Characterization of a Shiga-Toxin 1-Resistant Stock of Vero Cells

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 tha...

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Bibliographic Details
Published in:Microbiology and immunology 2004-01, Vol.48 (5), p.377-387
Main Authors: Sekino, Takaomi, Kiyokawa, Nobutaka, Taguchi, Tomoko, Takenouchi, Hisami, Matsui, Jun, Tang, Wei-Ran, Suzuki, Toyo, Nakajima, Hideki, Saito, Masahiro, Ohmi, Kazuhiro, Katagiri, Yohko U., Okita, Hajime, Nakao, Hiroshi, Takeda, Tae, Fujimoto, Junichiro
Format: Article
Language:English
Subjects:
Animals
Butyrates - pharmacology
Cell Survival
Cercopithecus aethiops
Cytotoxins - metabolism
Cytotoxins - toxicity
Globosides - analysis
globotriaocyl ceramide
Microscopy, Confocal
Protein Transport
shiga toxin
Shiga Toxin 1 - metabolism
Shiga Toxin 1 - toxicity
toxin-resistant
Trihexosylceramides - analysis
Trihexosylceramides - immunology
Vero Cells
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Summary: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‐Vero 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.
ISSN:0385-5600
1348-0421
DOI:10.1111/j.1348-0421.2004.tb03527.x