Autoimmunity mediated by
IgG4 subclass
autoantibodies is an expanding field of research. Due to their structural characteristics a key feature of
IgG4 antibodies is the ability to exchange Fab-arms with other, unrelated,
IgG4 molecules, making the
IgG4 molecule potentially monovalent for the specific
antigen. However, whether those disease-associated
antigen-specific
IgG4 are mono- or divalent for their
antigens is unknown.
Myasthenia gravis (MG) with
antibodies to muscle specific
kinase (
MuSK-MG) is a well-recognized disease in which the predominant pathogenic
IgG4 antibody binds to extracellular
epitopes on
MuSK at the neuromuscular junction; this inhibits a pathway that clusters the
acetylcholine (
neurotransmitter) receptors and leads to failure of neuromuscular transmission. In vitro Fab-arm exchange-inducing conditions were applied to
MuSK antibodies in sera, purified
IgG4 and IgG1-3 sub-fractions. Solid-phase cross-linking assays were established to determine the extent of pre-existing and inducible Fab-arm exchange. Functional effects of the resulting populations of
IgG4 antibodies were determined by measuring inhibition of
agrin-induced AChR clustering in C2C12 cells. To confirm the results, κ/κ, λ/λ and hybrid κ/λ IgG4s were isolated and tested for
MuSK antibodies. At least fifty percent of patients had
IgG4, but not IgG1-3,
MuSK antibodies that could undergo Fab-arm exchange in vitro under reducing conditions. Also
MuSK antibodies were found in vivo that were divalent (monospecific for
MuSK). Fab-arm exchange with normal human
IgG4 did not prevent the inhibitory effect of serum derived
MuSK antibodies on AChR clustering in C2C12 mouse myotubes. The results suggest that a considerable proportion of
MuSK IgG4 could already be Fab-arm exchanged in vivo. This was confirmed by isolating endogenous
IgG4 MuSK antibodies containing both κ and λ light chains, i.e. hybrid
IgG4 molecules. These new findings demonstrate that Fab-arm exchanged
antibodies are pathogenic.