Potentiating and suppressive IgE-binding factors are expressed by a single cloned gene.
We have investigated expression of an IgE-binding factor (IgE-BF) cDNA in both COS-7 monkey kidney cells and Chinese hamster ovary cells. Transient expression of the IgE-BF clone in either cell type yielded IgE-BF, which potentiated an in vitro IgE response and had an affinity for lentil lectin. In contrast, when the transient expression experiments were carried out in the presence of tunicamycin, the factors no longer bound to lentil lectin. Moreover, IgE-BF expressed under these conditions suppressed an in vitro IgE response. IgE-BF lacking affinity for lentil lectin and suppressing the IgE response also resulted from transient expression of the IgE-BF gene in the presence of glycosylation inhibiting factor, a phospholipase inhibitory protein. Thus, IgE-BF that either potentiate or suppress the IgE response can be expressed from a single cloned gene; the difference in biological activities appears to be determined principally by the type of glycosylation of the common polypeptide chain. Previous work showed that IgE-BF bears an antigenic determinant recognized by the anti-Ia monoclonal antibody OX3. IgE-BF produced in the presence of tunicamycin, and IgE-BF expressed from a mutant cDNA lacking one of two carbohydrate-attachment sites, lacked the OX3 determinant. Thus, the OX3 determinant on IgE-BF appears to be associated with a site of N-linked glycosylation.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D005796:Genes / Q000187:drug effects; D007073:Immunoglobulin E / Q000276:immunology; D008222:Lymphokines / Q000235:genetics*; D009154:Mutation; D029584:Prostatic Secretory Proteins; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D008222:Lymphokines; D029584:Prostatic Secretory Proteins; C043076:beta-microseminoprotein; C013059:immunoglobulin-binding factors; D014415:Tunicamycin; D007073:Immunoglobulin E; D004262:DNA Restriction Enzymes