Production of specific IgY Helicobacter pylori recombinant OipA protein and assessment of its inhibitory effects towards attachment of H. pylori to AGS cell line

PURPOSE: The common triple therapy for Helicobacter pylori is challenged by the increasing cases of antibiotic resistant infections, raising the need to explore alternative therapies. Oral administration of egg yolk immunoglobulin Y (IgY) has been previously reported as a means of passive immunization therapy for H. pylori infections. In this work, we investigated the inhibitory effect of IgY on the attachment of H. pylori to AGS cell line. MATERIALS AND METHODS: Recombinant OipA was prepared. Hens were immunized with recombinant protein three times. IgY was purified from egg yolks of immunized hens using polyethylene glycol precipitation method. The inhibitory effect of the specific immunoglobulin was evaluated in AGS cell line infected with H. pylori. RESULTS: The presence of recombinant OipA (30 kD) was confirmed via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunization of hens was confirmed using enzyme-linked immunosorbent assay. The purified IgY from egg yolks were assessed using SDS-PAGE and confirmed by western blot. CONCLUSION: The results showed that IgY-OipA had inhibitory effect on attachment of H. pylori to AGS cell line and may be utilized as a therapeutic or prophylaxis material.

[Cytotoxicity effect of recombinant outer membrane inflammatory protein [oipA] of helicobacter pylori on a breast cancer cell line]

Breast cancer is one of the leading causes of death in women worldwide. Conventional treatments use cytotoxic drugs which have high numbers of side effects. Currently pharmacologists are searching for novel drugs with fewer side effects and maximum efficiency as breast cancer treatment. The aim of the current study is to clarify the cytotoxicity effect of the recombinant outer membrane inflammatory protein [oipA] of Helicobacter pylori [H. pylori] on a breast cancer cell line. We purified recombinant H. pylori oipA by Ni-NTA affinity chromatography. Breast cancer cells [4T1] were treated with different concentrations of recombinant oipA for various lengths of time. Cell viability was evaluated by the viability assay [MTT test]. SDS-PAGE analysis showed the expression of an approximately 34000 dalton protein. Statistical analysis showed oipA toxic effects on 4T1 cells at a concentration of 250 micro g/ml after 24 h. These findings suggested that oipA had a direct toxic effect on a breast cancer cell line [4T1] in vitro. The oipA protein might be a new tool for future therapeutic strategies in cancer immunotherapy

beta-lactamase typing by substrate hydrolysis in clinical isolates of methicillin resistant Staphylococcus aureus

Staphylococcus aureus is one of the most important causes of nosocomial infections and most clinical isolates are multidrug resistant. Resistance to beta-lactam antibiotics is most often due to bacterial beta-lactamase production. Characterization of beta-lactamases is important for choosing appropriate antibiotic therapy. Thirty methicillin resistant Staphylococcus aureus [MRSA] were identified by standard biochemical methods. Antibacterial susceptibility to 9 beta-lactam antibiotics was determined. Beta-lactamase production was shown in all isolates using the colony iodometric test and nitrocefin discs. Beta-lactamase typing was carried out by measuring the relative substrate hydrolysis rates. The MRSA isolates were resistant to the majority of beta-lactam antibiotics. The results showed that 90% of the isolates displayed type A substrate hydrolysis profile of beta-lactamase. The alarming high level of resistance to beta-lactam antibiotics including methicillin and 3[rd] generation beta-lactams show the need for extensive studies on alternative treatment protocols and use of new drugs