Ceponis Peter J M, McKay Derek M, Menaker Rena J, Galindo-Mata Esther, Jones Nicola L
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
J Immunol. 2003 Aug 15;171(4):2035-41. doi: 10.4049/jimmunol.171.4.2035.
Helicobacter pylori is a bacterial pathogen evolved to chronically colonize the gastric epithelium, evade immune clearance by the host, and cause gastritis, peptic ulcers, and even gastric malignancies in some infected humans. In view of the known ability of this bacterium to manipulate gastric epithelial cell signal transduction cascades, we determined the effects of H. pylori infection on epithelial IL-4-Stat6 signal transduction. HEp-2 and MKN45 epithelial cells were infected with H. pylori strains LC11 or 8823 (type 1; cagA(+)/cagE(+)/VacA(+)), LC20 (type 2; cagA(-), cagE(-), VacA(-)), and cagA, cagE, and vacA isogenic mutants of strain 8823, with some cells receiving subsequent treatment with the Th2 cytokine IL-4, a known Stat6 activator. Immunofluorescence showed a disruption of Stat6-induced nuclear translocation by IL-4 in LC11-infected HEp-2 cells. IL-4-inducible Stat6 DNA binding in HEp-2 and MKN45 cells was abrogated by infection, but MKN45 cell viability was unaffected. A decrease in IL-4-mediated Stat6 tyrosine phosphorylation in nuclear and whole cell lysates was also observed following infection with strains LC11 and LC20, while neither strain altered IL-4 receptor chain alpha or Janus kinase 1 protein expression. Furthermore, parental strain 8823 and its isogenic cagA, cagE, and vacA mutants also suppressed IL-4-induced Stat6 tyrosine phosphorylation to comparable degrees. Thus, H. pylori did not directly activate Stat6, but blocked the IL-4-induced activation of epithelial Stat6. This may represent an evolutionarily conserved strategy to disrupt a Th2 response and evade the host immune system, allowing for successful chronic infection.
幽门螺杆菌是一种细菌病原体,它能够长期定殖于胃上皮细胞,逃避宿主的免疫清除,并在部分感染者中引发胃炎、消化性溃疡,甚至胃癌。鉴于该细菌已知具有操纵胃上皮细胞信号转导级联反应的能力,我们测定了幽门螺杆菌感染对上皮白细胞介素-4-信号转导子和转录激活子6(IL-4-Stat6)信号转导的影响。用幽门螺杆菌菌株LC11或8823(1型;细胞毒素相关基因A阳性(cagA(+))/细胞毒素相关基因E阳性(cagE(+))/空泡毒素A阳性(VacA(+)))、LC20(2型;cagA(-)、cagE(-)、VacA(-))以及8823菌株的cagA、cagE和vacA等基因缺失突变体感染人喉表皮样癌细胞(HEp-2)和人胃癌细胞(MKN45),部分细胞随后用Th2细胞因子IL-4(一种已知的Stat6激活剂)进行处理。免疫荧光显示,在感染了LC11的HEp-2细胞中,IL-4诱导的Stat6核转位受到破坏。感染使HEp-2和MKN45细胞中IL-4诱导的Stat6与DNA结合能力丧失,但MKN45细胞活力未受影响。在用菌株LC11和LC20感染后,还观察到核裂解物和全细胞裂解物中IL-4介导的Stat6酪氨酸磷酸化水平降低,而这两种菌株均未改变IL-4受体α链或Janus激酶1蛋白的表达。此外,亲本菌株8823及其cagA、cagE和vacA等基因缺失突变体也能以相当的程度抑制IL-4诱导的Stat6酪氨酸磷酸化。因此,幽门螺杆菌并非直接激活Stat6,而是阻断了IL-4诱导的上皮细胞Stat6激活。这可能代表了一种进化上保守的策略,用于破坏Th2反应并逃避宿主免疫系统,从而实现成功的慢性感染。