Induction of apoptosis in Vero cells by Aeromonas veronii biovar sobria vacuolating cytotoxic factor
Luciano Moura Martins
Departamento de Microbiologia e Imunologia, Instituto de Biologia
Search for more papers by this authorCleide Ferreira Catani
Departamento de Microbiologia e Imunologia, Instituto de Biologia
Search for more papers by this authorRosabel Márquez Falcón
Departamento de Microbiologia, Instituto Pedro Kouri, Havana, Cuba
Search for more papers by this authorGleize Villela Carbonell
Bacteriologia Clinica, Instituto Adolfo Lutz, Campinas, SP, Brazil
Search for more papers by this authorAdriano A. Azzoni
Centro de Biologia Molecular e Genética, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil
Search for more papers by this authorTomomasa Yano
Departamento de Microbiologia e Imunologia, Instituto de Biologia
Search for more papers by this authorLuciano Moura Martins
Departamento de Microbiologia e Imunologia, Instituto de Biologia
Search for more papers by this authorCleide Ferreira Catani
Departamento de Microbiologia e Imunologia, Instituto de Biologia
Search for more papers by this authorRosabel Márquez Falcón
Departamento de Microbiologia, Instituto Pedro Kouri, Havana, Cuba
Search for more papers by this authorGleize Villela Carbonell
Bacteriologia Clinica, Instituto Adolfo Lutz, Campinas, SP, Brazil
Search for more papers by this authorAdriano A. Azzoni
Centro de Biologia Molecular e Genética, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil
Search for more papers by this authorTomomasa Yano
Departamento de Microbiologia e Imunologia, Instituto de Biologia
Search for more papers by this authorEditor: Alex van Belkum
Abstract
Recently, a cytotoxin named vacuolating cytotoxic factor (VCF) in Aeromonas sobria and Aeromonas veronii biovar sobria was described. We have now purified this factor using ion metallic affinity chromatography. The VCF is a nonhemolytic enterotoxin that acts as a serine protease. The toxin can be partially neutralized by serum antiaerolysin and it induced not only cytoplasmatic vacuole formation, but also mitochondrial disorders that must be signaling the apoptotic pathways, leading to Vero (African green monkey kidney) cell death. These results suggest that the VCF is a virulence factor of these bacteria, participating in the processes of human disease provoked by preformed toxins in food and infection.
References
- Abrami L, Fivaz M, Decroly E, Seidah MG, Jean TG, Lepper SH, Bucley T & Van Der Goot FG (1998a) The pore-forming toxin pro-aerolysin is activated by furin. J Biol Chem 273: 32656–32661.
- Abrami L, Fivaz M, Glauser PE, Parton RG & Van Der Goot FG (1998b) A pore-forming toxin interacts with GP1-anchored protein and causes vacuolation of the endoplasmatic reticulum. J Cell Biol 140: 525–540.
- Asao T, Kosaki S, Kato Y, Otsu K & Sakakuchi G (1986) Purification and characterization of an Aeromonas hydrophila hemolysin. J Clin Microbiol 24: 228–232.
-
Borenfreund E &
Puerner JA (1984) A simple quantitative procedure using monolayer cultures for cytotoxicity assays.
J Tiss Cult Methods
9: 7–9.
10.1007/BF01666038 Google Scholar
- Cover TL (1996) The vacuolating cytotoxin of Helicobacter pylori. Mol Microbiol 20: 241–246.
- Esteve C & Birkbeck TH (2004) Secretion of haemolysins and proteases by Aeromonas hydrophila EO63: separation and characterization of the serine protease (caseinase) and metalloprotease (elastase). J Appl Microbiol 96: 944–1001.
- Falcón RM, Carvalho HF, Joazeiro PP, Gatti MS & Yano T (2001) Induction of apoptosis in HT-29 human intestinal epithelial cells by the cytotoxic enterotoxin of Aeromonas hydrophila. Biochem Cell Biol 79: 525–531.
- Fergunson MR, Xing-Jing Xu, Houston CW, Peterson JW, Coppenhaver DH & Chopra AK (1997) Hyperproduction, purification, and mechanics of actions of the cytotoxic enterotoxin produced by Aeromonas hydrophila. Infect Immunol 65: 4299–4308.
