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Blood, Vol. 92 No. 5 (September 1), 1998:
pp. 1785-1792
Targeted Inactivation of Murine Band 3 (AE1) Gene Produces a
Hypercoagulable State Causing Widespread Thrombosis In Vivo
Hani Hassoun,
Ying Wang,
John Vassiliadis,
Mohini Lutchman,
Jiri Palek,
Leo Aish,
Irene S. Aish,
Shih-Chun Liu, and
Athar H. Chishti
From the Department of Biomedical Research, St Elizabeth's Medical
Center, Tufts University School of Medicine, Boston, MA.
Only 5% to 10% of band 3 null mice survive the neonatal period. To
determine the cause of death, 3 adult and 11 newborn band 3 null mice
were submitted for histopathologic examination. All but 1 pup showed
evidence of thrombosis including: (1) large thrombotic lesions in the
heart, which were partially organized, calcified in some fields, and
endothelialized, indicating a process that developed premortem (3 of 3 adults and 6 of 11 pups). (2) Subcapsular necrotic areas in the liver
suggestive of premortem ischemic events caused by arteriolar occlusions
(8 of 11 pups). (3) Large vein thrombi (4 of 11 pups). To investigate
the etiology of this hypercoagulable state, we have used the Russell's
viper venom test (RVV) to show that red blood cells (RBCs) from band 3 null mice significantly shorten the RVV clotting time of normal plasma
in a dose-dependent fashion, whereas RBCs from normal mice have no
effect, suggesting that the membrane of band 3 null RBCs provides a
suitable surface for activation of the prothrombinase complex. Using
flow cytometry, we have examined the phosphatidylserine (PS)-specific
binding of fluorescein isothiocyanate (FITC)-annexin V to normal and
band 3 null RBCs. A subpopulation of cells (3% to 5% of RBCs) with increased FITC-annexin V binding was detected in band 3 null RBCs as
compared with normal RBCs. Furthermore, the entire cell population of
band 3 null RBCs shows a measurable increase in the mean fluorescence intensity, suggesting that band 3 null RBCs may have increased PS
exposure on the outer membrane leaflet. These findings are further
supported by direct fluorescence microscopy of normal and band 3 null
RBCs labeled with FITC-annexin V. Based on these observations, we
postulate that the high mortality of band 3 null mice may be related to
a hypercoagulable state, which appears to originate from changes in the
phospholipid composition of the membrane leading to PS exposure on the
outer leaflet.
© 1998 by The American Society of Hematology.
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