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. 2019 Jun;57(3):217-223.
doi: 10.3347/kjp.2019.57.3.217. Epub 2019 Jun 30.

Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts

Hae-Jin Sohn  1 , Ga-Eun Seo  1 , Jae-Ho Lee  1 , A-Jeong Ham  1 , Young-Hwan Oh  1 , Heekyoung Kang  1 , Ho-Joon Shin  1
Affiliations

Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts

Hae-Jin Sohn et al. Korean J Parasitol. 2019 Jun.

Abstract

Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of IL-1α, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of IL-1α, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

Keywords: Acanthamoeba castellanii; acanthamoebic keratitis; cytotoxicity; human corneal epithelial cell; pro-inflammatory cytokine.

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Conflict of interest statement

CONFLICT OF INTEREST

The authors declare no conflict of interest related to this study.

Figures

Fig. 1
Fig. 1
Morphologic change of A. castellanii trophozoites into pre- and mature cysts by induction with encystment medium. Arrows indicate double-walled mature cysts. Scale bar=20 μm.
Fig. 2
Fig. 2
Cytopathic changes of HCECs co-cultured with A. castellanii. T, trophozoite only; T/C, trophozoite and cyst; C, cyst only. Scale bar=20 μm.
Fig. 3
Fig. 3
In vitro cytotoxicity of A. castellanii to HCECs by LDH assay. Co-cultured with trophozoites and cysts. *P<0.005.
Fig. 4
Fig. 4
Expression of cytokines in HCECs co-cultured with A. castellanii trophozoites or cysts. *P<0.005.
Fig. 5
Fig. 5
Secretion of IL-6 (A) and IL-8 (B) from HCECs co-cultured with A. castellanii trophozoites or cysts. *P<0.005.
See this image and copyright information in PMC

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