Disease and Carrier Isolates of Neisseria meningitidis Cause G1 Cell Cycle Arrest in Human Epithelial Cells
Disease and Carrier Isolates of Neisseria meningitidis Cause G1 Cell Cycle Arrest in Human Epithelial Cells
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DOI：
10.1128/iai.00296-16
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发表时间：
2016-07
影响因子：
3.1
通讯作者：
Michael von Papen;W. Oosthuysen;J. Bécam;H. Claus;A. Schubert-Unkmeir
中科院分区：
文献类型：
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作者：
Michael von Papen;W. Oosthuysen;J. Bécam;H. Claus;A. Schubert-Unkmeir
摘要微生物病原体已经发展出几种机制来调节和干扰宿主细胞周期进程。在这项研究中，我们分析了人类病原体脑膜炎奈瑟菌对上皮细胞周期的影响。对两个致病菌株和两个携带菌株进行了黏附和侵袭底特律562和NP69上皮细胞的能力以及调节细胞周期的能力的测试。我们发现，所有的菌株对底特律562和NP69细胞的粘附性都一样好，而携带者的菌株侵袭性要小得多。用碘化丙啶染色和5-乙炔基-2‘-脱氧尿嘧啶核苷脉冲标记，我们提供了脑膜炎球菌感染使细胞在感染后24小时停滞在细胞周期的G1期的证据。同时，观察到S期细胞明显减少。有趣的是，只有在感染活细菌后才会诱导G1期停滞，而热致死细菌则不会。通过Western blotting，我们发现细菌感染导致细胞周期调节因子细胞周期蛋白D1的蛋白水平降低，而细胞周期蛋白E的表达水平增加。此外，免疫荧光分析表明，脑膜炎奈瑟氏菌感染诱导了细胞周期蛋白依赖性激酶抑制因子(CKI)p21WAF1/CIP1的积聚，并伴随着该CKI重新分布到细胞核。此外，p27CIP1 CKI在感染细胞中重新分布，呈点状分布。综上所述，我们提供了脑膜炎奈瑟氏菌干扰宿主细胞周期调节过程的数据。
ABSTRACT Microbial pathogens have developed several mechanisms to modulate and interfere with host cell cycle progression. In this study, we analyzed the effect of the human pathogen Neisseria meningitidis on the cell cycle of epithelial cells. Two pathogenic isolates, as well as two carrier isolates, were tested for their ability to adhere to and invade into the epithelial cell lines Detroit 562 and NP69 and to modulate the cell cycle. We found that all isolates adhered equally well to both Detroit 562 and NP69 cells, whereas the carrier isolates were significantly less invasive. Using propidium iodide staining and 5-ethynyl-2′-deoxyuridine pulse-labeling, we provide evidence that meningococcal infection arrested cells in the G1 phase of the cell cycle at 24 h postinfection. In parallel, a significant decrease of cells in the S phase was observed. Interestingly, G1-phase arrest was only induced after infection with live bacteria but not with heat-killed bacteria. By Western blotting we demonstrate that bacterial infection resulted in a decreased protein level of the cell cycle regulator cyclin D1, whereas cyclin E expression levels were increased. Furthermore, N. meningitidis infection induced an accumulation of the cyclin-dependent kinase inhibitor (CKI) p21WAF1/CIP1 that was accompanied by a redistribution of this CKI to the cell nucleus, as shown by immunofluorescence analysis. Moreover, the p27CIP1 CKI was redistributed and showed punctate foci in infected cells. In summary, we present data that N. meningitidis can interfere with the processes of host cell cycle regulation.