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SIgA binding to mucosal surfaces is mediated by mucin-mucin interactions.

Gibbins HL, Proctor GB, Yakubov GE, Wilson S, Carpenter GH - PLoS ONE (2015)

Bottom Line: Binding of SIgA to cells was then compared using whole mouth saliva, parotid (mucin-free) saliva and a source of purified SIgA.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.

View Article: PubMed Central - PubMed

Affiliation: Salivary Research Unit, King's College London Dental Institute, London, United Kingdom.

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.

No MeSH data available.


Concentration of IgA bound to equal protein levels of HT29 and HT29-MTX cell lines from UWMS at days 4, 8 and 16.Significantly more IgA is bound to HT29-MTX cells compared to HT29 cells at day 16 and also significantly more compared to the same cell line at day 4, where *P<0.05 Day 16 HT29-MTX compared to day 4 HT29-MTX, **P<0.01 day 16 HT29-MTX compared to Day 16 HT29.
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pone.0119677.g003: Concentration of IgA bound to equal protein levels of HT29 and HT29-MTX cell lines from UWMS at days 4, 8 and 16.Significantly more IgA is bound to HT29-MTX cells compared to HT29 cells at day 16 and also significantly more compared to the same cell line at day 4, where *P<0.05 Day 16 HT29-MTX compared to day 4 HT29-MTX, **P<0.01 day 16 HT29-MTX compared to Day 16 HT29.

Mentions: Following a 1 hour incubation of UWMS with both cell lines the amount of IgA bound to the cells was much greater in the presence of the MUC5AC on the HT29-MTX cells at day 8 and day 16, as seen in Fig. 3. At day 16 IgA bound to the HT29-MTX cells was over 8x the amount bound to the HT29 cells (p<0.01). This was significantly more than bound to the HT29-MTX cells at day 4 where there was no MUC5AC mucus development (p<0.05). With a 20 minute incubation with UWMS, at both day 8 and day 16 there was significantly more IgA binding onto the HT29-MTX cells (P<0.05 and P<0.01), in comparison to the HT29 cells without the mucus layer (data not shown).


SIgA binding to mucosal surfaces is mediated by mucin-mucin interactions.

Gibbins HL, Proctor GB, Yakubov GE, Wilson S, Carpenter GH - PLoS ONE (2015)

Concentration of IgA bound to equal protein levels of HT29 and HT29-MTX cell lines from UWMS at days 4, 8 and 16.Significantly more IgA is bound to HT29-MTX cells compared to HT29 cells at day 16 and also significantly more compared to the same cell line at day 4, where *P<0.05 Day 16 HT29-MTX compared to day 4 HT29-MTX, **P<0.01 day 16 HT29-MTX compared to Day 16 HT29.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4368717&req=5

pone.0119677.g003: Concentration of IgA bound to equal protein levels of HT29 and HT29-MTX cell lines from UWMS at days 4, 8 and 16.Significantly more IgA is bound to HT29-MTX cells compared to HT29 cells at day 16 and also significantly more compared to the same cell line at day 4, where *P<0.05 Day 16 HT29-MTX compared to day 4 HT29-MTX, **P<0.01 day 16 HT29-MTX compared to Day 16 HT29.
Mentions: Following a 1 hour incubation of UWMS with both cell lines the amount of IgA bound to the cells was much greater in the presence of the MUC5AC on the HT29-MTX cells at day 8 and day 16, as seen in Fig. 3. At day 16 IgA bound to the HT29-MTX cells was over 8x the amount bound to the HT29 cells (p<0.01). This was significantly more than bound to the HT29-MTX cells at day 4 where there was no MUC5AC mucus development (p<0.05). With a 20 minute incubation with UWMS, at both day 8 and day 16 there was significantly more IgA binding onto the HT29-MTX cells (P<0.05 and P<0.01), in comparison to the HT29 cells without the mucus layer (data not shown).

Bottom Line: Binding of SIgA to cells was then compared using whole mouth saliva, parotid (mucin-free) saliva and a source of purified SIgA.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.

View Article: PubMed Central - PubMed

Affiliation: Salivary Research Unit, King's College London Dental Institute, London, United Kingdom.

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.

No MeSH data available.