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Analysis of cytokine mRNAs in murine herpes simplex virus type 1 retinitis
Kazuko Saitoh-Ishibashi1, Kazuki Ishibashi, Atsushi Azumi
1Division of Ophthalmology, Department of Organ Therapeutics, Kobe University Graduate School of Medicine, Kobe, Japan.
Purpose:
To investigate cytokine mRNA expression during the inflammatory process induced in the contralateral eyes by uniocular inoculation of herpes simplex virus type 1 (HSV-1) via the anterior chamber.
Methods:
BALB/c mice were inoculated in the anterior chamber with 5 x 10(4) plaque-forming units of HSV-1 (KOS). mRNA was extracted from the inflamed posterior segments of the uninoculated eyes at 0 (control), 9, 11, 14, and 21 days postinoculation (p.i.). Reverse transcription-polymerase chain reaction was performed for semiquantitative analysis of mRNA expression of interleukin (IL)-1beta, IL-2, IL-4, IL-10, IL-12p35, IL-12p40, interferon (IFN)gamma, tumor necrosis factor (TNF)alpha, transforming growth factor (TGF)beta2 and induced nitric oxide synthase (iNOS).
Results:
Peak mRNA expression of iNOS was observed at day 14 p.i. The time profiles of mRNA expression for IL-1beta, IL-2, IL-4, IL-10, IFN-gamma, TNFalpha were similar to that of iNOS, while TGFbeta2, IL-12p35, and IL-12p40 demonstrated a reverse pattern.
Conclusions:
The kinetics of the analyzed cytokines synchronized with the clinicopathological activity of the experimental murine HSV-1 retinitis. The immunosuppressive cytokines TGFbeta2 and IL-10 demonstrated different peaks of mRNA expressions suggesting that the down-regulation phase of the inflammatory process was controlled by several factors working at different phases.
Insights
This study tracked cytokine mRNA expression in mouse eyes infected with herpes simplex virus type 1 (HSV-1). Key inflammatory markers mirrored disease activity, with immunosuppressive cytokines showing distinct expression patterns.
Area of Science:
- Ophthalmology
- Immunology
- Virology
Background:
- Herpes simplex virus type 1 (HSV-1) can cause ocular inflammation.
- Understanding the immune response in HSV-1 retinitis is crucial for treatment.
Purpose of the Study:
- To investigate cytokine mRNA expression in the contralateral eye during experimental HSV-1 induced ocular inflammation.
- To correlate cytokine expression kinetics with the inflammatory process in HSV-1 retinitis.
Main Methods:
- BALB/c mice were inoculated with HSV-1 in the anterior chamber.
- mRNA from posterior eye segments of uninoculated eyes was analyzed at various time points post-inoculation.
- Reverse transcription-polymerase chain reaction (RT-PCR) was used to quantify mRNA expression of various cytokines and iNOS.
Main Results:
- Induced nitric oxide synthase (iNOS) mRNA peaked at 14 days post-inoculation.
- Pro-inflammatory cytokines (IL-1beta, IL-2, IL-4, IL-10, IFN-gamma, TNFalpha) showed similar expression patterns to iNOS.
- Immunosuppressive cytokines (TGFbeta2, IL-12p35, IL-12p40) exhibited a reverse expression pattern.
Conclusions:
- Cytokine mRNA expression kinetics correlated with the clinicopathological activity of experimental murine HSV-1 retinitis.
- Distinct expression peaks for TGFbeta2 and IL-10 suggest multiple factors regulate the inflammatory down-regulation phase.
- This study provides insights into the temporal immune response during HSV-1 induced ocular inflammation.
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