Studies of the specificity and cross-reactions of antibodies to lipid A found in juvenile arthritis

This work was started to determine whether the immunoglobulin G (IgG) reactions with monophosphoryl lipid A (MPL) found in children with arthritis were due to contaminants, a specific site on lipid A, or polyspecific binding. Different lots of MPL were examined by electrophoresis and immunoblot. Competitive inhibition of enzyme-linked immunosorbent assays (ELISAs) by analogs of MPL and biologic materials of clinical interest was used to determine the specificity of the binding site and potential cross-reactions. IgG in all patient sera tested reacted with a single band just < 6.5 kDa on immunoblots of all lots of MPL tested. The ELISAs were inhibited best by analogs of lipid A with an exposed diglucosamine core and intact polar domains. The anti-MPL was also inhibited by fetal bovine collagen types I and II and in some instances by cardiolipin, but not by keratan sulfate, proteoglycan, or DnaK heat shock protein. Lot variation was a persistent technical problem, but no protein contaminant could be demonstrated in any lot. The ELISA and immunoblot results confirmed each other. Immunoblots detected a single band of MPL reactive with IgG. This antibody remains of interest because of its disease association and correlations and because it cross-reacts with collagen and cardiolipin.

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