Ethanolamine utilization supports Clostridium perfringens growth in infected tissues
Ethanolamine utilization supports Clostridium perfringens growth in infected tissues
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乙醇胺的利用支持产气荚膜梭菌在感染组织中的生长
DOI：
10.1016/j.micpath.2018.04.017
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发表时间：
2018
影响因子：
3.8
通讯作者：
Kuwahara Tomomi
中科院分区：
文献类型：
--
作者：
Yagi Hirofumi;Nakayama-Imaohji Haruyuki;Nariya Hirofumi;Tada Ayano;Yamasaki Hisashi;Ugai Hideyo;Elahi Miad;Ono Tsuneko;Kuwahara Tomomi
产气荚膜梭菌具有编码在乙醇胺（EA）利用（eut）操纵子内的eut系统，其利用EA作为碳、氮和能量来源。为了确定eut系统在C.为了抑制产气荚膜杆菌生长，在菌株HN 13中进行eutABC基因的框内缺失以产生eutABC缺失的突变菌株HY 1701。在添加1.0%葡萄糖和/或1.0%EA的培养基中比较HN 13和HY 1701的生长，发现葡萄糖促进了两株菌的生长，而EA促进了HN 13的生长，但对HY 1701的生长没有促进作用。产气荚膜杆菌利用EA。EutVW的两个组件的监管系统是需要诱导eut基因的表达响应EA，而全球毒力调节VirRS差异控制eut基因的表达取决于葡萄糖和EA的可用性。为了评估eut系统在体内的作用，将相同数量的HN 13和HY 1701细胞注射到小鼠的右大腿肌肉中。感染HY 1701的小鼠比注射HN 13的小鼠表现出更少的症状。感染HY 1701的小鼠的死亡率倾向于低于感染HN 13的小鼠。此外，在用HN 13和HY 1701的混合物注射的小鼠的感染组织中，HN 13的数量超过HY 1701。PCR筛选结果表明，C.从气性坏疽和散发性腹泻病例中分离的产气荚膜杆菌携带eut基因和产气荚膜溶素O基因（pfoA）以及磷脂酶C基因（plc）。然而，pfoA在食物中毒患者和健康志愿者的分离物中未检测到。与plc和/或pfoA缺失突变体相比，在含有7.5%绵羊红细胞的培养基中生长的HN 13制备的培养上清液诱导显著更高的eutB表达水平。总之，这些结果表明eut系统对C.溶组织感染期间产气荚膜杆菌。
Clostridium perfringens possesses the ethanolamine (EA) utilization (eut) system encoded within the eut operon, which utilizes the EA as a carbon, nitrogen and energy source. To determine the role of the eut system in C. perfringens growth, an in-frame deletion of the eutABC genes was made in strain HN13 to generate the eutABC-deleted mutant strain HY1701. Comparison of HN13 and HY1701 growth in media supplemented with 1.0% glucose and/or 1.0% EA showed that glucose enhanced the growth of both strains, whereas EA enhanced HN13 growth, but not that of HY1701, indicating that the eut system is necessary for C. perfringens to utilize EA. The two-component regulatory system EutVW is needed to induce eut gene expression in response to EA whereas the global virulence regulator VirRS differentially controlled eut gene expression depending on glucose and EA availability. To assess the role of the eut system in vivo, an equal number of HN13 and HY1701 cells were injected into the right thigh muscles of mice. Mice infected with HY1701 showed fewer symptoms than those injected with HN13. The mortality rate of mice infected with HY1701 tended to be lower than for mice infected with HN13. In addition, in infected tissues from mice injected with a mixture of HN13 and HY1701, HN13 outnumbered HY1701. PCR screening demonstrated that C. perfringens isolated from gas gangrene and sporadic diarrhea cases carried both eut genes and the perfringolysin O gene (pfoA) as well as the phospholipase C gene (plc). However, pfoA was not detected in isolates from food poisoning patients and healthy volunteers. Culture supernatants prepared from HN13 grown in media containing 7.5% sheep red blood cells induced significantly higher eutB expression levels compared to those from plc-and/or pfoA-deletion mutants. Together, these results indicate that the eut system plays a nutritional role for C. perfringens during histolytic infection.