Characterization of a Shiga-toxin 1-resistant stock of Vero cells

Takaomi Sekino, Nobutaka Kiyokawa, Tomoko Taguchi, Hisami Takenouchi, Jun Matsui, Wei Ran Tang, Toyo Suzuki, Hideki Nakajima, Masahiro Saito, Kazuhiro Ohmi, Yohko U. Katagiri, Hajime Okita, Hiroshi Nakao, Tae Takeda, Junichiro Fujimoto

Research output: Contribution to journal › Article

15 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)377-387
Number of pages11
JournalMicrobiology and Immunology
Volume48
Issue number5
Publication statusPublished - 2004
Externally publishedYes

Fingerprint

Shiga Toxin 1
Vero Cells
Shiga Toxins
Lipids
Glycosphingolipids
Butyrates
Bromides
Confocal Microscopy

Keywords

  • Globotriaocyl ceramide
  • Shiga toxin
  • Toxin-resistant
  • Vero cells

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Microbiology (medical)
  • Microbiology

Cite this

Sekino, T., Kiyokawa, N., Taguchi, T., Takenouchi, H., Matsui, J., Tang, W. R., ... Fujimoto, J. (2004). Characterization of a Shiga-toxin 1-resistant stock of Vero cells. Microbiology and Immunology, 48(5), 377-387.

Characterization of a Shiga-toxin 1-resistant stock of Vero cells. / 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.

In: Microbiology and Immunology, Vol. 48, No. 5, 2004, p. 377-387.

Research output: Contribution to journal › Article

Sekino, T, Kiyokawa, N, Taguchi, T, Takenouchi, H, Matsui, J, Tang, WR, Suzuki, T, Nakajima, H, Saito, M, Ohmi, K, Katagiri, YU, Okita, H, Nakao, H, Takeda, T & Fujimoto, J 2004, 'Characterization of a Shiga-toxin 1-resistant stock of Vero cells', Microbiology and Immunology, vol. 48, no. 5, pp. 377-387.
Sekino T, Kiyokawa N, Taguchi T, Takenouchi H, Matsui J, Tang WR et al. Characterization of a Shiga-toxin 1-resistant stock of Vero cells. Microbiology and Immunology. 2004;48(5):377-387.
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. / Characterization of a Shiga-toxin 1-resistant stock of Vero cells. In: Microbiology and Immunology. 2004 ; Vol. 48, No. 5. pp. 377-387.
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abstract = "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.",
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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

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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.

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EP - 387

JO - Microbiology and Immunology

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