- Figueroa-Arredondo P, Heuser JE, Akopyants NS, Moriasaki JH, Cerezo SG, Rincón FE & Berg D (2001) Cell vacuolation caused by Vibrio colerae hemolysin. Infect Immunol 69: 1613–1624.
- Fujii Y, Nomura T, Kanzawa H, Kameyama M, Yamanaka H, Akita M, Setsu K & Okamoto K (1998) Purification and characterization of enterotoxin produced by Aeromonas sobria. Microbiol Immunol 42: 703–714.
- Fujii Y, Nomura T & Okamoto K (1999) Hemolysin of Aeromonas sobria stimulates production of cyclic AMP by cultured cells. FEMS Microbiol Lett 176: 67–72.
- Henics T & Wheatley DN (1999) Cytoplasmatic vacuolation, adaption and cell death: a view on new perspectives and features. Biology of the cell 91: 485–498.
- King GE, Werner SB & Kizer KW (1991) Epidemiology of Aeromonas infections in California. Clin Infect Dis 15: 449–452.
- Krovacek K, Faris A, Baloda SB, Peterz M, Lindberg T & Mansson I (1992) Prevalence and characterization of Aeromonas spp. isolated from foods in Uppsala, Sweden. Food Microbiol 9: 29–36.
- Laemmli UK (1970) Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
- Ljung A, Wretling B & Mollky R (1981) Separation and characterization of enterotoxin and two haemolysins from Aeromonas hydrophila. Acta Pathol Microbiol Scand 89: 387–397.
- Luzzi I, Covacci A, Censini S, Pezzella C, Crotti D & Facchini M (1996) Detection of a vacuolating cytotoxin in stools from children with diarrhea. Clin Infect Dis 23: 101–106.
- Martins LM, Falcón RM & Yano T (2002) Incidence of toxic Aeromonas isolated from food and clinical cases. FEMS Immunol Med Microbiol 32: 237–242.
- Mello MLS & Vidal BC (1980) Práticas de Biologia Celular. Edigard Blucher, Rio de Janeiro (Appendix 71).
- Mitchell DB, Santone KS & Acosta D (1980) Evaluation of cytotoxicity in cultured cells by enzyme leakage. J Tis Cult Methods 6: 113–116.
- Mitra R, Figueroa-Arredondo P, Mukopadhway AK, Shimada T, Takeda Y, Berg DE & Nair GB (2000) Cell vacuolation, a manifestation of the El Tor hemolysin of Vibrio cholerae. Infect Immunol 68: 1928–1933.
- Monfort P & Baleux B (1991) Haemolysin occurrence among Aeromonas hydrophila, Aeromonas caviae and Aeromonas sobria strains isolated from different aquatic ecosystems. Res Microbiol 142: 95–102.
- Rodriguez LA, Ellis AE & Nieto TP (1992) Purification and characterization of an extracellular metalloprotease, serine protease and haemolysin of Aeromonas hydrophila strain B32: all are letal for fish. Microbiol Pathogenesis 37: 17–24.
- Rose JM, Houston CW, Coppenhaver DH, Dixon JD & Kurosky K (1989) Purification and chemical characterization of a cholera toxin-cross-reactive cytolytic enterotoxin produced by a human isolate of Aeromonas hydrophila. Infect Immunol 57: 1170–1176.
- Song T, Toma C, Nakasone N & Iwanaga M (2004) Aerolysin is activated by metalloprotease in Aeromonas veronii biovar sobria. J Med Microbiol 53: 477–482.
- Trower CJ, Abo S, Majeed KN & Von Itzstein M (2000) Production of an enterotoxin by a gastro-enteritis-associated Aeromonas strain. J Med Microbiol 49: 121–126.
- Welkos S & O'Brien A (1994) Determination of median lethal and infectious doses in animal model systems. Methods Enzymol 235: 29–39.
- Yokoyama R, Fujji Y, Noguchi Y, Nomura T, Akita M, Setsu K, Yamamoto S & Okamoto K (2002) Physicochemical and biological properties of an extracellular serine protease of Aeromonas sobria. Microbiol Immunol 46: 383–390.
- Zang Z-Du & Gerard C (1996) MTT assay overestimates human airway smooth muscle cell number in culture. Biochem Mol Biol Intern 38: 431–436.
