CD32A (Fc gamma RIIa) mRNA expression and regulation in blood monocytes and cell lines.
The cell surface expression of the CD32 receptors for the Fc portion of immunoglobulin G (Fc gamma RII) is highly regulated by agents such as phorbol ester (PMA) and cytokines. In this study we investigated the regulatory effects of PMA, aggregated IgG (AIgG) and KuFc79 anti-CD32 monoclonal antibodies (mAb) on the expression of the CD32A isomer mRNA. When U937 (CD32+ cells) are incubated with PMA a significant enhancement of the CD32A isomer mRNA is observed. The CD32A mRNA is also markedly enhanced when the CD32+ K562 cells are incubated with AIgG and anti-CD32 mAb but not with control MOPC-195 mAb. The addition of actinomycin D (ActD), a transcriptional inhibitor together with PMA, AIgG or KuFc79 mAb diminishes the enhanced levels of CD32A mRNA to the basal, constitutively expressed levels, implicating transcriptional regulation in this modulatory process. The PMA induced mRNA is rapidly degraded while the constitutively expressed CD32A mRNA is not, suggesting differential regulation of the stimulated vs the unstimulated CD32A mRNA. Inhibition of protein synthesis does not significantly affect the upregulation of CD32 mRNA by PMA, AIgG or KuFc79 in U937 and K562 cells. The upregulation of CD32A mRNA observed in the cell lines U937 and K562 is also detected when normal blood monocytes are used. Similarly to the cell lines the enhancement of CD32A mRNA in monocytes is blocked by ActD.
Alevy
Y G
YG
Department of Surgery, Washington University School of Medicine, St Louis, MO 63110.
Tucker
J
J
Mohanakumar
T
T
D000293:Adolescent; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D015342:DNA Probes; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D008969:Molecular Sequence Data; D009000:Monocytes / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000302:isolation & purification; D017452:Receptors, IgG / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D000911:Antibodies, Monoclonal; D015342:DNA Probes; D007074:Immunoglobulin G; D012333:RNA, Messenger; D017452:Receptors, IgG; D003609:Dactinomycin; D003513:Cycloheximide; D013755:Tetradecanoylphorbol Acetate