Characterization of Fc epsilonRI expressing human monocytic cell lines. 1. The role of CD45 on signal transduction in primary monocytes and cell lines.
BACKGROUND
Recently, the high affinity receptor for IgE (Fc epsilonRI), which plays a major role in allergies, has been identified on a number of different antigen-presenting cell types, including human monocytes from atopic and nonatopic donors. In this report human monocytic cell lines were used to test for the expression of Fc epsilonRI, reasoning that a monocytic cell line expressing Fc epsilonRI constitutively would be a useful tool for large scale studies on the regulation of IgE binding and signal transduction.


METHODS
Reverse transduction polymerase chain reaction was applied to identify Fc epsilonRI alpha-chain message, flow cytometry to detect Fc epsilonRI surface expression and signal transduction on the cell lines generated by transfection.


RESULTS
We report the establishment of monocytic cell lines constitutively expressing Fc epsilonRI (THP1-alpha01 to THP1-alpha40) generated by transfection of the cell line THP1 with a plasmid encoding the Fc epsilonRI alpha-chain only. Fc epsilonRI on the THP1-alpha lines specifically binds IgE and is functional with regard to ligand binding and signal transduction. Comparative studies between the transfectants and primary human monocytes from nonatopic donors demonstrated the regulatory role of the tyrosine phosphatase CD45 on Fc epsilonRI-mediated cell activation.


CONCLUSIONS
Monocytic cell lines carry Fc epsilonRI alpha-chain RNA and enhancement by transfection results in surface Fc epsilonRI expression on THP1. Triggering the receptor on the THP1-alpha lines or on human monocytes, which express native Fc epsilonRI, elicits a rapid and transient calcium mobilization, prevented by co-cross-linking of Fc epsilonRI and CD45.
Reischl
I G
IG
Department of Immunodermatology, Sandoz Research Institute, Vienna, Austria.
Corvaïa
N
N
Unger
J
J
Woisetschläger
M
M
Mudde
G C
GC
D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000235:genetics / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000276:immunology / Q000502:physiology; D009000:Monocytes / Q000276:immunology*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D017455:Receptors, IgE / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D017455:Receptors, IgE; D007073:Immunoglobulin E; D017493:Leukocyte Common Antigens; D002118:Calcium