Abstract

The oral mucosal pellicle is a layer of absorbed salivary proteins, including secretory IgA (SIgA), bound onto the surface of oral epithelial cells and is a useful model for all mucosal surfaces. The mechanism by which SIgA concentrates on mucosal surfaces is examined here using a tissue culture model with real saliva. Salivary mucins may initiate the formation of the mucosal pellicle through interactions with membrane-bound mucins on cells. Further protein interactions with mucins may then trigger binding of other pellicle proteins. HT29 colon cell lines, which when treated with methotrexate (HT29-MTX) produce a gel-forming mucin, were used to determine the importance of these mucin-mucin interactions. Binding of SIgA to cells was then compared using whole mouth saliva, parotid (mucin-free) saliva and a source of purified SIgA. Greatest SIgA binding occurred when WMS was incubated with HT29-MTX expressing mucus. Since salivary MUC5B was only able to bind to cells which produced mucus and purified SIgA showed little binding to the same cells we conclude that most SIgA binding to mucosal cells occurs because SIgA forms complexes with salivary mucins which then bind to cells expressing membrane-bound mucins. This work highlights the importance of mucin interactions in the development of the mucosal pellicle.

Highlights

  • The mucus layer is essential for protection, molecular transport and lubrication on soft tissues and linings of most of the essential organs

  • [25] MUC5B was demonstrated to be strongly retained on buccal cells thereby showing its crucial role in pellicle formation on soft oral tissues

  • It is known that HT29 cells have suppressed expression of membrane bound MUC1 mucins [27], which otherwise covers most mammalian cells including the oral buccal cells that were tested as being attractive for MUC5B deposition [28]

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Summary

Introduction

The mucus layer is essential for protection, molecular transport and lubrication on soft tissues and linings of most of the essential organs. In airways and gastrointestinal tract the mucosal film is formed primarily by mucins, while in other linings like that in the oral cavity the mucosal film (salivary pellicle) contains globular proteins and proline-rich proteins. Among these globular proteins secretory IgA (SIgA) plays an important role in topical immune response of the adsorbed proteinaceous film. While mucins spontaneously assemble on mucosal surfaces in vivo, this behaviour has never yet been fully replicated in vitro using purified mucins. The inability to replicate the mucosal layer stems from two key factors. Purification of proteins leads to the loss of their tertiary conformation, even if mucin preparations

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