Morphological and functional glycobiological analysis of human airway cilia in health and disease

Adam, Elizabeth Caroline (1998) Morphological and functional glycobiological analysis of human airway cilia in health and disease. University of Southampton, Doctoral Thesis.

Abstract

This thesis describes the development of a dedicated system for measuring and recording the frequency with which cilia beat together with analysis of the data, to provide a quantitative system for evaluation of modifications to ciliary beating. Human nasal polyps were used to provide beating cilia representative of the respiratory tract and a means of maintaining explants of this tissue in vitro for up to a month was developed.

Carbohydrate structures on the surface of the cilia were defined using lectins which are highly-specific carbohydrate-binding glycoproteins. As a result of these investigations several lectins were selected which bound to the surface of the ciliated cells. Using the test system developed, explants in culture were exposed to these lectins and one lectin in particular (Wheat Germ Agglutinin) caused a complete cessation of ciliary beating within 24 hours. This inhibition of ciliary activity was prevented by preincubation of the lectin with its inhibitory sugar N-acetylglucosamine.

This technique was extended to include bacterial lectins from the bacterium Pseudomonas aeruginosa which inhabits compromised respiratory tracts and is resistant to antibiotics. Very low doses of one of these lectins (PA-II) also resulted in total inhibition of ciliary activity within a few hours, unless it had been preincubated with its inhibitory sugar fucose when no changes were observed. Even more importantly, with the addition of fucose to the culture medium, it proved possible to intercept the action of PA-II even after the cilia had started to slow down. This has therapeutic implications for those individuals such as cystic fibrosis patients who suffer from recurrent respiratory infections with P. aeruginosa.

The system to measure ciliary beat frequency has also been applied in clinical studies where it is extremely difficult to diagnose the presence of a genetically-inherited condition known as Primary Ciliary Dyskinesia (PCD). This condition manifests itself as a range of abnormalities in ciliary ultrastructure, many of which result in reduction or stoppage of ciliary beating.

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