تعيين الگوي آنتي ژني هليكوباكترپيلوري جدا شده از بيماران مبتلا به اولسر پپتيك به روش ايمنوبلاتينگ

Pattern of Helicobacter pylori Antigens isolated from Patients with Peptic Ulcer by immunoblotting

helicobacter pylori is an important etiologic cause of chronic infection of gastric mucus, chronic gastritis, peptic ulcers and gastric cancer in human. The aim of this study was to identify and characterize the dominant antigen of H.pylori, which is responsible for the humoral and cellular immune responses. Gastric biopsy of patients with gastric ulcers were sent to microbiology lab. First, samples were homogenized at sterile conditions, and then they were cultured in special medium and micro-aerophylic conditions. 25 colonies of h.pylori were removed from culture medium and the whole-cell lysates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunoblots were performed using sera from h.pyloriinfected persons . Specific h.pylori antibody responses in persons were analyzed by enzyme-linked immunosorbent assay. Two groups of bands appeared after staining of the SDS-PAGE with comassie blue. One group was within 55-97KD and the other were around 20-24 KD. Western - blotting analysis detected a band around 55-97 KD, which is dominant antigen and responsible for immune response against h.pylori. Using ammonium sulfate as a precipitant for sera would have valuable effects in the westernblot results. As it is shown in the results , an antigen with MW=97 KD is immunodominant and stimulate patientʹs immune system to produce antibody and can be candidate as a subunit vaccine in future.