Adherence of Pseudomonas aeruginosa to respiratory epithelium and the effect of leucocyte elastase.

The tracheobronchial secretions from patients with cystic fibrosis often contain high amounts of free proteases. To evaluate whether human leucocyte elastase (HLE) can favour the persistence of bacterial airways infection, we exposed the frog palate mucosa to HLE and then to radiolabelled Pseudomonas aeruginosa and followed the sequence of events by scanning electronmicroscopy. In response to HLE there was a marked outpouring of mucus and a desquamation of the epithelium. P. aeruginosa was shown to adhere to recently secreted granules of mucus and to the exposed submucosal underlying connective tissues. For the eight different bacterial strains studied, a significative adherence to HLE-injured mucosa was observed only in strains that possessed internal haemagglutinating activity. Neither the presence of fimbriae, nor of the mucoid exopolysaccharide, nor of the bacterial surface haemagglutinating activity could be related to adherence of P. aeruginosa to the injured mucosa. These results support the hypothesis that HLE enhances bacterial infection of the respiratory mucosa both by inducing mucus hypersecretion and by exposing receptors to the microbial adhesins. It is also suggested that P. aeruginosa internal lectins may be implicated in adherence to host tissues.

The frog palate mucosa as a model for studying bacterial adhesion to mucus-coated respiratory epithelium.

Most of the methods proposed to quantify bacterial adherence to respiratory mucosa differ mainly from in vivo conditions in the absence of the mucus blanket and in the exposure of the sub-mucosal connective tissue (SMCT) to the micro-organisms. We propose the frog palate as a model to study bacterial adhesion to the respiratory mucosa, with a system which allows the mucus to be preserved and the bacterial adhesion to be quantified in a standardized mucosal area, where mucociliary transport is still active. In order to evaluate the role of respiratory mucus in bacteria-mucosa interaction, we compared the adhesion of radiolabelled pneumococci to 12 mucus-coated and 10 non-mucus-coated frog palate mucosae. The presence or absence of mucus was controlled by scanning electron microscopy (SEM). After a 10 min incubation period, the bacterial adhesion to mucus-coated palate mucosa was five times greater (P less than 0.01) than that to uncoated mucosa. By SEM, bacteria were never seen attached to ciliated cells but could be detected on small areas where mucus was not totally eliminated. Even after a 120 min contact of bacteria to uncoated mucosa, bacterial adhesion remained only half that to mucus-coated epithelium. In order to ascertain whether the exposure of the SMCT represented a means of attraction to bacteria, we incubated the frog palate mucosa face-down with radiolabelled Pseudomonas aeruginosa. As much as 44 per cent of added bacteria adhered to exposed SMCT and, by SEM, numerous micro-organisms were seen attached to connective tissue. In contrast, only a few bacteria were observed adhering to the mucosa, mainly to granules of mucus.

A new model for studying bacterial adherence to the respiratory epithelium.

The frog palate mucosa was used as a new model for studying bacterial adherence to the respiratory epithelium. The main advantage of this model is that the mucus blanket, normally present on airway mucosa, can be preserved during the assays. The adherence of radiolabeled pneumococci to mucus-coated mucosa was five times higher (P less than 0.001) than the adherence to mucus-depleted mucosa. In the latter case, bacteria were never seen attached to ciliated cells but could be detected on small remaining patches of mucus. These results demonstrate that respiratory mucus plays a major role in bacteria-mucosa interactions.

Evaluation of the 99mtechnetium labelling effect on Pseudomonas aeruginosa surface properties.

Gamma emitter isotopes present some advantages over beta emitters as radioisotopic microbial labels. The labelling of bacteria with 99mtechnetium (99mTc) has recently been described. However, it was not ascertained whether the labelling process modifies microbial physicochemical surface properties important in the interaction between bacteria and eukaryotic cells. In the present study, we evaluated the effect of the labelling process on Pseudomonas aeruginosa surface charge, hydrophobicity, adherence to human buccal epithelial cells and phagocytosis by human leukocytes. No significant differences in electrophoretic mobility or cationized ferritin distribution was observed on the cell surface of labelled and unlabelled bacteria. 99mTc labelling did not modify the hydrophobicity adhesiveness or phagocytosis of P. aeruginosa. It is concluded that bacterial labelling with 99mTc may be a useful method for the numeration of bacteria and the analysis of their functional properties.

The 99m technetium labeling effect on bacterial surface properties.

The present study was undertaken to determine whether labeling Pseudomonas aeruginosa with 99m Technetium would modify some of the bacterial physicochemical surface properties which play an important role in the interaction between bacteria and eukaryotic cells. No significant difference in electrophoretic mobility or distribution of cationized ferritin on the cell surface was observed between labeled and unlabeled bacteria. Also, the 99m Tc labeling process did not modify bacterial hydrophobicity or adhesiveness to human buccal epithelial cells. It is concluded that bacterial labeling with 99m Tc can be accepted as a useful method for biological research.

The relationship of the rate of intrauterine growth of low-birth-weight infants to later growth.

The relationship of the rate of intrauterine growth of low-birth-weight infants (1,501 to 2,500 gm) to their postnatal growth up to 10 years of age was investigated. Each child was assigned to one of four gestation quartiles which have identical birth weight distributions but differ widely in their length of gestation. The mean heights and weights of the children in each of the four quartiles were compared with similar data of a control group of children who had birth weights above 2,500 gm. Only infants in the long gestation quartile failed to catch up in height and weight to the control group by the end of the 10-year period. The same results that were obtained for the total cohort were also found separately for white and black children and for boys and girls. The data relate to 488 single white and black low-birth-weight infants and 488 normal-birth-weight infants.