Valentin-Weigand P, Benkel P, Rohde M, Chhatwal G S
Department of Microbiology, GBF-National Research Centre for Biotechnology, Braunschweig, Germany.
Infect Immun. 1996 Jul;64(7):2467-73. doi: 10.1128/iai.64.7.2467-2473.1996.
The mouse macrophage-like cell line J774 was used to analyze opsonin-independent entry and survival of group B streptococci (GBS). Efficient entry of GBS in J774 cells occurred within 5 min postinfection, and streptococci persisted intracellularly without loss of viability for at least 8 h. At 24 h postinfection, 30% of the total intracellular GBS was recovered from macrophages. Inhibition studies using different biochemical modulators of cellular functions showed that bacterial entry seemed to involve nonglycosylated J774 surface structures different from known receptors such as fibronectin-binding integrins. Internalization of GBS by J774 cells occurred by a microfilament-dependent phagocytosis-like process also involving participation of receptor-mediated endocytosis. Prior opsonization of GBS with human serum containing anti-GBS antibodies did not affect bacterial entry but significantly reduced the intracellular survival of GBS. Transmission electron microscopic analysis confirmed these findings and demonstrated that both opsonized and nonopsonized bacteria were contained within phagosomes during the whole infection period. Transmission electron microscopy further revealed that decreased intracellular survival rates of opsonized GBS appeared to be due to increased lysosomal activities of the macrophages. These results suggest that in the absence of opsonins, GBS are able to enter and persist efficiently in macrophages by evading intracellular antibacterial activities commonly associated with opsonin-mediated uptake.
小鼠巨噬细胞样细胞系J774用于分析B族链球菌(GBS)的非调理素依赖性进入和存活情况。GBS在感染后5分钟内即可高效进入J774细胞,并且链球菌在细胞内存活至少8小时,活力无损失。感染后24小时,从巨噬细胞中回收了30%的细胞内GBS。使用不同细胞功能生化调节剂的抑制研究表明,细菌进入似乎涉及与已知受体(如纤连蛋白结合整合素)不同的非糖基化J774表面结构。J774细胞对GBS的内化通过微丝依赖性吞噬样过程发生,该过程也涉及受体介导的内吞作用。用含抗GBS抗体的人血清预先调理GBS不影响细菌进入,但显著降低了GBS在细胞内的存活率。透射电子显微镜分析证实了这些发现,并表明在整个感染期间,调理和未调理的细菌都被包裹在吞噬体中。透射电子显微镜进一步显示,调理GBS细胞内存活率降低似乎是由于巨噬细胞溶酶体活性增加。这些结果表明,在缺乏调理素的情况下,GBS能够通过逃避通常与调理素介导的摄取相关的细胞内抗菌活性,有效地进入巨噬细胞并在其中存活。