Objective Secreted proteins of Helicobacter pylori （H. pylori） interact with gastric epithelium cells and may contribute to cell damage. Considering the fact that HPO17S and hypothetical conserved protein HP1286 are...Objective Secreted proteins of Helicobacter pylori （H. pylori） interact with gastric epithelium cells and may contribute to cell damage. Considering the fact that HPO17S and hypothetical conserved protein HP1286 are included in the group and that HP0175 is a well-known apoptosis-induced factor, the present study aims to clarify whether HP1286 plays a role in bacterial pathogenicity or even functions as an apoptosis-induced factor in human stomach. Methods Two genes encoding HP1286 and HPO175 were cloned into pET32a vector and expressed as recombinant proteins in Escherichia coil （E. coil） BL21. Signal peptide sequences were not included. The recombinant proteins were purified with His SpinTrap and desalted by using HiTrap Desalting. Immunoreactivity of the proteins was determined by Western blot. Human gastric epithelial cell AGS was challenged with these endotoxin-free proteins; and apoptosis of cells was assayed with the Cell Death ELISA kit. Results Recombinant proteins and their respective products whose N-terminal his-tag were removed with thrombin were recognized by serum from the patient infected with H. pylori. Apoptotic AGS cells challenged by HP1286 of H. pylori 26695 were four times more than untreated cells. In addition, apoptosis-induced ability of HP1286 of SS1 was not as strong as that of H. pylori 26695 strain. Conclusion HP1286 of H. pylori 26695 induces apoptosis of AGS cells in a time-dependent manner, however the apoptosis-induced ability of HP1286 may differ due to variations between different strains.展开更多
基金supported by the Major Technology Project as part of "Prevention and Control of Major Infectious Diseases including AIDS and Viral Hepatitis" (No. 2008ZX10004-002)
文摘Objective Secreted proteins of Helicobacter pylori （H. pylori） interact with gastric epithelium cells and may contribute to cell damage. Considering the fact that HPO17S and hypothetical conserved protein HP1286 are included in the group and that HP0175 is a well-known apoptosis-induced factor, the present study aims to clarify whether HP1286 plays a role in bacterial pathogenicity or even functions as an apoptosis-induced factor in human stomach. Methods Two genes encoding HP1286 and HPO175 were cloned into pET32a vector and expressed as recombinant proteins in Escherichia coil （E. coil） BL21. Signal peptide sequences were not included. The recombinant proteins were purified with His SpinTrap and desalted by using HiTrap Desalting. Immunoreactivity of the proteins was determined by Western blot. Human gastric epithelial cell AGS was challenged with these endotoxin-free proteins; and apoptosis of cells was assayed with the Cell Death ELISA kit. Results Recombinant proteins and their respective products whose N-terminal his-tag were removed with thrombin were recognized by serum from the patient infected with H. pylori. Apoptotic AGS cells challenged by HP1286 of H. pylori 26695 were four times more than untreated cells. In addition, apoptosis-induced ability of HP1286 of SS1 was not as strong as that of H. pylori 26695 strain. Conclusion HP1286 of H. pylori 26695 induces apoptosis of AGS cells in a time-dependent manner, however the apoptosis-induced ability of HP1286 may differ due to variations between different strains.
