Control of IFN-inducible MxA gene expression in human cells.
MxA is an IFN-induced human protein which is located in the cytoplasm of induced cells. MxA makes the cells resistant to infection by influenza and vesicular stomatitis viruses. In the present work we used baculovirus expression system to produce MxA protein. The protein was purified to homogeneity and highly specific polyclonal anti-MxA antibodies were prepared. In human mononuclear cells, and A549 lung carcinoma cells expression of MxA protein is induced by very low (< 1 IU/ml) doses of leukocyte IFN-alpha (nIFN-alpha), whereas IFN-gamma does not seem to induce it or potentiate the induction by IFN-alpha. In mononuclear cells stimulated with high doses of leukocyte IFN-alpha concentrations, the amount of MxA mRNA was induced 10-fold at 4 h after IFN induction and up to 10-fold higher MxA protein levels were observed at 24-48 h postinduction. The gene can be reinduced by IFN-alpha 24 h after the initial induction suggesting for a lack of negative feedback after this time point. The protein is very stable, the half-life being approximately 2.3 days. Flow cytometric analysis revealed that monocytes have higher basal and induced MxA protein levels than lymphocytes but the dose-dependency of MxA expression is very similar in both cell types. Granulocytes are producing very low amounts of MxA protein.
Ronni
T
T
Molecular Biology Programme, National Public Health Institute, Helsinki, Finland.
Melén
K
K
Malygin
A
A
Julkunen
I
I
D000818:Animals; D000998:Antiviral Agents / Q000235:genetics*; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D019204:GTP-Binding Proteins; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007372:Interferons / Q000494:pharmacology*; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins; D011506:Proteins / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits
D000998:Antiviral Agents; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D011506:Proteins; D012333:RNA, Messenger; D007372:Interferons; D019204:GTP-Binding Proteins