Hanawa T, Yamamoto T, Kamiya S
Department of Microbiology, Kyorin University School of Medicine, Tokyo, Japan.
Infect Immun. 1995 Dec;63(12):4595-9. doi: 10.1128/iai.63.12.4595-4599.1995.
Listeria monocytogenes is a facultative intracellular pathogen which is able to survive and grow within phagocytic cells. Some facultative intracellular bacteria have been shown to respond to the hostile environment within phagocytic cells by producing a set of stress proteins. Since L. monocytogenes has a mechanism for intracellular survival that is distinct from those of other bacteria, we studied the phenotypic response of the bacterium to phagocytosis by macrophages. After phagocytosis of L. monocytogenes EGD by J774-1 macrophage cells, the microorganism rapidly increased in numbers about 20-fold during an incubation period of 5 h. In this phase of phagocytosis, the selective induction of 32 proteins was observed by two-dimensional gel electrophoresis. The responses to the environmental stresses of heat and hydrogen peroxide were also studied, and it was found that 14 heat shock proteins and 13 oxidative stress proteins were induced. Five of the induced proteins were common to both heat and oxidative stresses. By amino acid sequencing analysis, homologs of DnaK and GroEL were confirmed among the heat shock proteins. A comparison of the autoradiograms of the two-dimensional gels revealed that none of these stress proteins were among the proteins induced by L. monocytogenes within the macrophages. This behavior is entirely different from that shown by other facultative intracellular pathogens. Stress proteins known to be induced by environmental stresses were absent in intracellularly grown L. monocytogenes in the present study. This absence could be due to the mechanism by which the microorganisms rapidly escape from this stressful environment at a very early phase of phagocytosis.
单核细胞增生李斯特菌是一种兼性胞内病原体,能够在吞噬细胞内存活和生长。一些兼性胞内细菌已被证明通过产生一组应激蛋白来应对吞噬细胞内的恶劣环境。由于单核细胞增生李斯特菌具有与其他细菌不同的胞内存活机制,我们研究了该细菌对巨噬细胞吞噬作用的表型反应。J774-1巨噬细胞吞噬单核细胞增生李斯特菌EGD后,在5小时的孵育期内,该微生物数量迅速增加了约20倍。在吞噬作用的这个阶段,通过二维凝胶电泳观察到32种蛋白质的选择性诱导。还研究了对热和过氧化氢环境应激的反应,发现诱导了14种热休克蛋白和13种氧化应激蛋白。其中5种诱导蛋白在热应激和氧化应激中都有。通过氨基酸测序分析,在热休克蛋白中证实了DnaK和GroEL的同源物。二维凝胶放射自显影片的比较显示,这些应激蛋白中没有一种是巨噬细胞内单核细胞增生李斯特菌诱导的蛋白质。这种行为与其他兼性胞内病原体的行为完全不同。在本研究中,细胞内生长的单核细胞增生李斯特菌中不存在已知由环境应激诱导的应激蛋白。这种缺失可能是由于微生物在吞噬作用的早期阶段迅速逃离这种应激环境的机制。