High binding of immunoglobulin M kappa rheumatoid factor from type II cryoglobulins to cellular fibronectin: a mechanism for induction of in situ immune complex glomerulonephritis?
In our previous experimental work we suggested that the frequent nephritogenicity of type II cryoglobulins could depend on a particular affinity of the immunoglobulin (Ig) M kappa rheumatoid factor (RF) component for mesangial matrix. Since cellular fibronectin (cFN) in the human kidney is mainly represented in glomerular mesangium, we studied the binding capacity to cFN of IgM kappa RFs from type II cryoglobulins compared with other different monoclonal and polyclonal IgM and IgM RFs. We purified 13 IGM kappa from human IgM kappa/IgG cryoglobulins, eight monoclonal IgM from patients with Waldenström's macroglobulinemia, nine polyclonal IgM from normal donors, and eight polyclonal IgM RFs from patients with rheumatoid arthritis. Purified IgM were used at the same concentration in enzyme-linked immunosorbent assay (ELISA) on cFN-coated plates. All the cryoglobulin IgM showed high specific binding to cFN while IgM from Waldenström's macroglobulinemia, normal IgM, and polyclonal IgM RFs had low or absent binding. These data were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of cFN followed by Western blot analysis with purified IgM. The IgM kappa binding to cFN persisted using IgM kappa monomers, and was inhibited by cFN but not by plasma FN in a specific inhibition test. Further enzyme-linked immunosorbent assay studies showed that cryoglobulin IgM kappa RFs are still able to bind IgG in a dose-dependent manner once linked to solid-phase cFN. The data suggest that the affinity of cryoglobulin IgM kappa RFs for immobilized cFN could be involved in the particular high nephritogenicity of type II cryoglobulins and might lead to in situ immune complex formation.
Fornasieri
A
A
Division of Nephrology, San Carlo Hospital, Milano, Italy.
Armelloni
S
S
Bernasconi
P
P
Li
M
M
de Septis
C P
CP
Sinico
R A
RA
D'Amico
G
G
D001323:Autoantibodies / Q000378:metabolism; D001666:Binding Sites, Antibody / Q000502:physiology; D015153:Blotting, Western / Q000379:methods; D003449:Cryoglobulinemia / Q000378:metabolism; D003450:Cryoglobulins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005921:Glomerulonephritis / Q000209:etiology / Q000378:metabolism*; D006801:Humans; D007105:Immune Complex Diseases / Q000209:etiology / Q000378:metabolism*; D007074:Immunoglobulin G / Q000302:isolation & purification / Q000378:metabolism; D007075:Immunoglobulin M / Q000302:isolation & purification / Q000378:metabolism*; D007145:Immunoglobulin kappa-Chains / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D012217:Rheumatoid Factor / Q000378:metabolism*; D018709:Statistics, Nonparametric; D008258:Waldenstrom Macroglobulinemia / Q000378:metabolism
D001323:Autoantibodies; D003450:Cryoglobulins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007145:Immunoglobulin kappa-Chains; D012217:Rheumatoid Factor