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Interferon-gamma gene expression in human B-cell lines: induction by
interleukin-2, protein kinase C activators, and possible effect of
hypomethylation on gene regulation
Y Pang, Y Norihisa, D Benjamin, RR Kantor and HA Young
Laboratory of Experimental Immunology, BRMP, NCI-FCRDC, Frederick, MD
21702-1201.
Human interferon-gamma (IFN-gamma) is an important immunomodulatory protein
produced predominantly by T cells and large granular lymphocytes (LGLs).
Whereas large amounts of data have been accumulated regarding IFN gamma
gene expression in these two cell types, little information about IFN gamma
expression in other cell types exists. In this study, we have analyzed the
production of IFN gamma by the Epstein- Barr virus (EBV)-positive B-cell
line, JLP(c), derived from a patient with Burkitt's lymphoma, and another
human B-cell line, PA682BM-1, which was derived from an acquired
immunodeficiency syndrome patient. Southern blot analysis indicates the
presence of an Ig heavy chain gene rearrangement, but no rearrangement of
the T-cell receptor beta chain gene or IFN gamma gene in these B-cell
lines. Both cell lines were found to express surface IgD and other B-cell
surface markers, thus confirming their B-cell lineage. Analysis for surface
Ig, cytoplasmic Ig, and secreted Ig indicates that the two cell lines are
in relatively early stages of the B-cell differentiation pathway. We now
report that PA682BM-1 can be triggered by the protein kinase C (PKC)
activators, phorbol 12-myristate 13-acetate (PMA) and (-)Indolactam-v, to
secrete IFN gamma, whereas JLP(c) cells spontaneously produce low levels of
IFN gamma that can be enhanced by PKC activators and interleukin-2 (IL-2).
After activation of the cell lines with IL-2, (-)Indolactam-v, and PMA,
increases in cytoplasmic messenger RNAs (mRNAs) of IFN gamma and the IL- 2
receptor chains were also observed. The induction of IFN gamma mRNA and
protein by IL-2 was completely blocked by a monoclonal antibody to IL-2
receptor p75 (beta chain), but not by the monoclonal antibody to p55 (alpha
chain). Analysis of IFN gamma genomic DNA indicates that the gene is not
amplified, but that hypomethylation in the 5' noncoding region of the IFN
gamma gene has occurred in the B-cell line from the Burkitt's lymphoma
patient that spontaneously produces IFN gamma. This finding suggests that
the methylation state of the promoter region may play an important role in
the control of IFN gamma gene expression in B cells.
Volume 80,
Issue 3,
pp. 724-732,
08/01/1992
Copyright © 1992 by The American Society of Hematology
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