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Blood, 1 January 2002, Vol. 99, No. 1, pp. 357-363
RED CELLS
Modulation of Gardos channel activity by cytokines in sickle
erythrocytes
Alicia Rivera,
Petr Jarolim, and
Carlo Brugnara
From the Department of Laboratory Medicine, Children's
Hospital Boston, and the Department of Pathology, Brigham and Women's
Hospital, Boston, MA.
It has recently been shown that the Gardos channel activity of
mouse erythrocytes can be modified by endothelins, suggesting a
functional linkage between endothelin receptors and the Gardos channel.
Using 86Rubidium (86Rb) influx, effects
were estimated of proinflammatory molecules such as platelet activator
factor (PAF), endothelin-1 (ET-1), interleukin-10 (IL-10), and
regulated on activation normal T cells expressed and secreted (RANTES)
on the Gardos channel activity in human normal and sickle red cells. It
was found that PAF (EC50: 15 ± 7 nM), RANTES
(EC50, 9 ± 6 ng/mL [1.2 ± 0.8 nM]), IL-10
(EC50, 11 ± 8 ng/mL [204 ± 148 nM]), and ET-1
(EC50, 123 ± 34 nM) induce a significant increase in
Gardos channel activity between 28% and 84%over the control. In
addition, these agents modify the Gardos channel affinity for internal
Ca++ (K0.5) by 2- to 6-fold. Biochemical
evidence is provided for the presence of ET receptor subtype B in
sickle and normal red cells. Furthermore, it was found that ET-1, PAF,
RANTES, and IL-10 induce a significant increase in red cell density
(P < .05). These data suggest that activation of the
Gardos channel is functionally coupled to receptor motifs such as C-X-C
(PAF), C-C (RANTES), and ET receptor subtype B. Thus, cell volume
regulation or erythrocyte hydration states might be altered by
activation of the Gardos channel by cytokines in vivo. The role of
these mediators in promoting sickle cell dehydration in vivo is under investigation.
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