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Expression of interleukin-1 beta gene in candidate human hematopoietic stem
cells
K Watari, PM Lansdorp, W Dragowska, H Mayani and JW Schrader
Biomedical Research Centre, University of British Columbia, Vancouver,
Canada.
Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect
interleukin-1 beta (IL-1 beta) mRNA in candidate human hematopoietic stem
cells. The cells, obtained from adult bone marrow (BM) or umbilical cord
blood, had a CD34+ CD45RAlo CD71lo phenotype and were further fractionated
into CD38+ and CD38- or Thy-1+ and Thy-1- subpopulations. The purity of
these fractions was always more than 99%. IL-1 beta and CD34 mRNA were
detected in pools of 30 BM-derived CD34+ CD45RAlo CD71lo cells. To further
exclude any contribution by contaminating cells, individual cells were
analyzed for CD34 and IL-1 beta mRNA. Positive results were obtained with 2
of 5 individual BM- derived CD34+ CD45RAlo CD71lo CD38+ cells isolated by
micromanipulation after overnight culture in serum-free medium without any
exogenous cytokines, and 1 of 10 individual CD34+ CD45RAlo CD71lo CD38-
cells isolated immediately after sorting. Moreover, of 10 pools of three
BM- derived CD34+ CD45RAlo CD71lo cells cultured overnight in the presence
of a mixture of various cytokines (Steel factor, IL-3, IL-6, macrophage
colony-stimulating factor [M-CSF], erythropoietin, and IL-3/granulocyte-
macrophage colony-stimulating factor [GM-CSF] fusion protein), 5 were
positive for IL-1 beta mRNA. This result was compatible with more than 20%
(95% confidence limit 0.06-0.61) of the BM cells with the CD34+ CD45RAlo
CD71lo phenotype expressing IL-1 beta mRNA. IL-1 beta expression was also
consistently observed from day 0 to day 9 in liquid cultures of
cord-blood-derived CD34+ CD45RAlo CD71lo Thy-1+ or Thy-1- cells. The
cultures contained the same combination of cytokines and resulted in an
expansion of cell numbers of up to 400-fold. GM-CSF mRNA was not detected
in the equivalent of 75 cells at any day, even though it could be detected
with high sensitivity in control stromal cells. Because IL-1 beta is a
powerful and pleiotropic biomodulator of cytokines and adhesion molecules,
our observations suggest that at least some primitive hematopoietic cells
do not merely respond passively to signals from their environment, but may
themselves regulate the paracrine production of cytokines from neighboring
stromal cells.(ABSTRACT TRUNCATED AT 400 WORDS)
Volume 84,
Issue 1,
pp. 36-43,
07/01/1994
Copyright © 1994 by The American Society of Hematology
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