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Quantitative proteomic analysis and functional characterization of Acanthamoeba castellanii exosome-like vesicles

Wei-Chen Lin1,2, Chia-Yun Tsai1, Jian-Ming Huang3

  • 1Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan City, 701, Taiwan.

Parasites & Vectors
|October 11, 2019
PubMed
Abstract

Insights

Extracellular vesicles (EVs) from Acanthamoeba castellanii mediate host cell damage and immune responses. Proteomic analysis reveals hydrolases and oxidoreductases within these parasite EVs, offering insights into disease mechanisms.

Area of Science:

  • Parasitology
  • Cell Biology
  • Proteomics

Background:

  • Pathogenic protozoans, like Acanthamoeba castellanii, utilize extracellular vesicles (EVs) for communication and host manipulation.
  • A. castellanii causes severe keratitis and fatal granulomatous encephalitis.
  • The specific roles and contents of parasite-derived EVs in Acanthamoeba pathogenesis remain largely unknown.

Purpose of the Study:

  • To characterize the protein composition of Acanthamoeba castellanii EVs.
  • To investigate the functional roles of these EVs in host cell interactions, including cytotoxicity and immune response.
  • To elucidate potential mechanisms of Acanthamoeba pathogenesis mediated by EVs.

Main Methods:

  • Purification and confirmation of A. castellanii EVs using electron microscopy and nanoparticle tracking analysis.
  • Assessment of EV-induced cytotoxicity and immune responses in host cells (rat glioma C6 and human THP-1 cells).
  • Quantitative proteomic analysis of EV protein content via LC-MS/MS.

Main Results:

  • A. castellanii EVs were observed to fuse with host C6 cells.
  • Parasite EVs triggered immune responses in THP-1 cells and exhibited cytotoxicity towards C6 cells.
  • Proteomic analysis identified 130 proteins, with hydrolases (50.2%) and oxidoreductases (31.7%) being the most abundant families. Aminopeptidase activity was confirmed in the EVs.

Conclusions:

  • Proteomic profiling and functional characterization of A. castellanii EVs offer deep insights into their molecular cargo.
  • These findings highlight the potential mechanisms by which EVs contribute to the pathogenesis of Acanthamoeba infections.
  • Understanding EV components and functions is crucial for developing strategies against Acanthamoeba-related diseases.
Keywords:
AcanthamoebaExosomeExtracellular vesicles

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