Production of a Chimeric Form of CD23 That Is Oligomeric and Blocks IgE Binding to the FcεRI

Abstract

The low affinity receptor for IgE (FcεRII/CD23) has previously been shown to interact with IgE with a dual affinity. Three chimeric constructs were created containing the lectin domain (amino acids 172–188) or the “neck” and lectin domain (amino acids 157–188) attached to subunits of oligomeric proteins. All chimeras were incapable of interacting with IgE with either a high or low affinity, indicating that the α-helical stalk of CD23 is important for orienting the lectin heads such that an interaction with IgE can occur. This concept received further support in that a chimeric CD23 composed of the human CD23 stalk and the mouse CD23 lectin head bound mouse IgE with a dual affinity, but could only bind rat IgE with a low affinity. Effort was next concentrated on a construct consisting of the entire extracellular (EC) region of CD23. A mutation to the first cleavage site of CD23 (C1M) resulted in a more stable molecule as determined by a decrease of soluble CD23 release. A soluble chimeric EC-C1M was prepared by attaching an isoleucine zipper to the amino terminus (lzEC-C1M). The interaction with IgE by lzEC-C1M was found to be superior to that seen with EC-CD23. The lzEC-C1M could inhibit binding of IgE to both CD23 and the high affinity receptor for IgE, FcεRI, providing further evidence for a strong interaction with IgE. FcεRI inhibition (∼70%) was seen at equimolar concentrations of lzEC-C1M, implying the effectiveness of this chimera and suggesting its potential therapeutic value.