TY - JOUR
T1 - Characterization of a Shiga-toxin 1-resistant stock of Vero cells
AU - Sekino, Takaomi
AU - Kiyokawa, Nobutaka
AU - Taguchi, Tomoko
AU - Takenouchi, Hisami
AU - Matsui, Jun
AU - Tang, Wei Ran
AU - Suzuki, Toyo
AU - Nakajima, Hideki
AU - Saito, Masahiro
AU - Ohmi, Kazuhiro
AU - Katagiri, Yohko U.
AU - Okita, Hajime
AU - Nakao, Hiroshi
AU - Takeda, Tae
AU - Fujimoto, Junichiro
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2004
Y1 - 2004
N2 - Shiga toxins (Stxs, also referred to as verotoxins) were first described as a novel cytotoxic activity against Vero cells. In this study, we report the characterization of an Stx1-resistant (R-) stock of Vero cells. (1) When the susceptibility of R-Vero cells to Stx1 cytotoxicity was compared to that of Stx1-sensitive (S-) Vero cells by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, cell viability after 48-hr exposure to 10 pg/ml of Stx1 was greater than 80% and less than 15%, respectively. (2) Although both a binding assay of fluorescence-labeled Stx1 and lipid analysis indicated considerable expression of Gb3Cer, a functional receptor for Stxs, in both Vero cells, anti-Gb3Cer monoclonal antibodies capable of binding to S-Vero cells failed to effectively label R-Vero cells, suggesting a conformational difference in the Gb3Cer expressed on R-Vero cells. (3) The lipid analysis also showed that the R-Vero cells contained significant amounts of Gb4Cer. In addition, introduction of exogenous Gb4Cer into S-Vero cells slightly inhibited Stx1 cytotoxicity, suggesting some-correlation between glycosphingolipid composition and Stx1 resistance. (4) Both butyrate treatment and serum depression eliminated the Stx1 resistance of R-Veto cells. (5) The results of the analysis by confocal microscopy suggest a difference in intracellular transport of Stx1 between R-Vero and S-Vero cells. Further study of R-Vero cells may provide a model of Stx1 resistance via distinct intracellular transport of Stx1.
AB - Shiga toxins (Stxs, also referred to as verotoxins) were first described as a novel cytotoxic activity against Vero cells. In this study, we report the characterization of an Stx1-resistant (R-) stock of Vero cells. (1) When the susceptibility of R-Vero cells to Stx1 cytotoxicity was compared to that of Stx1-sensitive (S-) Vero cells by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, cell viability after 48-hr exposure to 10 pg/ml of Stx1 was greater than 80% and less than 15%, respectively. (2) Although both a binding assay of fluorescence-labeled Stx1 and lipid analysis indicated considerable expression of Gb3Cer, a functional receptor for Stxs, in both Vero cells, anti-Gb3Cer monoclonal antibodies capable of binding to S-Vero cells failed to effectively label R-Vero cells, suggesting a conformational difference in the Gb3Cer expressed on R-Vero cells. (3) The lipid analysis also showed that the R-Vero cells contained significant amounts of Gb4Cer. In addition, introduction of exogenous Gb4Cer into S-Vero cells slightly inhibited Stx1 cytotoxicity, suggesting some-correlation between glycosphingolipid composition and Stx1 resistance. (4) Both butyrate treatment and serum depression eliminated the Stx1 resistance of R-Veto cells. (5) The results of the analysis by confocal microscopy suggest a difference in intracellular transport of Stx1 between R-Vero and S-Vero cells. Further study of R-Vero cells may provide a model of Stx1 resistance via distinct intracellular transport of Stx1.
KW - Globotriaocyl ceramide
KW - Shiga toxin
KW - Toxin-resistant
KW - Vero cells
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U2 - 10.1111/j.1348-0421.2004.tb03527.x
DO - 10.1111/j.1348-0421.2004.tb03527.x
M3 - Article
C2 - 15215625
AN - SCOPUS:2442655048
VL - 48
SP - 377
EP - 387
JO - Microbiology and Immunology
JF - Microbiology and Immunology
SN - 0385-5600
IS - 5
ER -