Expression of novel secreted isoforms of human immunoglobulin E proteins.
Four human IgE isoforms produced by alternative splicing of the epsilon primary transcript were expressed as chimeric mouse/human anti 5-dimethylamino-1-naphthalenesulfonyl antibodies in the murine myeloma cell line Sp2/0. The four isoforms include the classic secreted form and three novel isoforms with altered carboxyl termini. All of these isoforms lack the transmembrane region encoded by the M1/M1' exon and are therefore predicted to be secreted proteins. When expressed in Sp2/0 cells, three of the IgE isoforms are assembled into complete molecules of two Ig heavy chains and two Ig light chains, whereas the fourth isoform is predominately assembled into half-molecules of one Ig heavy chain and one Ig light chain. All four isoforms are secreted with similar kinetics. In contrast, when the isoform containing the C epsilon4 domain joined directly to the M2 exon (IgE grandé) is expressed in the J558L cell line, it is degraded intracellularly, suggesting a cell line-dependent regulation of secretion. These data show that these novel isoforms of human IgE, predicted to occur from in vivo and in vitro mRNA analysis, can be produced and secreted by mammalian cells. The different forms of IgE may have physiologically relevant but distinct roles in human IgE-mediated immune inflammation. The availability of purified recombinant human IgE isoforms makes it possible to analyze the functional differences among them.
Lyczak
J B
JB
Department of Microbiology and Molecular Genetics, School of Medicine, University of California, Los Angeles, California 90095, USA.
Zhang
K
K
Saxon
A
A
Morrison
S L
SL
D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007073:Immunoglobulin E / Q000096:biosynthesis* / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D051379:Mice; D009101:Multiple Myeloma; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D015183:Restriction Mapping; D013462:Sulfur Radioisotopes; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013462:Sulfur Radioisotopes; D007073:Immunoglobulin E; D008715:Methionine