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Granulocyte-macrophage colony-stimulating factor and interleukin-3 mRNAs
are produced by a small fraction of blood mononuclear cells
JZ Wimperis, CM Niemeyer, CA Sieff, B Mathey-Prevot, DG Nathan and RJ Arceci
Division of Pediatric Oncology and Hematology, Dana-Farber Cancer
Institute, Boston.
Northern blot analysis has identified granulocyte macrophage colony
stimulating factor (GM-CSF) mRNA in monocytes and both GM-CSF and
interleukin-3 (IL-3) mRNA in lymphocytes. However, these results have not
addressed whether all cells or a subset of the population is capable of
hematopoietic growth factor (HGF) production. To resolve this question, we
applied in situ hybridization of radiolabeled antisense RNA probes to
centrifuged preparations of total blood mononuclear cells (BMCs) and
fractionated lymphocyte subpopulations. Without stimulation, no circulating
cells expressed detectable levels of GM-CSF or IL-3 mRNA. On stimulation of
BMCs with phorbol myristate acetate (PMA) and phytohemagglutinin or PMA and
the calcium ionophore ionomycin, approximately 5% expressed GM-CSF mRNA and
approximately 1% IL-3 mRNA. Control sense probes produced no labeled cells.
To determine the subsets of lymphocytes capable of GM-CSF and IL-3
expression, BMCs were fractionated by FACS into CD8+ and CD4+ lymphocyte
subsets and CD16+ (NK) cells. The unfractionated cells and cell fractions
were then stimulated with PMA and ionomycin. Results demonstrated that 3%
to 5% of the CD16+, CD8+, and CD4+ lymphocytes produced GM-CSF mRNA.
However, the number of IL-3 mRNA-positive cells in the FACS-sorted subsets
was greatly reduced (0.02% to 0.05%) as compared with the unseparated cells
(1%). Treatment of BMCs with high-dose interleukin-2 (IL-2) for 1 week
followed by PMA plus ionomycin resulted in a lymphocyte population in which
50% and 3% of cells expressed GM-CSF and IL-3 mRNA, respectively. Thus,
GM-CSF and IL-3 mRNA expression in T cells and NK cells is restricted to a
small fraction of cells that can be greatly expanded by IL-2 stimulation.
These results suggest a possible physiologic mechanism for increasing HGF
production by circulating lymphocytes.
Volume 74,
Issue 5,
pp. 1525-1530,
10/01/1989
Copyright © 1989 by The American Society of Hematology
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