Density ultracentrifugation of sickle cells during and after pain crisis:
increased dense echinocytes in crisis
JA Warth and DL Rucknagel
An increase in the number of irreversibly sickled cells (ISCs) in pain
crisis has been found by some investigators but not others. We have used
the technique of discontinuous arabinogalactan density-gradient
ultracentrifugation of whole blood to study ISCs from patients with sickle
cell anemia (SCA) during pain crisis and again when pain free (5- 331 days
after crisis). Nine patients have been studied through ten episodes of pain
crisis. Five layers with densities from 1.128 g/mL to 1.158 g/mL have been
used. Careful classification of the cells using Nomarsky optics
demonstrated highly significant changes occurring in the layers of the
gradient. The changes involve the appearance of an increased percentage of
echinocytic ISCs and echinocytic cells that were not ISCs, especially in
the denser gradient layers, during crisis, and their replacement by
normal-appearing discocytes in the pain-free state. There was no change in
ISCs that were not echinocytic. Data collected previously demonstrated that
reduced glutathione activity correlated with increased echinocytosis in our
gradient layers. This indicates that the echinocytic change may occur as a
result of oxidant stress. Hemoglobin F levels and the percentage of
hemoglobin F from reticulocytes showed no consistent change to coincide
with the rise in normal-appearing discocytes in the lightest layers after
crisis. Our data indicate that pain crisis occurs in association with an
echinocytic change, which may be induced by oxidant injury. The rise in
normal-appearing cells after crisis may reflect increased hemoglobin F
production in some patients but mainly relates to the disappearance of
these echinocytic erythrocytes.
Volume 64,
Issue 2,
pp. 507-515,
08/01/1984
Copyright © 1984 by The American Society of Hematology
