Hauf N, Goebel W, Serfling E, Kuhn M
Lehrstuhl für Mikrobiologie, Theodor-Boveri-Institut für Biowissenschaften, Universität Würzburg, Germany.
Infect Immun. 1994 Jul;62(7):2740-7. doi: 10.1128/iai.62.7.2740-2747.1994.
In the present study, we investigated the effect of Listeria monocytogenes infection on the cellular level of the transcription factors NF-kappa B, AP-1, and NF-IL6 in the macrophage-like cell line P388D1 by using electrophoretic mobility shift assays. Infection with L. monocytogenes enhanced the formation of two NF-kappa B-like DNA-protein complexes, C1 and C2, whereas the concentration of AP-1 and NF-IL6 complexes remained unaffected. In supershift assays using NF-kappa B-specific antibodies, complex C2 was identified to be a p50 homodimer (KBF1) and complex C1 was identified as a p50/p65 heterodimer. Both complexes were formed within 10 min after addition of the bacteria. Since the synthesis of tumor necrosis factor alpha and interleukin-1 occurs at later times, these cytokines cannot be the mediators of enhanced NF-kappa B formation. Infection experiments with different nonhemolytic mutants of L. monocytogenes and the use of the phagocytosis inhibitor cytochalasin B suggest that events prior to invasion and escape of the bacteria from the phagosome into the cytoplasm enhance the nuclear transport of p50/p65 NF-kappa B components.
在本研究中，我们通过电泳迁移率变动分析，研究了单核细胞增生李斯特菌感染对巨噬细胞样细胞系P388D1中转录因子NF-κB、AP-1和NF-IL6细胞水平的影响。单核细胞增生李斯特菌感染增强了两种NF-κB样DNA-蛋白质复合物C1和C2的形成，而AP-1和NF-IL6复合物的浓度未受影响。在使用NF-κB特异性抗体的超迁移分析中，复合物C2被鉴定为p50同二聚体（KBF1），复合物C1被鉴定为p50/p65异二聚体。两种复合物在加入细菌后10分钟内形成。由于肿瘤坏死因子α和白细胞介素-1的合成在较晚时间发生，这些细胞因子不可能是NF-κB形成增强的介质。用单核细胞增生李斯特菌不同的非溶血性突变体进行的感染实验以及吞噬作用抑制剂细胞松弛素B的使用表明，在细菌从吞噬体侵入并逃逸到细胞质之前的事件增强了p50/p65 NF-κB组分的核转运。
