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MOI-Dependent Effects of cagA+ and cagA- H. pylori on IL-8 Promoter Activity in AGS Cells

This study investigates the influence of cagA+ and cagA- Helicobacter pylori strains on NF-kB binding site activity in the IL-8 promoter using AGS cells. Cells were transfected with luciferase reporter plasmids containing either the wild-type IL-8 promoter or a NF-kB mutant variant. Following infection with different H. pylori strains and multiplicities of infection (MOI), luciferase assays were conducted to measure the promoter activity. Significant differences in induction were observed, highlighting the role of cagA status in IL-8 transcriptional regulation.

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MOI-Dependent Effects of cagA+ and cagA- H. pylori on IL-8 Promoter Activity in AGS Cells

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  1. Fold Induction 10X 100X 10X 100X 10X 100X control HP87 G27 G27-delcagA ** Supplementary Figure 2. MOI-dependent effects of cagA+ and cagA-H. pylori strains on NF-kB binding site in IL-8 promoter. AGS cells were transfected with 1µg Renillaluciferase expression plasmid and 5µg of reporter plasmids containing firefly luciferase coding sequences derived by wild type IL-8 promoter (wt, black bars) or NFkB mutant promoter (nfkb, white bars). IL-8 wild type and mutant promoter sequences are shown at the top. After 48 hours cells were infected with different H. pylori strains and MOIs as indicated. After 18 hours, cells were harvested and cellular extracts were prepared and subjected to luciferase assay. Firefly luciferase light units were normalized to Renillaluciferase activity to obtain the relative firefly light units of each reporter plasmid. Relative firefly light units in non infected cells (control) in a single experiment were set to 1 to obtain the induction factor for wild type and mutant reporters in H. pylori infected cells (Fold induction). Mean values of fold induction are shown +/- S.E.M. from three independent transfection experiments in AGS cells. The symbol ** indicates significant difference (P < 0.01) between cells transfected with wild type IL-8 promoter or NFkB mutant promoter.

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