Crabtree J E, Farmery S M, Lindley I J, Figura N, Peichl P, Tompkins D S
Department of Clinical Medicine, St James's University Hospital, Leeds.
J Clin Pathol. 1994 Oct;47(10):945-50. doi: 10.1136/jcp.47.10.945.
To investigate: (1) whether Helicobacter pylori directly induces interleukin-8 (IL-8) message expression and protein secretion in established gastric epithelial cell lines; and (2) if CagA/cytotoxin positive and negative strains of H pylori differ in their ability to induce epithelial IL-8.
Gastric epithelial cell lines were co-cultured with H pylori NCTC 11637 and 10 clinical isolates (four cytotoxic, six non-cytotoxic) and secreted IL-8 was measured by enzyme linked immunosorbent assay (ELISA). Specific induction of gastric epithelial IL-8 mRNA was examined by reverse transcription and polymerase chain reaction (RT-PCR) amplification.
H pylori (NCTC 11637) induced IL-8 secretion from three gastric epithelial cell lines (KATO-3, ST42, AGS) but not from MKN 45 (gastric) or intestinal (SW480, HT29) cell lines. H mustelae did not stimulate IL-8 secretion from KATO-3, ST42, and AGS cells. H pylori induced IL-8 secretion was reduced by heat killing, sonication, freeze thawing or formalin fixation of the bacteria. CagA/cytotoxin positive strains of H pylori induced significantly higher IL-8 secretion than CagA/cytotoxin negative strains in the three positive gastric epithelial cell lines (KATO-3, ST42: p < 0.01; AGS: p < 0.02). A significant increase (p < 0.01) in the expression of IL-8 mRNA relative to G3PDH mRNA was observed in KATO-3 cells after three hours of co-culture with CagA/cytotoxin positive strains.
H pylori directly increases gastric epithelial IL-8 mRNA expression and IL-8 protein secretion in a strain specific manner. Induction of epithelial IL-8 by CagA/cytotoxin positive strains is likely to result in neutrophil chemotaxis and activation and thus mucosal damage. These observations on epithelial IL-8 may explain the association between CagA/cytotoxin positive strains and gastroduodenal disease.
研究：（1）幽门螺杆菌是否直接诱导已建立的胃上皮细胞系中白细胞介素-8（IL-8）的信使表达和蛋白质分泌；以及（2）幽门螺杆菌的细胞毒素相关基因A（CagA）/细胞毒素阳性和阴性菌株在诱导上皮细胞IL-8方面的能力是否存在差异。
将胃上皮细胞系与幽门螺杆菌NCTC 11637及10株临床分离株（4株具有细胞毒性，6株无细胞毒性）共培养，采用酶联免疫吸附测定（ELISA）法检测分泌的IL-8。通过逆转录和聚合酶链反应（RT-PCR）扩增检测胃上皮IL-8 mRNA的特异性诱导情况。
幽门螺杆菌（NCTC 11637）诱导3种胃上皮细胞系（KATO-3、ST42、AGS）分泌IL-8，但未诱导MKN 45（胃）或肠（SW480、HT29）细胞系分泌。幽门弯曲菌未刺激KATO-3、ST42和AGS细胞分泌IL-8。幽门螺杆菌经热灭活、超声处理、冻融或福尔马林固定后，诱导的IL-8分泌减少。在3种阳性胃上皮细胞系（KATO-3、ST42：p < 0.01；AGS：p < 0.02）中，幽门螺杆菌CagA/细胞毒素阳性菌株诱导的IL-8分泌显著高于CagA/细胞毒素阴性菌株。与CagA/细胞毒素阳性菌株共培养3小时后，KATO-3细胞中相对于甘油醛-3-磷酸脱氢酶（G3PDH）mRNA，IL-8 mRNA表达显著增加（p < 0.01）。
幽门螺杆菌以菌株特异性方式直接增加胃上皮IL-8 mRNA表达和IL-8蛋白质分泌。CagA/细胞毒素阳性菌株诱导上皮细胞IL-8可能导致中性粒细胞趋化和活化，进而造成黏膜损伤。这些关于上皮细胞IL-8的观察结果可能解释了CagA/细胞毒素阳性菌株与胃十二指肠疾病之间的关联。
