Malik-Kale Preeti, Parker Craig T, Konkel Michael E
School of Molecular Biosciences, Washington State University, Pullman, WA 99164, USA.
J Bacteriol. 2008 Apr;190(7):2286-97. doi: 10.1128/JB.01736-07. Epub 2008 Jan 25.
Campylobacter jejuni, a spiral-shaped gram-negative bacterium, is a leading bacterial cause of human food-borne illness. Acute disease is associated with C. jejuni invasion of the intestinal epithelium. Further, maximal host cell invasion requires the secretion of proteins termed Campylobacter invasion antigens (Cia). As bile acids are known to alter the pathogenic behavior of other gastrointestinal pathogens, we hypothesized that the virulence potential of Campylobacter may be triggered by the bile acid deoxycholate (DOC). In support of this hypothesis, culturing C. jejuni with a physiologically relevant concentration of DOC significantly altered the kinetics of cell invasion, as shown by gentamicin protection assays. In contrast to C. jejuni harvested from Mueller-Hinton (MH) agar plates, C. jejuni harvested from MH agar plates supplemented with DOC secreted the Cia proteins, as judged by metabolic labeling experiments. DOC was also found to induce the expression of the ciaB gene, as determined by beta-galactosidase reporter, real-time reverse transcription-PCR, and microarray analyses. Microarray analysis further revealed that DOC induced the expression of virulence genes (ciaB, cmeABC, dccR, and tlyA). In summary, we demonstrated that it is possible to enhance the pathogenic behavior of C. jejuni by modifying the culture conditions. These results provide a foundation for identifying genes expressed by C. jejuni in response to in vivo-like culture conditions.
空肠弯曲菌是一种螺旋形革兰氏阴性菌,是人类食源性疾病的主要细菌病因。急性疾病与空肠弯曲菌侵袭肠上皮细胞有关。此外,最大程度的宿主细胞侵袭需要分泌一种名为弯曲菌侵袭抗原(Cia)的蛋白质。由于已知胆汁酸会改变其他胃肠道病原体的致病行为,我们推测空肠弯曲菌的毒力可能由胆汁酸脱氧胆酸盐(DOC)触发。为支持这一假设,用生理相关浓度的DOC培养空肠弯曲菌显著改变了细胞侵袭动力学,庆大霉素保护试验表明了这一点。与从穆勒-欣顿(MH)琼脂平板收获的空肠弯曲菌相比,从添加了DOC的MH琼脂平板收获的空肠弯曲菌分泌了Cia蛋白,代谢标记实验判断得出。通过β-半乳糖苷酶报告基因、实时逆转录PCR和微阵列分析确定,DOC还能诱导ciaB基因的表达。微阵列分析进一步显示,DOC诱导了毒力基因(ciaB、cmeABC、dccR和tlyA)的表达。总之,我们证明通过改变培养条件有可能增强空肠弯曲菌的致病行为。这些结果为鉴定空肠弯曲菌在类似体内培养条件下表达的基因奠定了基础。