Korean J Parasitol.  2011 Mar;49(1):1-8. 10.3347/kjp.2011.49.1.1.

Phospholipase Activities in Clinical and Environmental Isolates of Acanthamoeba

• Matin A
• Jung SY

• KMID: 1715626
• DOI: http://doi.org/10.3347/kjp.2011.49.1.1

Abstract

The pathogenesis and pathophysiology of Acanthamoeba infections remain incompletely understood. Phos-pholipases are known to cleave phospholipids, suggesting their possible involvement in the host cell plasma membrane disruption leading to host cell penetration and lysis. The aims of the present study were to determine phospholipase activities in Acanthamoeba and to determine their roles in the pathogenesis of Acanthamoeba. Using an encephalitis isolate (T1 genotype), a keratitis isolate (T4 genotype), and an environmental isolate (T7 genotype), we demonstrated that Acanthamoeba exhibited phospholipase A2 (PLA2) and phospholipase D (PLD) activities in a spectrophotometry-based assay. Interestingly, the encephalitis isolates of Acanthamoeba exhibited higher phospholipase activities as compared with the keratitis isolates, but the environmental isolates exhibited the highest phospholipase activities. Moreover, Acanthamoeba isolates exhibited higher PLD activities compared with the PLA2. Acanthamoeba exhibited optimal phospholipase activities at 37degrees C and at neutral pH indicating their physiological relevance. The functional role of phospholipases was determined by in vitro assays using human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. We observed that a PLD-specific inhibitor, i.e., compound 48/80, partially inhibited Acanthamoeba encephalitis isolate cytotoxicity of the host cells, while PLA2-specific inhibitor, i.e., cytidine 5'-diphosphocholine, had no effect on parasite-mediated HBMEC cytotoxicity. Overall, the T7 exhibited higher phospholipase activities as compared to the T4. In contract, the T7 exhibited minimal binding to, or cytotoxicity of, HBMEC.

Keyword

Acanthamoeba; human brain microvascular endothelial cell; phospholipase A2; phospholipase D; adhesion; cytotoxicity

MeSH Terms

Acanthamoeba/*enzymology/genetics/*isolation & purification/physiology
Cell Adhesion
Cells, Cultured
Endothelial Cells/parasitology
Humans
Keratitis/*parasitology
Phospholipase D/genetics/*metabolism
Phospholipases A2/genetics/*metabolism
Protozoan Proteins/genetics/*metabolism
Soil/*parasitology