Brock J H, Williams P H, Licéaga J, Wooldridge K G
University Department of Bacteriology and Immunology, Western Infirmary, Glasgow, United Kingdom.
Infect Immun. 1991 Sep;59(9):3185-90. doi: 10.1128/iai.59.9.3185-3190.1991.
A method is described for determination of the relative availability of transferrin-bound iron and cell-derived iron to microbial iron-scavenging mechanisms. This involved incubation of parallel cultures of microorganisms in dialysis tubes placed in RPMI 1640 tissue culture medium containing 30%-iron-saturated transferrin and K562 erythroleukemia cells. In one culture the transferrin was labelled with 59Fe and in the other the cells were labelled, and the relative uptake of radioiron by the microorganisms determined. The results showed that Staphylococcus epidermidis and Staphylococcus aureus acquired iron predominantly from cells, while Candida albicans and the enteropathogenic Escherichia coli NCTC 8623 tended to acquire iron from transferrin. E. coli K-12 strains W3110 and LG1705, which (like NCTC 8623) produce the siderophore enterochelin but not aerobactin, acquired predominantly transferrin-bound iron, whereas the related E. coli strains LG1315 and LG1628, which produce aerobactin but not enterochelin, showed a preference for cell-derived iron. When the cells were incubated in the presence of 59Fe-labelled transferrin and 55Fe-labelled ferritin, no difference in relative availability of iron to E. coli was observed, suggesting that differences in the ability of aerobactin and enterochelin to remove iron from intracellular ferritin were not responsible for this preference. These results may help to explain why production of aerobactin, despite its relatively low affinity for iron, is more closely associated with invasiveness in E. coli than is enterochelin production. Reduced availability of cell-bound iron during inflammation may contribute to antimicrobial defenses.
本文描述了一种测定转铁蛋白结合铁和细胞源性铁对微生物铁清除机制的相对可用性的方法。该方法包括将微生物的平行培养物置于含有30%铁饱和转铁蛋白和K562红白血病细胞的RPMI 1640组织培养基中的透析管中进行孵育。在一种培养物中转铁蛋白用59Fe标记,在另一种培养物中细胞被标记,然后测定微生物对放射性铁的相对摄取量。结果表明,表皮葡萄球菌和金黄色葡萄球菌主要从细胞中获取铁,而白色念珠菌和肠道致病性大肠杆菌NCTC 8623倾向于从转铁蛋白中获取铁。大肠杆菌K-12菌株W3110和LG1705(与NCTC 8623一样)产生铁载体肠螯合素但不产生气杆菌素,它们主要获取转铁蛋白结合的铁,而相关的大肠杆菌菌株LG1315和LG1628产生气杆菌素但不产生肠螯合素,它们更倾向于细胞源性铁。当细胞在59Fe标记的转铁蛋白和55Fe标记的铁蛋白存在下孵育时,未观察到大肠杆菌对铁的相对可用性有差异,这表明气杆菌素和肠螯合素从细胞内铁蛋白中去除铁的能力差异不是造成这种偏好的原因。这些结果可能有助于解释为什么尽管气杆菌素对铁的亲和力相对较低,但在大肠杆菌中它比肠螯合素的产生与侵袭性的关联更紧密。炎症期间细胞结合铁的可用性降低可能有助于抗菌防御。