Scharfman A, Kroczynski H, Carnoy C, Van Brussel E, Lamblin G, Ramphal R, Roussel P
Unité INSERM 377, Lille, France.
Infect Immun. 1996 Dec;64(12):5417-20. doi: 10.1128/iai.64.12.5417-5420.1996.
The attachment of Pseudomonas aeruginosa to human respiratory mucus represents an important step in the development of lung infection, especially in cystic fibrosis. Local factors in the respiratory tract, such as osmolarity or iron concentration, might influence this colonization. In the present work, we have observed that overall levels of adhesion of two nonmucoid, nonpiliated strains of P. aeruginosa, 1244-NP and PAK-NP, to human airway mucins were higher when these strains were grown in a minimal medium of low osmolarity (M9) than when they were grown in a rich medium of higher osmolarity (tryptic soy broth [TSB]). However, increasing the NaCl concentration of M9 to increase the osmolarity did not modify the level of binding. In order to find out whether these differences were due to variations in nutrients, the influence of iron concentration was investigated: the levels of binding of the two strains increased after TSB was depleted of iron and decreased after iron was added to M9. Since the outer membranes from the two strains have been shown to contain proteins reacting with human bronchial mucins, we compared the mucin-binding proteins expressed by the two strains grown in different media. When the nonpiliated strains 1244-NP and PAK-NP were grown in the different media, previously observed mucin-binding bands were detected in the 42- to 48-kDa range but additional mucin-binding bands in the 77- to 85-kDa range were detected when these strains were grown in M9 or iron-deprived TSB. These results demonstrate that the adhesion of P. aeruginosa and the expression of mucin-binding proteins in the outer membranes of nonpiliated P. aeruginosa are affected by the iron content of the medium in which the bacteria are grown and not by the osmolarity.
铜绿假单胞菌附着于人类呼吸道黏液是肺部感染发展过程中的重要一步,尤其是在囊性纤维化患者中。呼吸道中的局部因素,如渗透压或铁浓度,可能会影响这种定植。在本研究中,我们观察到,两种非黏液型、无纤毛的铜绿假单胞菌菌株1244-NP和PAK-NP,当它们在低渗透压的基本培养基(M9)中生长时,与人类气道黏蛋白的总体黏附水平高于在高渗透压的丰富培养基(胰蛋白胨大豆肉汤[TSB])中生长时。然而,增加M9中的NaCl浓度以提高渗透压并没有改变结合水平。为了确定这些差异是否是由于营养成分的变化引起的,我们研究了铁浓度的影响:TSB中铁耗尽后,这两种菌株的结合水平增加,而向M9中添加铁后,结合水平降低。由于已证明这两种菌株的外膜含有与人类支气管黏蛋白反应的蛋白质,我们比较了在不同培养基中生长的这两种菌株所表达的黏蛋白结合蛋白。当无纤毛菌株1244-NP和PAK-NP在不同培养基中生长时,在42至48 kDa范围内检测到先前观察到的黏蛋白结合条带,但当这些菌株在M9或缺铁的TSB中生长时,在77至85 kDa范围内检测到额外的黏蛋白结合条带。这些结果表明,铜绿假单胞菌的黏附以及无纤毛铜绿假单胞菌外膜中黏蛋白结合蛋白的表达受细菌生长培养基中铁含量的影响,而不受渗透压的影响。