December 2002
Volume 43, Issue 13
ARVO Annual Meeting Abstract  |   December 2002
Mouse Cornea Transduced with an Adenoviral Construct Expressing Murine Interferon-Beta (Ad:IFN-B) Antagonizes Herpes Simplex Virus Type 1 (HSV-1)-Induced Mortality
Author Affiliations & Notes
  • K AL-Khatib
    Microbiology and Immunology Univ of Oklahoma Health Sciences Ctr Oklahoma City OK
  • WP Halford
    Microbiology and Immunology Tulane University Medical Center New Orleans LA
  • DJ J Carr
    Department of Ophthalmology University of Oklahoma Health Sciences Center Oklahoma City OK
  • Footnotes
    Commercial Relationships   K. AL-Khatib, None; W.P. Halford, None; D.J.J. Carr, None. Grant Identification: NIH Grant EY12409
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 4333. doi:
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      K AL-Khatib, WP Halford, DJ J Carr; Mouse Cornea Transduced with an Adenoviral Construct Expressing Murine Interferon-Beta (Ad:IFN-B) Antagonizes Herpes Simplex Virus Type 1 (HSV-1)-Induced Mortality . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4333.

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

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Abstract: : Purpose:To determine the success of in situ transduction and anti-viral efficacy of the Ad:IFN-b construct against ocular HSV-1 infection. Methods:The Ad:IFN-b construct was generated by ligation of the murine IFN-b coding sequence into Sal I and Xba I sites of the adenovirus shuttle vector pBHad-TRE and co-transfecting 293 cells with the shuttle vector and a replication deficient adenoviral vector (D1-D3) containing the green fluorescent protein (Ad:GFP). The recombinant adenovirus was selected by the loss of GFP expression, plaque purified three times, and characterized by Southern blot and PCR for the presence of the murine-IFN-b transgene. The Ad:IFN-b and reporter (Ad:GFP) constructs were used to transduce mouse cornea and determine longevity of expression as well as induction of IFN-responsive genes (OAS and PKR) and production of biologically active IFN-b. Survival studies in recipient mice infected with HSV-1 were also conducted. Results:GFP expression was observed within 48 h post transduction (PT) and continued for up to 14 days PT. Biologically active IFN-b (2.5 U/button) was detected in 3 out of 5 corneas transduced. No IFN was detected in control transduced (Ad:Null) corneas. OAS but not PKR expression was found to be enhanced in the cornea of Ad:IFN-b recipients compared to control (Ad:Null)-transduced cornea. Mice transduced with the Ad:IFN-b contruct 48 – 96 h prior to ocular infection with HSV-1 (McKrae strain, 150 pfu/eye) showed enhanced (p<.05) survival (67-80%, n=15/group) compared to Ad:Null transduced mice (20-30% survival, n=15). Conclusion:The adenovirus vector is an efficient approach to deliver the murine IFN-b transgene to the eye of mice in terms of expression, induction of IFN-responsive genes, and antagonizing HSV-1-mediated mortality.

Keywords: 425 herpes simplex virus • 419 gene transfer/gene therapy • 380 cytokines/chemokines 

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