Detection of secretory IgM in tears of IgA deficient individuals.
Tears from normal (n = 5) and serum IgA deficient (n = 3) individuals were investigated for the presence of secretory Immunoglobulin A (sIgA), sIgM and free secretory component (SC) by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) using 10-15% gradient minigels (PhastSystem), followed by immunoblotting using various immunological probes. Tear samples were treated in denaturing (SDS) sample buffer under non-reducing as well as reducing conditions, prior to analysis. All normal tear samples contained sIgA as well as free SC (estimated MW: 82kD) but only traces of IgM. Tears from the three serum IgA deficient subjects lacked sIgA but did contain free SC. In two of them sIgM was clearly detected and after treatment of tears with reducing agent, IgM (mu) heavy chain fragments (estimated MW: 78kD) were identified and could be distinguished from other tear proteins after SDS-PAGE. These findings indicate lacrimal secretion of free secretory component, even in the absence of its ligand. On the ocular surface, sIgM may play a compensatory role in IgA deficiency.
Kuizenga
A
A
Biochemical Laboratory, The Netherlands Ophthalmic Research Institute, Amsterdam.
Stolwijk
T R
TR
van Agtmaal
E J
EJ
van Haeringen
N J
NJ
Kijlstra
A
A
D000328:Adult; D004406:Dysgammaglobulinemia / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D017098:IgA Deficiency; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007070:Immunoglobulin A / Q000276:immunology; D007071:Immunoglobulin A, Secretory / Q000032:analysis; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D007075:Immunoglobulin M / Q000032:analysis*; D008297:Male; D008970:Molecular Weight; D012635:Secretory Component / Q000032:analysis; D013666:Tears / Q000276:immunology*
D007070:Immunoglobulin A; D007071:Immunoglobulin A, Secretory; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M; D012635:Secretory Component; C033027:secretory IgM