TITLE

PCR DETECTION OF CAG PATHOGENICITY ISLAND GENES DOES NOT ACCURATELY PREDICT FUNCTIONALITY OF CAG ENCODED PROTEINS

AUTHOR(S)
Argent, R.H.; Kidd, M.; Owen, R.J.; Limb, M.C.; Atherton, J.C.
PUB. DATE
April 2003
SOURCE
Gut;Apr2003 Supplement 1, Vol. 52, pA17
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
H pylori associated disease is more common in patients infected with CagA+ strains. CagA is one of 31 proteins encoded on the cag pathogenicity island (Pal). CagA itself is translocated into host cells via a syringe like structure comprising other cag encoded proteins. Here it becomes tyrosine phosphorylated and causes proliferative cell signalling and cytoskeletal changes. Interaction of the cag syringe itself with epithelial cells results in induction of proinflammatory cytokines, particularly interleukin-8 (IL-8). One or both of cagA phosphorylation and cag induced IL-8 secretion are likely responsible for the increased disease risk from CagA+ strains. Thus, we aimed to assess whether PCR detection of selected cag Pal genes could predict whether H pylori strains could induce epithelial cells to phosphorylate cagA and/or secrete IL-8. Methods/Results: H pylori strains were obtained from 26 patients from South Africa. The presence of cagA, cagE, cagM, cagT, and cag6-7 genes was determined by PCR analysis, cagA was present in all 26 strains, and 16 had all the tested cag Pal genes and so were predicted to encode an intact Pal. Possession and phosphorylation of cagA was determined by use of a cell free system and by co-culturing H pylori with AGS cells at 37° for 6 h, followed by lysis of the cells, and western blotting. Of the 26 strains, 20 induced cagA phosphorylation within AGS cells, and all of these strains induced the secretion of IL-8. Of the remaining 6 strains none induced IL-8 secretion. The degree of cagA phosphorylation induced and the amount of translocated cagA were not related to the level of lL-8 secretion induced. Six strains that possessed cagA did not produce cagA protein. Four strains predicted to lack a complete cag Pal by PCR analysis induced both cagA phosphorylation and IL-8 production, and one strain predicted to possess an intact Pal by PCR did not induce cagA phosphorylation or IL-8 production. Conclusion: PCR analysis of selected...
ACCESSION #
9747370

 

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