Post-transcriptional regulation of interferon-alpha 4 subtype production by lymphoblastoid cells.
The constitutive production of interferon-alpha (IFN-alpha) subtypes by the lymphoblastoid cell lines, Namalwa, Daudi and Raji, was investigated using sensitive and semi-quantitative flow cytometric techniques. Further, we sought to determine whether the previously described failure of these cell lines to produce IFN-alpha-4 was a result of the deletion of the IFN A4 gene. Cytoplasmic production of IFN-alpha-2 and IFN-alpha-4 was assessed using IFN-alpha subtype-specific antipeptide antibodies and FITC-labelled secondary antibodies in indirect immunofluorescence-flow cytometry studies. The constitutive production of IFN-alpha-2 was detected in all three cell lines. Significant increases in fluorescence representing increased production of IFN-alpha-2 and possibly other IFN-alpha subtypes were detected after induction by Sendai virus. Approximately 100 per cent of cells in the Namalwa, Daudi and Raji cell populations contained IFN-alpha-2 before and after induction. However, no cells from the same cell populations contained the IFN-alpha-4 subtype. Analysis of genomic DNA isolated from the lymphoblastoid cells using the Polymerase Chain Reaction (PCR) and oligonucleotide primers specific for IFN A2 or IFN A4 confirmed the presence of the genes encoding both IFN-alpha subtypes. Furthermore, using reverse transcriptase-PCR amplification, mRNAs for both IFN-alpha-2 and IFN-alpha-4 were detected. Therefore, in contrast to some leukaemias and derived cell lines where IFN A genes have been deleted, these cell lines of B cell lineage exhibit selective expression of IFN A genes, as a result of altered transcriptional/translational control of IFN-alpha expression.
Greenway
A L
AL
Centre for Molecular Biology and Medicine, Monash University, Clayton, Victoria, Australia.
Brandt
E R
ER
Hertzog
P J
PJ
Devenish
R J
RJ
Linnane
A W
AW
D000998:Antiviral Agents / Q000494:pharmacology; D002478:Cells, Cultured; D003470:Culture Media; D003593:Cytoplasm / Q000378:metabolism; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D008214:Lymphocytes / Q000378:metabolism* / Q000502:physiology; D046911:Macromolecular Substances; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis / Q000235:genetics; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism
D000998:Antiviral Agents; D003470:Culture Media; D016898:Interferon-alpha; D046911:Macromolecular Substances; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase