1 Department of Medical Microbiology and Immunology, Faculty of Health Sciences, Aarhus University, Aarhus University2 unknown3 Department of Biomedicine - Forskning og uddannelse, Øst, Department of Biomedicine, Health, Aarhus University4 Department of Biomedicine - Forskning og uddannelse, Øst, Department of Biomedicine, Health, Aarhus University
PURPOSE. To investigate the integrity of secretory IgA (S-IgA) in tear fluid during bacterial keratitis and to evaluate the significance of specific Pseudomonas aeruginosa extracellular proteases in the observed degradation of S-IgA. METHODS. The integrity of component chains of S-IgA in tear fluid from patients with keratitis caused by P. aeruginosa, Streptococcus group G, Moraxella catarrhalis, Staphylococcus aureus, coagulase-negative staphylococci, and the IgA1 protease-producing Streptococcus pneumoniae were compared with S-IgA in tear fluid, colostrum, and saliva from healthy individuals, and with tear S-IgA incubated with clinical isolates and genetically engineered P. aeruginosa strains with different protease profiles. Degradation of S-IgA and the significance of its glycosylation were analyzed in Western blots developed with antibodies against individual chains of S-IgA. RESULTS. Secretory component (SC) of S-IgA secreted by the lacrimal glands was more abundantly glycosylated than SC in colostrum and saliva. S-IgA degradation was only observed in tears from P. aeruginosa-infected patients and in a pattern similar to that observed by incubation of tear samples with P. aeruginosa strains and an extent closely correlated with their expression and activity of individual proteases. Experiments using isogenic mutants of P. aeruginosa PAO1 lacking either elastase or alkaline protease indicate that several proteases work in concert. CONCLUSIONS. Surprisingly, SC of tear S-IgA is more abundantly glycosylated than SC of S-IgA in other secretions, a difference of potentially great functional significance. Primarily SC and alpha-chains are partially degraded in vivo during pseudomonas keratitis by the concerted action of several proteases including elastase and alkaline protease.
Investigative Ophthalmology and Visual Science, 2008, Vol 49, p. 4939-44