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The pathogenesis and pathophysiology of Acanthamoeba infections remain incompletely understood. Phospholipases 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 37˚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.
목차
Abstract
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
참고문헌 (28)
참고문헌 신청
1. [학술지(정기간행물)] - Khan NA / 2006 / Acanthamoeba: biology and increasing importance in human health / FEMS Microbiol Rev 30 (564) : 564 ~ 595
2. [학술지(정기간행물)] - Marciano-Cabral F / 2003 / Acanthamoeba spp. as agents of disease in humans / Clin Microbiol Rev 16 : 273 ~ 307
3. [학술지(정기간행물)] - Schuster FL / 2004 / Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals / Int J Parasitol 34 : 1001 ~ 1027
4. [학술지(정기간행물)] - Jung SY / 2008 / Pathogen–pathogen interactions: a comparative study of Escherichia coli interactions with the clinical and environmental isolates of Acanthamoeba / World J Microbiol Biotechnol 24 : 2339 ~ 2348
5. [학술지(정기간행물)] - Victoria EJ / 1975 / Plasma membrane and soluble lysophospholipases of Acanthamoeba castellanii / Arch Biochem Biophys 171 : 255 ~ 258
2. [학술지(정기간행물)] - Marciano-Cabral F / 2003 / Acanthamoeba spp. as agents of disease in humans / Clin Microbiol Rev 16 : 273 ~ 307
3. [학술지(정기간행물)] - Schuster FL / 2004 / Free-living amoebae as opportunistic and non-opportunistic pathogens of humans and animals / Int J Parasitol 34 : 1001 ~ 1027
4. [학술지(정기간행물)] - Jung SY / 2008 / Pathogen–pathogen interactions: a comparative study of Escherichia coli interactions with the clinical and environmental isolates of Acanthamoeba / World J Microbiol Biotechnol 24 : 2339 ~ 2348
5. [학술지(정기간행물)] - Victoria EJ / 1975 / Plasma membrane and soluble lysophospholipases of Acanthamoeba castellanii / Arch Biochem Biophys 171 : 255 ~ 258
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UCI(KEPA) : I410-ECN-0101-2014-510-002511717