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Original Article

Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components

Jhang Ho Pak1,*, Ji-Yun Lee2, Bo Young Jeon1, Fuhong Dai2,3, Won Gi Yoo2, Sung-Jong Hong2,*
The Korean Journal of Parasitology 2019;57(4):379-387.
Published online: August 31, 2019

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*Corresponding authors (jhpak@amc.seoul.kr; hongsj@cau.ac.kr)
• Received: July 4, 2019   • Revised: July 27, 2019   • Accepted: August 1, 2019

Copyright © 2019 by The Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components
Korean J Parasitol. 2019;57(4):379-387.   Published online August 31, 2019
DOI: https://doi.org/10.3347/kjp.2019.57.4.379
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Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components
Korean J Parasitol. 2019;57(4):379-387.   Published online August 31, 2019
DOI: https://doi.org/10.3347/kjp.2019.57.4.379
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Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components
Image Image Image
Fig. 1 Bacterial expression and purification of recombinant Cs proteins. (A) Recombinant CsTrip1 purified using a Ni-NTA column in 10% SDS-PAGE. 2.5 μl of elute (lane 1), 1.25 μl of elute (lane 2). (B) Recombinant CsLeg in 10% SDS-PAGE. Uninduced total (lane 1), induced total (lane 2), induced soluble (lane 3), induced pellet (Lane 4), pass-through (lane 5), washing (lane 6), and elute fractions (lane 7 and 8). (C) On-bead cleavage of recombinant CsGrb2 in 10% SDS-PAGE. Uninduced total (lane 1), induced total (lane 2), induced soluble (lane 3), dialyzed soluble (lane 4), pass-through (lane 5), washing (lane 6), cleaved-off CsGrb2 (lane 7), and washed-off Sj26GST tag protein (lane 8).
Fig. 2 Proinflammatory cytokine profiling of HuCCT1 cells in response to C. sinensis ESPs. After 0–24 hr ESP treatment (800 ng/ml), culture supernatants were harvested and analyzed for the production of IL-1β (A), IL-2 (B), IL-6 (C), IL-10 (D), TGF-β1 (E), TGF-β2 (F), TNF-α (G), and INF-γ (H) by ELISA. Data are presented as mean±S.D. of 3 independent experiments. *P<0.05 compared with 0 hr.
Fig. 3 Differential expressions of IL-1β, IL-6, TGF-β1, TGF-β2, and TNF-α in recombinant C. sinensis protein-treated HuCCT1 cells. Cells were treated with 2 μg/ml of CsTrip1, CsLeg, or CsGrb2 proteins for 0–24 hr, and the production of IL-1β (A), IL-6 (B), TGF-β1 (C), TGF-β2 (D), and TNF-α (F) in culture supernatants was determined by ELISA. Data are presented as mean±S.D. of 3 independent experiments. *P<0.05 compared with 0 hr.

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Cytokine Production in Cholangiocarcinoma Cells in Response to Clonorchis sinensis Excretory-Secretory Products and Their Putative Protein Components
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