Wagner S, Beil W, Mai U E, Bokemeyer C, Meyer H J, Manns M P
Department of Gastroenterology, and Hepatology, Medizinische Hochschule Hannover, FRG.
Pharmacology. 1994 Oct;49(4):226-37. doi: 10.1159/000139238.
A human in vitro model to study the interaction between Helicobacter pylori and gastric epithelial cells was developed using primary cultures of gastric mucosal cells (isolated from gastric biopsies or operative specimen and maintained in culture for 2 weeks) as well as the well-differentiated human gastric carcinoma cell line HM02, the undifferentiated gastric tumour cell line HM51, and the laryngeal epithelial cell line HEp-2. Primary cultures and all cell lines were exposed to seven isolates of H. pylori isolated from gastritis and duodenal ulcer patients. Microbial adherence was assessed by microscopical evaluation of Giemsa-stained preparations and by culturing the viable bacteria attached to the epithelial cells. All H. pylori isolates adhered to the gastric cells in primary culture, to HM02 cells, and to HEp-2 cells with the greatest binding affinity found in primary gastric cells. No adherence was detected in HM51 cells. H. pylori adherence was dependent on bacterial load, incubation time, and temperature. There was no difference in microbial binding between H. pylori isolates derived from gastritis and duodenal ulcer patients. The effect of antiulcer drugs on H. pylori adherence was investigated by pre-incubating isolates of H. pylori with omeprazole, cimetidine, and bismuth subcitrate. Omeprazole and cimetidine failed to significantly influence microbial adherence. In contrast, bismuth subcitrate already in concentrations below the MIC range decreased H. pylori adherence in gastric epithelial cells and in HEp-2 cells substantially. Our study shows that primary cultured human gastric mucosal cells and the human gastric carcinoma cell line HM02 provide suitable in vitro models for the study of the interactions between H. pylori and the gastric epithelium. This gastric cell model is characterized by a high affinity for H. pylori binding.
利用胃黏膜细胞原代培养物(从胃活检组织或手术标本中分离并在培养中维持2周)以及高分化人胃癌细胞系HM02、未分化胃肿瘤细胞系HM51和喉上皮细胞系HEp-2,建立了一种用于研究幽门螺杆菌与胃上皮细胞相互作用的体外模型。原代培养物和所有细胞系均暴露于从胃炎和十二指肠溃疡患者中分离出的7株幽门螺杆菌。通过对吉姆萨染色制剂进行显微镜评估以及培养附着在上皮细胞上的活菌来评估微生物黏附情况。所有幽门螺杆菌分离株均黏附于原代培养的胃细胞、HM02细胞和HEp-2细胞,其中在原代胃细胞中发现的结合亲和力最高。在HM51细胞中未检测到黏附。幽门螺杆菌的黏附取决于细菌载量、孵育时间和温度。源自胃炎和十二指肠溃疡患者的幽门螺杆菌分离株之间的微生物结合没有差异。通过将幽门螺杆菌分离株与奥美拉唑、西咪替丁和枸橼酸铋预先孵育,研究了抗溃疡药物对幽门螺杆菌黏附的影响。奥美拉唑和西咪替丁未能显著影响微生物黏附。相比之下,浓度低于最低抑菌浓度范围的枸橼酸铋就可显著降低幽门螺杆菌在胃上皮细胞和HEp-2细胞中的黏附。我们的研究表明,原代培养的人胃黏膜细胞和人胃癌细胞系HM02为研究幽门螺杆菌与胃上皮之间的相互作用提供了合适的体外模型。这种胃细胞模型的特点是对幽门螺杆菌结合具有高亲和力。