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Updated: Mar 24, 2026
Recurrent Herpetic Stromal Keratitis in Mice, a Model for Studying Human HSK
Published on: December 18, 2012
Experimental modeling of Acanthamoeba keratitis in mice: Isolation and cultivation of Acanthamoeba castellanii
Hae-Jin Sohn1, Suk-Yul Jung2, Jong-Hyun Kim3
1Department of Microbiology, Ajou University School of Medicine, And Department of Biomedical Science, Graduate School of Ajou University, Suwon, 16499, Republic of Korea.
Abstract:
Free-living Acanthamoeba can lead to granulomatous amoebic encephalitis or Acanthamoeba keratitis (AK), an eye infection. AK is most common among contact lens wearers, and causes include long-term lens use, contamination of the contact lens, and corneal trauma. In vitro studies of AK, especially the development of therapeutic drugs, need to be confirmed by in vivo experiments. We developed our AK mouse model using a variety of methods. A. castellanii (AK/AJ1; 1 × 106 cells) were loaded onto 2 mm contact lens pieces for insertion into the scratched eyes of mice under anesthesia, and the eyelids were sutured. After infection (1 to 14 days), in the daily follow-up, it was observed that the AK lesion had progressed in the mouse eyes, and PCR confirmed the amplification of the Acanthamoeba DNA. This study investigated the effective cultivation method to certify PCR analysis in the AK mouse model. After inoculation at 1, 3, 7, and 14 days, PCR was performed on eyeball samples collected from the AK mouse model. Successful cultivation from homogenized eyeballs of A. castellanii was done in the non-nutrient agar with the lawn of Escherichia coli and scaled up with PYG medium. Furthermore, the PCR products obtained from the amoeba cultured in the AK model showed a match of over 99 % with the genetic information of the inoculated amoeba. We suggest that cultivation and PCR methods can confirm the development of AK in the AK mouse model.
Insights
Researchers developed a mouse model for Acanthamoeba keratitis (AK), an eye infection. Cultivation and PCR methods effectively confirmed AK development in the model, aiding future therapeutic research.
Area of Science:
- Ophthalmology
- Infectious Diseases
- Microbiology
Background:
- Free-living Acanthamoeba can cause severe eye infections like Acanthamoeba keratitis (AK).
- AK is prevalent in contact lens wearers due to factors like prolonged use, contamination, and corneal trauma.
- In vitro studies for AK therapeutics require validation through in vivo models.
Purpose of the Study:
- To establish and validate an Acanthamoeba keratitis (AK) mouse model.
- To investigate effective cultivation and PCR methods for confirming AK development in vivo.
Main Methods:
- Developed an AK mouse model by inserting Acanthamoeba castellanii-loaded contact lenses into scratched mouse eyes.
- Monitored lesion progression daily and performed PCR on eyeball samples at 1, 3, 7, and 14 days post-infection.
- Cultivated Acanthamoeba from homogenized eyeballs using non-nutrient agar with Escherichia coli, scaled up with PYG medium.
Main Results:
- The AK mouse model demonstrated progressive lesion development over 14 days.
- PCR successfully amplified Acanthamoeba DNA from infected mouse eyes.
- Cultivation methods yielded viable Acanthamoeba from eyeball samples.
- PCR analysis of cultured amoebas showed >99% genetic match with the inoculated strain.
Conclusions:
- The developed AK mouse model effectively replicates the infection.
- Combined cultivation and PCR methods reliably confirm AK development in the mouse model.
- This validated model supports further research into AK pathogenesis and treatment.
