21 research outputs found
A Soluble Form of the High Affinity IgE Receptor, Fc-Epsilon-RI, Circulates in Human Serum
Soluble IgE receptors are potential in vivo modulators of
IgE-mediated immune responses and are thus important for our basic understanding
of allergic responses. We here characterize a novel soluble version of the
IgE-binding alpha-chain of Fc-epsilon-RI (sFcεRI), the high affinity
receptor for IgE. sFcεRI immunoprecipitates as a protein of ∼40 kDa and
contains an intact IgE-binding site. In human serum, sFcεRI is found as a
soluble free IgE receptor as well as a complex with IgE. Using a newly
established ELISA, we show that serum sFcεRI levels correlate with serum IgE
in patients with elevated IgE. We also show that serum of individuals with
normal IgE levels can be found to contain high levels of sFcεRI. After
IgE-antigen-mediated crosslinking of surface FcεRI, we detect sFcεRI in
the exosome-depleted, soluble fraction of cell culture supernatants. We further
show that sFcεRI can block binding of IgE to FcεRI expressed at the cell
surface. In summary, we here describe the alpha-chain of FcεRI as a
circulating soluble IgE receptor isoform in human serum
IgE binds asymmetrically to its B cell receptor CD23.
The antibody IgE plays a central role in allergic disease mechanisms. Its effector functions are controlled through interactions between the Fc region and two principal cell surface receptors FcεRI and CD23. The interaction with FcεRI is primarily responsible for allergic sensitization and the inflammatory response, while IgE binding to CD23 is involved in the regulation of IgE synthesis and allergen transcytosis. Here we present the crystal structure of a CD23/IgE-Fc complex and conduct isothermal titration calorimetric binding studies. Two lectin-like "head" domains of CD23 bind to IgE-Fc with affinities that differ by more than an order of magnitude, but the crystal structure reveals only one head bound to one of the two identical heavy-chains in the asymmetrically bent IgE-Fc. These results highlight the subtle interplay between receptor binding sites in IgE-Fc and their affinities, the understanding of which may be exploited for therapeutic intervention in allergic disease
The mechanistic and functional profile of the therapeutic anti-IgE antibody ligelizumab differs from omalizumab.
Targeting of immunoglobulin E (IgE) represents an interesting approach for the treatment of allergic disorders. A high-affinity monoclonal anti-IgE antibody, ligelizumab, has recently been developed to overcome some of the limitations associated with the clinical use of the therapeutic anti-IgE antibody, omalizumab. Here, we determine the molecular binding profile and functional modes-of-action of ligelizumab. We solve the crystal structure of ligelizumab bound to IgE, and report epitope differences between ligelizumab and omalizumab that contribute to their qualitatively distinct IgE-receptor inhibition profiles. While ligelizumab shows superior inhibition of IgE binding to FcεRI, basophil activation, IgE production by B cells and passive systemic anaphylaxis in an in vivo mouse model, ligelizumab is less potent in inhibiting IgE:CD23 interactions than omalizumab. Our data thus provide a structural and mechanistic foundation for understanding the efficient suppression of FcεRI-dependent allergic reactions by ligelizumab in vitro as well as in vivo
The crystal structure of IgE Fc reveals an asymmetrically bent antibody conformation
The distinguishing structural feature of immunoglobulin E (IgE), the antibody responsible for allergic hypersensitivity, is the C epsilon 2 domain pair that replaces the hinge region of IgG. The crystal structure of the IgE Fc (constant fragment) at a 2.6-A resolution has revealed these domains. They display a distinctive, disulfide-linked Ig domain interface and are folded back asymmetrically onto the C epsilon 3 and C epsilon 4 domains, which causes an acute bend in the IgE molecule. The structure implies that a substantial conformational change involving C epsilon 2 must accompany binding to the mast cell receptor Fc epsilon RI. This may be the basis of the exceptionally slow dissociation rate of the IgE-Fc epsilon RI complex and, thus, of the ability of IgE to cause persistent allergic sensitization of mast cells