Expression of a recombinant murine IgE in transfected myeloma cells.
We constructed a recombinant gene encoding an immunoglobulin (Ig) heavy chain consisting of the variable region from the phosphorylcholine (PC)-specific secreting myeloma MOPC167 and the epsilon constant region from SJL mice. This gene, cloned into the shuttle vector pSV2gpt, was transfected into J558L myeloma cells, and stable transformants that expressed the epsilon gene were cloned. The IgE heavy chain in these transformants is associated with the endogenous lambda light chain and is secreted as an intact IgE molecule. However, the secreted IgE does not bind to PC conjugated to bovine serum albumin (PC-BSA). The MOPC167 kappa chain gene was cloned into the shuttle vector pSV2neo and was transfected into the epsilon heavy-chain transformant. Stable transformants were cloned that expressed both the epsilon heavy chain and the kappa light chain. IgE secreted from such a transformant was shown to bind to PC-BSA. Both types of secreted recombinant IgE bound to rat basophilic leukemia (RBL) cells, but only the IgE produced by the cell line transformed with the MOPC167 kappa gene could be cross-linked with PC-BSA to cause serotonin release.
D000818:Animals; D000918:Antibody Specificity; D002460:Cell Line; D002794:Choline / Q000031:analogs & derivatives*; D006636:Histamine Release; D007073:Immunoglobulin E / Q000235:genetics* / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007139:Immunoglobulin epsilon-Chains / Q000235:genetics*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics*; D051379:Mice; D010767:Phosphorylcholine / Q000276:immunology*; D010954:Plasmacytoma; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D007143:Immunoglobulin Heavy Chains; D007139:Immunoglobulin epsilon-Chains; D007145:Immunoglobulin kappa-Chains; D011994:Recombinant Proteins; D010767:Phosphorylcholine; D007073:Immunoglobulin E; D002794:Choline