Du Yidong, Rosqvist Roland, Forsberg Ake
Division of NBC-Protection, Swedish Defence Research Agency, SE-901 82 Umeå, Sweden.
Infect Immun. 2002 Mar;70(3):1453-60. doi: 10.1128/IAI.70.3.1453-1460.2002.
Yersinia pestis, the causative agent of plague, expresses a capsule-like antigen, fraction 1 (F1), at 37 degrees C. F1 is encoded by the caf1 gene located on the large 100-kb pFra plasmid, which is unique to Y. pestis. F1 is a surface polymer composed of a protein subunit, Caf1, with a molecular mass of 15.5 kDa. The secretion and assembly of F1 require the caf1M and caf1A genes, which are homologous to the chaperone and usher protein families required for biogenesis of pili. F1 has been implicated to be involved in the ability of Y. pestis to prevent uptake by macrophages. In this study we addressed the role of F1 antigen in inhibition of phagocytosis by the macrophage-like cell line J774. The Y. pestis strain EV76 was found to be highly resistant to uptake by J774 cells. An in-frame deletion of the caf1M gene of the Y. pestis strain EV76 was constructed and found to be unable to express F1 polymer on the bacterial surface. This strain had a somewhat lowered ability to prevent uptake by J774 cells. Strain EV76C, which is cured for the virulence plasmid common to the pathogenic Yersinia species, was, as expected, much reduced in its ability to resist uptake. A strain lacking both the virulence plasmid and caf1M was even further hampered in the ability to prevent uptake and, in this case, essentially all bacteria (95%) were phagocytosed. Thus, F1 and the virulence plasmid-encoded type III system act in concert to make Y. pestis highly resistant to uptake by phagocytes. In contrast to the type III effector proteins YopE and YopH, F1 did not have any influence on the general phagocytic ability of J774 cells. Expression of F1 also reduced the number of bacteria that interacted with the macrophages. This suggests that F1 prevents uptake by interfering at the level of receptor interaction in the phagocytosis process.
鼠疫耶尔森菌是鼠疫的病原体,在37摄氏度时表达一种类荚膜抗原,即1号组分(F1)。F1由位于100 kb的大质粒pFra上的caf1基因编码,该质粒是鼠疫耶尔森菌所特有的。F1是一种表面聚合物,由分子量为15.5 kDa的蛋白质亚基Caf1组成。F1的分泌和组装需要caf1M和caf1A基因,它们与菌毛生物合成所需的伴侣蛋白家族和外膜蛋白家族同源。F1被认为与鼠疫耶尔森菌阻止被巨噬细胞摄取的能力有关。在本研究中,我们探讨了F1抗原在抑制巨噬细胞样细胞系J774吞噬作用中的作用。发现鼠疫耶尔森菌菌株EV76对J774细胞的摄取具有高度抗性。构建了鼠疫耶尔森菌菌株EV76的caf1M基因的框内缺失突变体,发现其无法在细菌表面表达F1聚合物。该菌株阻止被J774细胞摄取的能力有所降低。菌株EV76C缺失了致病性耶尔森菌属共有的毒力质粒,正如预期的那样,其抵抗摄取的能力大大降低。一个既缺乏毒力质粒又缺乏caf1M的菌株在阻止摄取的能力上受到了更大的阻碍,在这种情况下,基本上所有细菌(95%)都被吞噬了。因此,F1和毒力质粒编码的III型分泌系统协同作用,使鼠疫耶尔森菌对吞噬细胞的摄取具有高度抗性。与III型效应蛋白YopE和YopH不同,F1对J774细胞的一般吞噬能力没有任何影响。F1的表达也减少了与巨噬细胞相互作用的细菌数量。这表明F1通过在吞噬过程中干扰受体相互作用水平来阻止摄取。