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Blood, 15 April 2002, Vol. 99, No. 8, pp. 2786-2793
HEMATOPOIESIS
Hematopoietic cells expressing the peripheral cannabinoid
receptor migrate in response to the endocannabinoid
2-arachidonoylglycerol
Meritxell Alberich Jordà,
Sandra E. Verbakel,
Peter
J. M. Valk,
Yolanda V. Vankan-Berkhoudt,
Mauro Maccarrone,
Allessandro Finazzi-Agrò,
Bob Löwenberg, and
Ruud Delwel
From the Institute for Hematology, Erasmus University
Rotterdam, The Netherlands; and Department of Experimental Medicine and
Biochemical Sciences, University of Rome "Tor Vergata," Rome,
Italy.
Cb2 is a novel protooncogene encoding the peripheral
cannabinoid receptor. Previous studies demonstrated that 2 distinct
noncoding first exons exist: exon-1A and exon-1B, which both splice to
protein-coding exon-2. We demonstrate that in retrovirally induced
murine myeloid leukemia cells with proviral insertion in
Cb2, exon-1B/exon-2 Cb2 messenger RNA levels
have been increased, resulting in high receptor numbers. In myeloid
leukemia cells without virus insertion in this locus, low levels of
only exon-1A/exon-2 Cb2 transcripts were present and
receptors could not be detected. To elucidate the function of Cb2 in
myeloid leukemia cells, a set of in vitro experiments was carried out
using 32D/G-CSF-R (granulocyte colony-stimulating factor receptor)
cells transfected with exon-1B/exon-2 Cb2 complementary DNA
and a myeloid cell line carrying a virus insertion in Cb2 (ie, NFS 78). We demonstrate that a major function of the Cb2 receptor
is stimulation of migration as determined in a transwell assay.
Exposure of Cb2-expressing cells to different cannabinoids showed that
the true ligand for Cb2 is 2-arachidonoylglycerol (2-AG), which may act
as chemoatractant and as a chemokinetic agent. Furthermore, we observed
a significant synergistic activity between 2-AG and interleukin-3 or
G-CSF, suggesting cross-talk between the different receptor systems.
Radioactive-ligand binding studies revealed significant numbers of Cb2
receptors in normal spleen. Transwell experiments carried out with
normal mouse spleen cells showed 2-AG-induced migration of B220-,
CD19-, immunoglobulin M-, and immunoglobulin D-expressing B
lymphocytes. Our study demonstrates that a major function of Cb2
receptor expressed on myeloid leukemia cells or normal splenocytes is
stimulation of migration.
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