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 language | English |
|---|---|
| Pages (from-to) | 377-387 |
| Number of pages | 11 |
| Journal | Microbiology and Immunology |
| Volume | 48 |
| Issue number | 5 |
| Publication status | Published - 2004 |
| Externally published | Yes |
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Keywords
- Globotriaocyl ceramide
- Shiga toxin
- Toxin-resistant
- Vero cells
ASJC Scopus subject areas
- Immunology and Microbiology(all)
- Microbiology (medical)
- Microbiology
Cite this
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
}
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
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
UR - http://www.scopus.com/inward/record.url?scp=2442655048&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=2442655048&partnerID=8YFLogxK
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 -