Comparison of IgE expression at the mRNA and protein levels in vitro.
The regulating effects of IL-4 and pokeweed mitogen on IgE synthesis in vitro by human peripheral blood leucocytes has been compared with the corresponding effect of these regulators on the expression of IgE mRNA. The latter was measured by dot blot hybridization with an oligonucleotide coding for a unique six amino acid region of the CH epsilon 2 domain. Specificity of the oligonucleotide probe was established by its inability to hybridize with RNA extracted from HMY-2 (IgG) and XQ-15 (IgM) secreting cell lines whilst producing intense signals with RNA extracted from the IgE secreting cell line U266. Whilst IgE mRNA was detected in RNA extracted from PBL of both atopic and control subjects, spontaneous IgE synthesis was restricted to atopic PBL. IL-4 increased both IgE mRNA and IgE synthesis in all PBL samples but PWM, while significantly increasing IgE mRNA expression either failed to modify IgE synthesis or actively suppressed it. The assay system employed to quantitate IgE synthesis in vitro was shown to be inhibited by both IgE binding factors and IgG anti-IgE autoantibodies which are produced in PBL cultures. IgE mRNA levels might therefore more accurately monitor the regulatory effects of IL-4 and PWM on IgE synthesis than quantitation of the IgE by radioimmunoassay.
Turner
K J
KJ
Department of Microbiology, University of Western Australia, Medical Centre.
Creany
J
J
Coelen
R J
RJ
Cameron
K J
KJ
Holt
B J
BJ
Beilharz
M W
MW
D002460:Cell Line; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics*; D015847:Interleukin-4 / Q000276:immunology; D007962:Leukocytes / Q000276:immunology; D015345:Oligonucleotide Probes; D011043:Pokeweed Mitogens / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis*; D011863:Radioimmunoassay
D015345:Oligonucleotide Probes; D011043:Pokeweed Mitogens; D012333:RNA, Messenger; D015847:Interleukin-4; D007073:Immunoglobulin E