Decreased in vivo survival of hydrogen peroxide-damaged baboon red blood
cells
J McKenney, CR Valeri, N Mohandas, N Fortier, A Giorgio and LM Snyder
Division of Hematology, St Vincent Hospital, Worcester, MA 01604.
In this study we attempt to establish the consequence of in vitro hydrogen
peroxide (H2O2)-induced membrane damage as manifested by
spectrin-hemoglobin (Sp-Hb) complex formation and decreased red blood cell
(RBC) deformability to in vivo RBC survival in baboons. After exposure to
135 to 581 mumols/L H2O2 and reduction with dithiothreitol (DTE), baboon
RBCs were infused into the animal, and the fraction of cells remaining in
circulation after 24 hours and the lifespan of surviving cells were
quantitated. In a dose-dependent fashion, a positive correlation was
observed between in vitro membrane alterations and the 24-hour in vivo
survival. While 12% of the control cells were removed from circulation in
24 hours, 23% were removed after treatment with 339 mumols/L H2O2, and 36%
following exposure to 581 mumols/L H2O2. Pretreatment with carbon monoxide
before exposure with H2O2 increased the survival of oxidized RBCs. RBCs not
removed from circulation in the first 24 hours had a normal lifespan.
Moreover, by selectively isolating biotin-labeled, peroxide-treated cells
that survived the first 24-hour posttransfusion period, a significant
decrease in Sp-Hb crosslinking was observed in these cells. These results
suggest that a subpopulation of cells sensitive to oxidation were removed
during the first 24 hours. To identify this population, the survival of
density-fractionated RBCs exposed to oxidant stress was quantitated. No
differences in either the 24-hour survival or RBC life span were observed
between untreated low-density (MCHC less than or equal to 32g/dL) and
high-density cells (MCHC greater than or equal to 37g/dL). However,
striking differences were noted after treatment with 339 mumols/L H2O2,
with the 24-hour survival of high-density cells showing a marked decrease
compared with low-density cells. These data support our hypothesis that
during peroxidative membrane damage, Hb oxidation initiates a sequence of
events resulting in skeletal changes that lead to membrane alterations and,
eventually, in vivo destruction, and that the dense, dehydrated cells are
more susceptible to oxidant damage.
Volume 76,
Issue 1,
pp. 206-211,
07/01/1990
Copyright © 1990 by The American Society of Hematology
