Real-Time Cell Analysis for Monitoring Cholera Toxin-Induced Human Intestinal Epithelial Cell Response

详细信息    查看全文

• 作者：Julian Ye ; Yun Luo ; Weijia Fang ; Junhang Pan ; Zheng Zhang…
• 刊名：Current Microbiology
• 出版年：2015
• 出版时间：April 2015
• 年：2015
• 卷：70
• 期：4
• 页码：536-543
• 全文大小：634 KB
• 参考文献：1. Atienza, JM, Zhu, J, Wang, X, Xu, X, Abassi, Y (2005) Dynamic monitoring of cell adhesion and spreading on microelectronic sensor arrays. J Biomol Screen 10: pp. 795-805 CrossRef
  2. Bernas, T, Dobrucki, J (2002) Mitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1, and NAO mitochondrial fluorescent probes. Cytometry 47: pp. 236-242 CrossRef
  3. Chinnapen, DJ, Chinnapen, H, Saslowsky, D, Lencer, WI (2007) Rafting with cholera toxin: endocytosis and trafficking from plasma membrane to ER. FEMS Microbiol Lett 266: pp. 129-137 CrossRef
  4. Haan, L, Hirst, TR (2004) Cholera toxin: a paradigm for multi-functional engagement of cellular mechanisms (Review). Mol Membr Biol 21: pp. 77-92 CrossRef
  5. Donta, ST (1974) Differentiation between the steroidogenic effects of cholera enterotoxin and adrenocorticotropin through use of a mutant adrenal cell line. J Infect Dis 129: pp. 728-731 CrossRef
  6. Guerrant, RL, Brunton, LL, Schnaitman, TC, Rebhun, LI, Gilman, AG (1974) Cyclic adenosine monophosphate and alteration of Chinese hamster ovary cell morphology: a rapid, sensitive in vitro assay for the enterotoxins of Vibrio cholerae and Escherichia coli. Infect Immun 10: pp. 320-327
  7. He, X, Wang, J, Steele, J, Sun, X, Nie, W, Tzipori, S, Feng, H (2009) An ultrasensitive rapid immunocytotoxicity assay for detecting Clostridium difficile toxins. J Microbiol Methods 78: pp. 97-100 CrossRef
  8. Holmes, RK, Twiddy, EM (1983) Characterization of monoclonal antibodies that react with unique and cross-reacting determinants of cholera enterotoxin and its subunits. Infect Immun 42: pp. 914-923
  9. Holmgren, J (1981) Actions of cholera toxin and the prevention and treatment of cholera. Nature 292: pp. 413-417 CrossRef
  10. Irelan, JT, Wu, MJ, Morgan, J, Ke, N, Xi, B, Wang, X, Xu, X, Abassi, YA (2011) Rapid and quantitative assessment of cell quality, identity, and functionality for cell-based assays using real-time cellular analysis. J Biomol Screen 16: pp. 313-322 CrossRef
  11. Jin, D, Luo, Y, Zheng, M, Li, H, Zhang, J, Stampfl, M, Xu, X, Ding, G, Zhang, Y, Tang, YW (2013) Quantitative detection of Vibrio cholera toxin by real-time and dynamic cytotoxicity monitoring. J Clin Microbiol 51: pp. 3968-3974 CrossRef
  12. Kanwar, JR, Kanwar, RK (2009) Gut health immunomodulatory and anti-inflammatory functions of gut enzyme digested high protein micro-nutrient dietary supplement-Enprocal. BMC Immunol 10: pp. 7 CrossRef
  13. Kaper, JB, Morris, JG, Levine, MM (1995) Cholera. Clin Microbiol Rev 8: pp. 48-86
  14. Krysko, DV, Vanden Berghe, T, D’Herde, K, Vandenabeele, P (2008) Apoptosis and necrosis: detection, discrimination and phagocytosis. Methods 44: pp. 205-221 CrossRef
  15. Krysko, DV, Vanden Berghe, T, Parthoens, E, D’Herde, K, Vandenabeele, P (2008) Methods for distinguishing apoptotic from necrotic cells and measuring their clearance. Methods Enzymol 442: pp. 307-341 CrossRef
  16. Lockshin, RA, Zakeri, Z (2004) Apoptosis, autophagy, and more. Int J Biochem Cell Biol 36: pp. 2405-2419 CrossRef
  17. Martinez-Serra, J, Gutierrez, A, Munoz-Capo, S, Navarro-Palou, M, Ros, T, Amat, JC, Lopez, B, Marcus, TF, Fueyo, L, Suquia, AG, Gines, J, Rubio, F, Ramos, R, Besalduch, J (2014) xCELLigence system for real-time label-free monitoring of growth and viability of cell lines from hematologi
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  
• 出版者：Springer New York
• ISSN：1432-0991

文摘

The pathogenic mechanism of Vibrio cholerae manifests as diarrhea and causes life-threatening dehydration. Here, we observe the human intestinal epithelial cells (HIEC) response to Cholera toxin (CT) by a real-time cell analysis (RTCA) platform, and disclose the difference from CT-induced cytotoxicity and others in HIEC. An HIEC cell of 1.0?×?105?cells/mL was characterized as the suitable concentration for each well. For experimentation, the assay requires an inoculation of CT dissolved in Dulbecco’s phosphate-buffered saline with 0.1?% gelatin for a period of 18-5?h. The dimensionless impedance cell index curve presented characteristic dose- and time-dependent drop responses at the first stage, and the CT-induced cytotoxicity was the most remarkable following exposure for 18-5?h (P?=?0.0002). Following the obvious cytotoxic reaction, the CI curve gradually increased over time until the original CI value, indicating that self-recovery occurred. The CT-induced CI curve for HIEC was different from that induced by other toxins, including diphtheria and Clostridium difficile toxin. Collectively, these results suggest that the CT-induced cytotoxicity in HIEC was absolutely different from that induced by C. difficile and other toxins because of the different pathogeneses that were correlated with the specific CI curve generated by the RTCA system. In summary, our data show that the assay described here is a convenient and rapid high-throughput tool for real-time monitoring of host cellular responses to CT on the basis of the characteristic CI curve.