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The gene expression and proteomic profiling of Acanthamoeba isolates
Chayan Sharma1, Sumeeta Khurana1, Alka Bhatia2
1Department of Medical Parasitology, Postgraduate Institute of Medical Education & Research, Chandigarh, 160012, India.
Experimental Parasitology
|October 11, 2023
Summary
This study identifies key genes like phospholipase and lysophospholipase involved in Acanthamoeba pathogenesis, crucial for understanding Acanthamoeba keratitis and encephalitis. Proteomic analysis further reveals differential protein expression in pathogenic versus non-pathogenic isolates.
Area of Science:
- Microbiology
- Molecular Biology
- Parasitology
Background:
- Acanthamoeba is a protozoan parasite responsible for severe eye infections (Acanthamoeba keratitis) and brain infections (granulomatous amoebic encephalitis).
- Understanding Acanthamoeba pathogenesis requires investigating its interactions with the host immune system.
- Genomic and proteomic analyses are essential for identifying virulence factors and pathogenic mechanisms.
Purpose of the Study:
- To identify potential pathogenic genes and protein targets in Acanthamoeba.
- To analyze the differential gene expression in pathogenic versus non-pathogenic Acanthamoeba isolates.
- To investigate the impact of PHMB treatment on protein expression in Acanthamoeba.
Main Methods:
- Selection and validation of candidate pathogenic genes (lysophospholipase, phospholipase, S8/S53 peptidase, carboxylesterase, mannose-binding protein) using conventional and real-time PCR.
- Evaluation of relative gene expression in animal models of Acanthamoeba keratitis and encephalitis.
- Proteomic analysis using LC-MS/MS on pathogenic and non-pathogenic isolates before and after PHMB treatment.
Main Results:
- Conventional PCR confirmed the presence of selected genes in clinical and environmental Acanthamoeba isolates.
- Significant upregulation of phospholipase, lysophospholipase, and mannose-binding genes was observed in the keratitis isolate during Acanthamoeba keratitis.
- Phospholipase, lysophospholipase, S8/S53 peptidase, and carboxylesterase genes were upregulated in the encephalitis isolate.
- Proteomic data showed differential protein expression between pathogenic and non-pathogenic isolates, with and without PHMB treatment.
Conclusions:
- Lysophospholipase, phospholipase, S8/S53 peptidase, carboxylesterase, and mannose-binding protein are implicated in Acanthamoeba pathogenesis.
- Proteomic profiling highlights proteins critical for parasite growth, survival, and virulence.
- The findings provide a foundation for future studies, including knockout experiments, to further elucidate Acanthamoeba virulence mechanisms.
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