack of iNOS facilitates MCMV spread in the retina. In: Investigative Ophthalmology and Visual Science. 2007 ; Vol. 48, No. 1. pp. 285-292.
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N2 - PURPOSE. The purposes of this study were to identify iNOS-producing retinal cells and to determine whether lack of iNOS facilitates MCMV spread and replication in the retina. METHODS. Immunosuppressed (IS) iNOS-/- mice or C57BL/6 (wild-type) mice were inoculated with 5 × 104 PFU of MCMV K181 strain (K181) via the supraciliary route. Injected eyes were collected at several times after inoculation and examined by plaque assay for replicating virus, RT-PCR for iNOS RNA, Western blot for iNOS protein and by staining for MCMV early antigen (EA), iNOS, and retinal cell antigens. RESULTS. iNOS mRNA and iNOS proteins were expressed in the MCMV-injected eye of wild-type mice. Most iNOS-producing cells were F4/80-positive, including macrophages, RPE-derived macrophages, and resident microglia. Significantly higher titers of virus were recovered from the injected eyes, and more infected cells were detected in the retina of IS iNOS-/- mice than in IS wild-type mice. Retinal necrosis and loss of retinal architecture throughout the retina were noted in IS iNOS-/- mice, whereas cytomegalic cells and retinitis were present only in the peripheral retina of IS wild-type mice. CONCLUSIONS. iNOS produced by macrophages, especially resident macrophages including microglia and RPE derived macrophages, plays an important role in limiting spread of MCMV in the retina.

AB - PURPOSE. The purposes of this study were to identify iNOS-producing retinal cells and to determine whether lack of iNOS facilitates MCMV spread and replication in the retina. METHODS. Immunosuppressed (IS) iNOS-/- mice or C57BL/6 (wild-type) mice were inoculated with 5 × 104 PFU of MCMV K181 strain (K181) via the supraciliary route. Injected eyes were collected at several times after inoculation and examined by plaque assay for replicating virus, RT-PCR for iNOS RNA, Western blot for iNOS protein and by staining for MCMV early antigen (EA), iNOS, and retinal cell antigens. RESULTS. iNOS mRNA and iNOS proteins were expressed in the MCMV-injected eye of wild-type mice. Most iNOS-producing cells were F4/80-positive, including macrophages, RPE-derived macrophages, and resident microglia. Significantly higher titers of virus were recovered from the injected eyes, and more infected cells were detected in the retina of IS iNOS-/- mice than in IS wild-type mice. Retinal necrosis and loss of retinal architecture throughout the retina were noted in IS iNOS-/- mice, whereas cytomegalic cells and retinitis were present only in the peripheral retina of IS wild-type mice. CONCLUSIONS. iNOS produced by macrophages, especially resident macrophages including microglia and RPE derived macrophages, plays an important role in limiting spread of MCMV in the retina.

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