Helicobacter pylori Vacuolating Toxin Exhibits Polar Activity of $Cl^-$ Secretion and Secretory Response to Carbachol in T84 Cells

  • Jin, Nan-Ge (Department of Physiology and Biophysics, Seoul Notional University College of Medicine) ;
  • Jin, Yong-Ri (Department of Digestive medicine, Affiliated Hospital of Yanbian University College of Medicine) ;
  • So, In-Suk (Department of Physiology and Biophysics, Seoul Notional University College of Medicine) ;
  • Kim, Ki-Whan (Department of Physiology and Biophysics, Seoul Notional University College of Medicine)
  • Published : 2004.10.21

Abstract

To investigate whether VacA (vacuolating toxin) produced by Helicobacter pylori Korean stain 99 induces intestinal secretion, purified VacA was added to T84 cell monolayers mounted in Ussing chambers, and electrical parameters were monitored. Mucosal addition of low pH-pretreated VacA increased short circuit current (Isc). The effect was time- and dose-dependent and saturable. The time-to-peak Isc was concentration-dependent. Chloride channel inhibitors, niflumic acid or 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), inhibited VacA-stimulated Isc. Carbachol (CCh)-induced increase of Isc was prolonged by the addition of VacA to the mucosal side only. The effect was unaltered by the addition of niflumic acid. VacA did not show cytopathic effects. These studies indicate that VacA is a nonlethal toxin that acts in a polar manner on T84 monolayers to potentiate $Cl^-$ secretion and the response to CCh secretion without decrease in monolayer resistance. VacA may contribute to diarrhea diseases in human intestinal epithelial cells.

