Normal but not altered mucins activate neutrophils.
Interactions between leucocytes and their surroundings are mediated through oligosaccharide epitopes, some of which are also expressed on ocular mucins. Neutrophils represent the majority of immune cells in the proinflammatory environment of the ocular surface during sleep. We have tested whether changes in mucin glycosylation, as occur in dry eyes, influence the phenotype and activation of neutrophils. Peripheral blood leucocytes were circulated over equal concentration mats of ocular surface mucins purified from normal volunteers and dry-eye patients, and in sequence over normal and pathological mucins in all combinations. Non-adherent cells were tagged with monoclonal fluorescent antibodies to leucocyte determinants and analysed by flow cytometry. Oxidative burst, assessed with dihydrorhodamine, was followed in cells and supernatant. At a speed similar to that of leucocyte traffic in the retina, normal mucins caused a decrease in neutrophil cathepsin G fluorescence, a decrease that was not observed with mucins from patients with Meibomian gland disease or Sjogren syndrome. No effect was detected at a higher flow. Supernatant and cells collected after circulation over normal mucin showed increased rhodamine fluorescence, indicative of oxidative burst. Fluorescence could also be observed in intact cells adherent to dry-eye mucins. Non-adherent cells could be activated with phorbol 12-myristate 13-acetate after flow over any mucin or combination of mucins. Differences in neutrophil activation after exposure to normal and pathological mucins highlight reciprocal influences at the interface between local and systemic immunity.
Aknin
Marie-Laure Rachel
ML
Department of Clinical Sciences, Ophthalmology, Bristol Eye Hospital, University of Bristol, BS1 2LX, Bristol, UK. ml.aknin@bristol.ac.uk
Berry
Monica
M
Dick
Andrew David
AD
Khan-Lim
Doreen
D
D000328:Adult; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D005141:Eyelid Diseases / Q000097:blood; D005434:Flow Cytometry; D006031:Glycosylation; D006801:Humans; D008537:Meibomian Glands / Q000378:metabolism / Q000473:pathology; D008875:Middle Aged; D009077:Mucins / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D018375:Neutrophil Activation / Q000187:drug effects* / Q000276:immunology; D009504:Neutrophils / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D016897:Respiratory Burst / Q000187:drug effects / Q000276:immunology; D012859:Sjogren's Syndrome / Q000097:blood
D009077:Mucins