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Published on: March 15, 2012
A novel silkworm infection model with fluorescence imaging using transgenic Trichosporon asahii expressing eGFP
Yasuhiko Matsumoto1, Hideki Yamazaki2, Yusuke Yamasaki2
1Department of Microbiology, Meiji Pharmaceutical University, 2-522-1, Noshio, Kiyose, Tokyo, 204-8588, Japan. ymatsumoto@my-pharm.ac.jp.
Abstract:
Trichosporon asahii is a pathogenic fungus that causes deep mycosis in patients with neutropenia. Establishing an experimental animal model for quantitatively evaluating pathogenicity and developing a genetic recombination technology will help to elucidate the infection mechanism of T. asahii and promote the development of antifungal drugs. Here we established a silkworm infection model with a transgenic T. asahii strain expressing eGFP. Injecting T. asahii into silkworms eventually killed the silkworms. Moreover, the administration of antifungal agents, such as amphotericin B, fluconazole, and voriconazole, prolonged the survival time of silkworms infected with T. asahii. A transgenic T. asahii strain expressing eGFP was obtained using a gene recombination method with Agrobacterium tumefaciens. The T. asahii strain expressing eGFP showed hyphal formation in the silkworm hemolymph. Both hyphal growth and the inhibition of hyphal growth by the administration of antifungal agents were quantitatively estimated by monitoring fluorescence. Our findings suggest that a silkworm infection model using T. asahii expressing eGFP is useful for evaluating both the pathogenicity of T. asahii and the efficacy of antifungal drugs.
Insights
A novel silkworm model using Trichosporon asahii expressing eGFP effectively evaluates fungal pathogenicity and antifungal drug efficacy. This research aids in understanding deep mycosis and developing new treatments for neutropenic patients.
Area of Science:
- Medical Mycology
- Infectious Diseases
- Animal Models
Background:
- Trichosporon asahii causes deep mycosis in immunocompromised patients.
- Developing effective animal models is crucial for studying T. asahii pathogenicity and antifungal drug development.
Purpose of the Study:
- To establish a silkworm infection model for quantitative evaluation of T. asahii pathogenicity.
- To develop a transgenic T. asahii strain for monitoring infection and drug efficacy.
- To assess the utility of the silkworm model in antifungal drug screening.
Main Methods:
- Generation of a transgenic T. asahii strain expressing enhanced green fluorescent protein (eGFP) via Agrobacterium tumefaciens-mediated gene recombination.
- Infection of silkworms with T. asahii and administration of antifungal agents (amphotericin B, fluconazole, voriconazole).
- Quantitative assessment of fungal hyphal growth and inhibition by monitoring eGFP fluorescence in silkworm hemolymph.
Main Results:
- Silkworm injection with T. asahii led to mortality, confirming pathogenicity.
- Antifungal drug administration significantly prolonged silkworm survival.
- eGFP fluorescence allowed real-time, quantitative monitoring of T. asahii hyphal growth and its inhibition by antifungal agents.
Conclusions:
- The silkworm model using eGFP-expressing T. asahii is a valuable tool for assessing fungal pathogenicity.
- This model enables quantitative evaluation of antifungal drug efficacy.
- The established model can accelerate the development of novel antifungal therapies for T. asahii infections.
