The adhesion of Helicobacter pylori extract to four mammalian cell lines.
OBJECTIVE
To study the adherence of acid-glycine extract (AGE) from Helicobacter pylori to four mammalian cell lines: KATO III, CCL17, CCL156 and Neuro-2A.


METHODS
In vitro assays to assess H. pylori adherence were based on the principle of the affinity of the bacterial antigens to mammalian cells in culture. Protein extracts from mammalian cells were either coated onto 96-well microtitre plates or electrophoretically resolved and blotted onto immobilon-P membranes. When the adhesive proteins derived from the AGE of H. pylori adhered to the mammalian cell proteins, these adhesive antigens were detected by rabbit anti-H. pylori AGE antibodies.


METHODS
The adhesive proteins in H. pylori NCTC11637 AGE were detected by enzyme-linked immunosorbent assay (ELISA) and Western blot under non-denaturing conditions.


RESULTS
Adhesive proteins constitute 5% H. pylori AGE proteins. The adhesive proteins adhered more readily to mammalian cell extracts than to bovine serum albumin. Modified Western blot analysis showed their affinity to a number of mammalian proteins ranging in molecular weight from 40 to 900 kDa, and in particular to the 140-230 kDa proteins.


CONCLUSIONS
The two in vitro adherence assays used effectively demonstrated the adherence of H. pylori to different mammalian cell lines. The 5% adhesive proteins from H. pylori may play an essential role in infection and colonization by H. pylori. Furthermore, adhesive proteins have similar adherence properties to mammalian cell components and may play an important role in the pathogenesis of H. pylori infection.
Ho
B
B
Department of Microbiology, National University of Singapore.
Jiang
B
B
D018829:Adhesins, Bacterial; D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology*; D001426:Bacterial Proteins / Q000032:analysis*; D015153:Blotting, Western; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D016480:Helicobacter pylori / Q000737:chemistry*; D006801:Humans; D051379:Mice
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; C073899:adhesin, helicobacter