May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Does Nicotine Treatment Effect the Downregulation of NF–B During Human Cytomegalovirus Infection of Retinal Pigment Epithelial Cells?
Author Affiliations & Notes
  • A.E. Buckner
    Jones Eye Institute, Univ Arkansas Medical Sciences, Little Rock, AR
  • J.Y. Chang
    Jones Eye Institute, Univ Arkansas Medical Sciences, Little Rock, AR
  • R.D. Dix
    Jones Eye Institute, Univ Arkansas Medical Sciences, Little Rock, AR
  • Footnotes
    Commercial Relationships  A.E. Buckner, None; J.Y. Chang, None; R.D. Dix, None.
  • Footnotes
    Support  Arkansas Tobacco Settlement Grant & RPB
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 1663. doi:
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      A.E. Buckner, J.Y. Chang, R.D. Dix; Does Nicotine Treatment Effect the Downregulation of NF–B During Human Cytomegalovirus Infection of Retinal Pigment Epithelial Cells? . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1663.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose. Human cytomegalovirus (HCMV) infection of retinal pigment epithelial (RPE) cells results in downregulation of the cellular transcription factor NF–ΚB, an observation that may explain the unusual semipermissiveness of RPE for HCMV replication. Since experimental mouse models of HCMV retinitis suggest the RPE to be an initial target for virus infection, we became interested in the possible influence of tobacco smoking as a cofactor in the development of HCMV retinal disease. We therefore performed pilot studies to test the hypothesis that nicotine counteracts the dampening effect of HCMV infection on NF–ΚB in RPE, thereby increasing the permissiveness of RPE for HCMV replication as occurs in fibroblasts in which HCMV infection enhances NF–ΚB expression. Methods.The ARPE–19 cell line (cells with properties similar to human RPE cells) was used for this study. Toxicity experiments were performed to assess ARPE–19 cells grown in different amounts of nicotine for viability. Experiments were then performed in which monolayers of ARPE–19 cells were transfected by cationic lipid–mediated NF–ΚB luciferase DNA (pNF–ΚB–Luc), inoculated with HCMV [Towne] (moi=1) at 24 hours post–transfection, and maintained in the absence or presence of a nontoxic and physiologic dose of nicotine (100nM) at 1 hr prior to or 1 hr after HCMV inoculation. All monolayers were harvested at 4 hrs postinfection, lysed using a reporter buffer, and quantified for luciferase activity as a measure of NF–ΚB activity. Results. Whereas HCMV–infected ARPE–19 cells without nicotine treatment showed a dramatic decrease in NF–ΚB levels, nicotine treatment reduced this decrease substantially, but did not abolish it completely. Nicotine treatment of uninfected ARPE–19 cells had no effect on baseline NF–ΚB levels. Conclusions. Treatment of HCMV–infected ARPE–19 cells with nicotine at a nontoxic and physiologic dose dampened the downregulation of NF–ΚB observed in HCMV–infected ARPE–19 cells without nicotine treatment. We predict that this effect on NF–ΚB expression will promote permissiveness of RPE for productive HCMV replication. This finding suggests that tobacco smoking may promote lytic replication of HCMV within RPE leading to retinal disease.

Keywords: cytomegalovirus • retinal pigment epithelium • gene/expression 
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