Cyclosporine A and FK506 as Potent Inhibitors of Streptococcus intermedius Intermedilysin-Induced NFAT-1 Activation

Heni Susilowati, Hirohiko Okamura, Katsuhiko Hirota, Kaya Yoshida, Atsushi Tabata, Hideaki Nagamune, Tatsuji Haneji, Yoichiro Miyake


Cyclosporine A (CsA) and tacrolimus (FK506), a member of calcineurin inhibitors, inhibit inflammation process as part of immune response. Nuclear activated T cells subfamily NFAT1 is a trascription factor responsible for the regulation of immune response genes. Streptococcus intermedius, an oral commensal bacterium, has been shown to strongly associate with liver abscess.  The S. intermedius strains produce intermedilysin (ILY), which is responsible for the bacterial virulence. Cyclosporine A and FK506 have been widely used to control NFAT activation in most of cell types, however the ability of CsA and FK506 to inhibit ILY-induced NFAT1 activation remains to be investigated. The aim of this study was to investigate the effect of CsA and FK506 on NFAT1 activation caused by ILY. Human cholangiocellular cell line HuCCT1 was stimulated with various concentrations of ILY. The cell and nuclear morphological change was observed by microscopy analysis. The NFAT1 nuclear translocation that indicates its activation was detected by immunocytochemistry. The inhibitory effect of CsA and FK506 was tested after 30 min application before ILY treatment by using immunofluorescence microscope. The results showed cell and nuclear shrinkage in ILY-treated cells. The NFAT1 was translocated to the nuclei in HuCCT1 cells, and observed in dose dependent manner.  Cyclosporine A and FK506 inhibited ILY-induced NFAT1 nuclear translocation.  In conclusion, CsA and FK506 may act as potent inflammation control agents in S. intermedius ILY-infected cells.

Keywords: Cyclosporine A, FK506, NFAT1, intermedilysin

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Addy, N.A., E.F. Fornasiero, T.R. Stevens, J.R. Taylor, M.R. Picciotto. 2007. Role of calcineurin in nicotine-mediated locomotor sensitization. J. Neurosci. 27: 8571-8580.

Almawi, W.Y., O.K. Melemedjian. 2000. Clinical and mechanistic differences between FK506 (tacrolimus) and cyclosporine A. Nephrol. Dial. Transplant. 15 (2000) 1916-1918.

Dolmetsch, R.E., R.S. Lewis, C.C. Goodnow, J.I. Healy. 1997. Differential activation of transcription factors induced by Ca2+ response amplitude and duration. Nature 386: 855-858.

Jain, J., P.G. McCaffrey, Z. Miner, T.K. Kerppola, J.N. Lambert, G.L. Verdine, T. Curran, A. Rao. 1993. The T-cell transcription factor NFATp is a substrate for calcineurin and interacts with Fos and Jun. Nature 365: 352-355.

Liu, J., J.D. Farmer Jr, W.S. Lane, J. Friedman, I. Weissman, S.L. Schreiber. 1991. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell 66: 807-815.

Ludwig, A., W. Goebel. 2000. Dangerous signals from E. coli toxin. Nat. Med. 6 (2000) 741-742.

Luo, C., K.T.Y. Shaw, A. Raghavan, J. Aramburu, F. Garcia-Cozar, B.A. Perrino, P.G. Hogan, A. Rao. 1996. Interaction of calcineurin with a domain of the transcription factor NFAT1 that controls nuclear import. Proc. Natl. Acad. Sci. USA 93: 8907-8912.

Macey, M.G., R.A. Whiley, L. Miller, H. Nagamune. 2001. Effect on polymorphonuclear cell function of a human-specific cytotoxin, intermedilysin, expressed by Streptococcus intermedius. Infect. Immun. 69: 6102-6109.

Macian, F. 2005. NFAT proteins: key regulators of T-cell development and function. Nat. Rev. Immunol. 5: 472-484.

Nagamune, H., R.A. Whiley, T. Goto, Y. Inai, T. Maeda, J.M. Hardie, H. Kourai. 2000. Distribution of the intermedilysin gene among the anginosus group streptococci and correlation between intermedilysin production and deep-seated infection with Streptococcus intermedius. J. Clin. Microbiol. 38: 220-226.

Nagamune, H., C. Ohnishi, A. Katsuura, K. Fushitani, R.A. Whiley, A. Tsuji, Y. Matsuda. 1996. Intermedilysin, a novel cytotoxin specific for human cells, secreted by Streptococcus intermedius UNS46 isolated from a human liver abscess. Infect. Immun. 64: 3093-3100.

Ruff, V.A., K.L. Leach. 1995. Direct demonstration of NFATp dephosphorylation and nuclear localization in activated HT-2 cells using a specific NFATp polyclonal antibody. J. Biol. Chem. 270: 22602-22607.

Shaw, K.T.Y., A.M. Ho, A. Raghavan, J. Kim, J. Jain, J. Park, S. Sharma, A. Rao, P.G. Hogan. 1995. Immunosuppressive drugs prevent a rapid dephosphorylation of transcription factor NFAT1 in stimulated immune cells. Proc. Natl. Acad. Sci. USA 92: 11205-11209.

Söderblom, T., Å. Laestadius, C. Oxhamre, A. Aperia, A. Richter-Dahlfors. 2002. Toxin-induced calcium oscillations: a novel strategy to affect gene regulation in target cells. Int. J. Med. Microbiol. 291: 511-515.

Sukeno, A, H. Nagamune, R.A. Whiley, S.I. Jafar, J. Aduse-Opoku, K. Ohkura, T. Maeda, K. Hirota, Y. Miyake, H. Kourai. 2005. Intermedilysin is essential for the invasion of hepatoma HepG2 cells by Streptococcus intermedius. Microbiol. Immunol. 49: 681-694.

Wagner, K.W., R. Schön, M. Schumacher, R. Schmelzeisen, D. Schulze. 2006. Case report: brain and liver abscesses caused by oral infection with Streptococcus intermedius. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 102: e21-e23.

Wesselborg, S., D.A. Fruman, J.K. Sagoo, B.E. Bierer, S.J. Burakoff. 1996. Identification of a physical interaction between calcineurin and nuclear factor of activated T cells (NFATp). J. Biol. Chem. 271: 1274-1277.

Whiley, R.A., D. Beighton, T.G. Winstanley, H.Y. Fraser, J.M. Hardie. 1992. Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus (the Streptococcus milleri group): association with different body sites and clinical infections. J. Clin. Microbiol. 30: 243-244.

Whiley, R.A., H. Fraser, J.M. Hardie, D. Beighton. 1990. Phenotypic differentiation of Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus strains within the “Streptococcus milleri group”. J. Clin. Microbiol. 28: 1497-1501.

Wu, H., A. Peisley, I.A. Graef, G.R. Crabtree. 2007. NFAT signaling and the invention of vertebrates. Trends Cell Biol. 17: 251-260.

Yamamoto, M., T. Fukushima, S. Ohshiro, Y. Go, H. Tsugu, K. Kono, M. Tomonaga. 1999. Brain abscess caused by Streptococcus intermedius: two case reports. Surg. Neurol. 51: 219-222.

Yokoyama, K., H. Higashi, S. Ishikawa, Y. Fujii, S. Kondo, H. Kato, T. Azuma, A. Wada, T. Hirayama, H. Aburatani, M. Hatakeyama. 2005. Functional antagonism between Helicobacter pilory CagA and vacuolating toxin VacA in control of the NFAT signaling pathway in gastric epithelial cells. Proc. Natl. Acad. Sci. USA 102: 9661-9666.


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