Cytotoxic isolates of Helicobacter pylori from peptic ulcer diseases decrease K+-dependent ATPase activity in HeLa cells.
BACKGROUND
Helicobacter pylori is a Gram negative bacterium that plays a central role in the etiology of chronic gastritis and peptic ulcer diseases. However, not all H. pylori positive cases develop advanced disease. This discriminatory behavior has been attributed to the difference in virulence of the bacteria. Among all virulence factors, cytotoxin released by H. pylori is the most important factor. In this work, we studied variation in H. pylori isolates from Indian dyspeptic patients on the basis of cytotoxin production and associated changes in K+-dependent ATPase (one of its targets) enzyme activity in HeLa cells.


METHODS
The patients were retrospectively grouped on the basis of endoscopic and histopathological observation as having gastritis or peptic ulcer. The HeLa cells were incubated with the broth culture filtrates (BCFs) of H. pylori isolates from patients of both groups and observed for the cytopathic effects: morphological changes and viability. In addition, the K+-dependent ATPase activity was measured in HeLa cells extracts.


RESULTS
The cytotoxin production was observed in 3/7 (gastritis) and 4/4 (peptic ulcer) H. pylori isolates. The BCFs of cytotoxin producing H. pylori strains reduced the ATPase activity of HeLa cells to 40% of that measured with non-cytotoxin producing H. pylori strains (1.33 micromole Pi/mg protein and 3.36 micromole Pi/mg protein, respectively, p < 0.05). The decreased activity of ATPase enzyme or the release of cytotoxin also correlated with the increased pathogenicity indices of the patients.


CONCLUSIONS
Our results suggest that the isolation of cytotoxic H. pylori is more common in severe form of acid peptic diseases (peptic ulcer) than in gastritis patients from India. Also the cytotoxin released by H. pylori impairs the ion-transporting ATPase and is a measure of cytotoxicity.
Shanjana
Awasthi
A
Department of Microbiology, Sanjay Gandhi P,G, Institute of Medical Sciences, Lucknow, UP, India. awasthis@uthscsa.edu
Archana
Ayyagari
A
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000293:Adolescent; D000328:Adult; D001706:Biopsy; D027682:Cation Transport Proteins; D002648:Child; D003603:Cytotoxins / Q000378:metabolism*; D005756:Gastritis / Q000382:microbiology*; D006367:HeLa Cells / Q000201:enzymology; D016481:Helicobacter Infections / Q000382:microbiology*; D016480:Helicobacter pylori / Q000302:isolation & purification / Q000378:metabolism / Q000472:pathogenicity*; D006801:Humans; D011706:Pyloric Antrum / Q000382:microbiology / Q000648:ultrastructure; D012189:Retrospective Studies; D013276:Stomach Ulcer / Q000382:microbiology*; D014774:Virulence
D027682:Cation Transport Proteins; D003603:Cytotoxins; D000251:Adenosine Triphosphatases; C022761:potassium transporting ATPase