Keywords

References

  1. Atherton JC, Peek RMJ, Tham KT, Cover TL, Blaser MJ. Clinical and pathological importance of heterogeneity in VacA, the vacuolating cytotoxin gene of Helicobacter pylori. Gastroenterology 112: 92-99, 1997 https://doi.org/10.1016/S0016-5085(97)70223-3
  2. Blaser MJ. Helicobacter pylori: microbiology of a "slow" bacterial infection. Trends Microbiol 1: 255-260, 1993 https://doi.org/10.1016/0966-842X(93)90047-U
  3. Cover TL, Cao P, Lind CD, Tham KT, Blaser MJ. Correlation between vacuolating cytotoxin production by Helicobacter pylori isolates in vitro and in vivo. Infect Immun 61: 5008-5012, 1993
  4. Dharmsathaphorn K, Pandol SJ. Mechanisms of chloride secretion induced by carbachol in a colonic epithelial cell line. J Clin Invest 77: 348-354, 1986 https://doi.org/10.1172/JCI112311
  5. Dorrell L, Wardropper AG, Ong ELC. Helicobacter pylori infection in an HIV-seropositive patient with diarrhoea. AIDS 7: 900- 901, 1993 https://doi.org/10.1097/00002030-199306000-00028
  6. Grasset E, Bernabeu J, Pinto M. Epithelial properties of human colonic carcinoma cell line Caco-2: effect of secretagogues. Am J Physiol 248: C410-418, 1985 https://doi.org/10.1152/ajpcell.1985.248.5.C410
  7. Guarino A, Bisceglia M, Canani RB, Boccia MC, Mallardo G, Bruzzese E, Massari P, Rappuoli R, Telford J. Enterotoxic effect of the vacuolating toxin produced by Helicobacter pylori in Caco-2 cells. J Infect Dis 178: 1373-1378, 1998 https://doi.org/10.1086/314427
  8. Guarino A, Canani RB, Casola A, Pozio E, Russo R, Bruzzese E, Fontana M, Rubino A. Human intestinal cryptosporidiosis: secretory diarrhea and enterotoxic activity in Caco-2 cells. J Infect Dis 171: 976-983, 1995
  9. Guarino A, Casola A, Bruzzese E, Saini M, Nitsch L, Rubino A. Human serum immunoglobulin counteracts rotaviral infection in Caco-2 cells. Pediatr Res 40: 881-887, 1996 https://doi.org/10.1203/00006450-199612000-00019
  10. Luzzi I, Covacci A, Censini S, Pezzella C, Crotti D, Facchini M, Giammanco A, Guglielmetti P, Piersimoni C, Bonamico M, Mariani P, Rappuoli R, Caprioli A. Detection of a vacuolating cytotoxin in stools from children with diarrhea. Clin Infect Dis 23: 101-106, 1996 https://doi.org/10.1093/clinids/23.1.101
  11. Luzzi I, Pezzella C, Caprioli A, Covacci A, Bugnoli M, Censini S. Detection of vacuolating toxin of Helicobacter pylori in human faeces [letter]. Lancet 341: 1348, 1993
  12. Manetti R, Massari P, Burroni D, de Bernard M, Marchini A, Olivieri R, Papini E, Montecucco C, Rappuoli R, Telford JL. Helicobacter pylori cytotoxin: importance of native conformation for the induction of neutralizing antibodies. Infect Immun 63: 4476-4480, 1995
  13. Marchetti M, Arico B, Burroni D, Figura N, Rappuoli R, Ghiara P. Development of a mouse model of Helicobacter pylori infection that mimics human disease. Science 265: 1656-1658, 1995 https://doi.org/10.1126/science.7521969
  14. Matthews JB, Tally KJ, Smith JA, Zeind AJ, Hrnjez BJ. Activation of Cl secretion during chemical hypoxia by endogenous release of adenosine in intestinal epithelial monolayers. J Clin Invest 96: 117-125, 1995 https://doi.org/10.1172/JCI118010
  15. Papini E, Bugnoli M, De Bernard M, Figura N, Rappuoli R, Montecucco C. Bafilomycin A1 inhibits Helicobacter pylori induced vacuolization of HeLa cells. Mol Microbiol 7: 323-327, 1995
  16. Patel P, Mendall MA, Khulusi S, Northfield TC, Strachan DP. Helicobacter pylori infection in childhood: risk factors and effect on growth. Br Med J 309: 1119-1123, 1994 https://doi.org/10.1136/bmj.309.6962.1119
  17. Sullivan PB, Thomas JE, Wight DGD, Neale G, Eastham EJ, Corrah T, Lloyd-Evans N, Greenwood BM. Helicobacter pylori in Gambian children with chronic diarrhoea and malnutrition. Arch Dis Child 65: 189-1891, 1990 https://doi.org/10.1136/adc.65.2.189
  18. Szabo I, Brutsche S, Tombola F, Moschioni M, Satin B, Telford JL, Rappuoli R, Montecucco C, Papini E, Zoratti M. Formation of anion-selective channels in the cell plasma membrane by the toxin VacA of Helicobacter pylori is required for its biological activity. EMBO 18: 5517-5527,1999 https://doi.org/10.1093/emboj/18.20.5517
  19. Telford JL, Ghiara P, Dell'Orco M, Comanducci M, Burroni D, Bugnoli M, Tecce MF, Censini S, Covacci A, Xiang Z, Papini E, Montecucco C, Parente L, Rappuoli R. Gene structure of the Helicobacter pylori cytotoxin and evidence of its key role in gastric disease. J Exp Med 179: 1653-1658, 1994 https://doi.org/10.1084/jem.179.5.1653
  20. Thomas J, Gibson G, Darboe M, Dale A, Weaver L. Isolation of Helicobacter pylori from human faeces. Lancet 340: 1194- 11195, 1992 https://doi.org/10.1016/0140-6736(92)92894-L
  21. Xiang Z, Censini S, Bayeli PF, Telford JL, Figura N, Rappuoli R, Covacci A. Analysis of expression of CagA and VacA virulence factors in 43 strains of Helicobacter pylori reveals that clinical isolates can be divided into two major types and that CagA is not necessary for expression of the vacuolating cytotoxin. Infect Immun 63: 94-98, 1995