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Department of Pathology, University of Maryland, School of Medicine, Baltimore, MD 21201
We have previously shown that generation of sublytic C5b-9, the
membrane attack complex of complement, induces oligodendrocytes to
enter cell cycle and reduces apoptotic cell death in vitro. In the
present study, the cellular factors involved in apoptosis of
oligodendrocyte progenitor cells and oligodendrocytes, and the
inhibitory effect of C5b-9 on apoptotic process were investigated.
Oligodendrocyte progenitor cells identified by mAb A2B5 that were
isolated from neonatal rat brains were differentiated into
oligodendrocytes in serum-free defined medium. The differentiation,
which occurs simultaneously with apoptotic cell death, was associated
with a rapid loss of bcl-2 mRNA and increased expression
of caspase-3 mRNA. Activation of caspase-3 in differentiating cells was
demonstrated by the generation of 17- and 12-kDa fragments of caspase-3
proenzyme and by cleavage of poly(ADP-ribose) polymerase, a specific
caspase-3 substrate. Cell death associated with differentiation was
inhibited by the caspase-3 inhibitor DEVD-CHO in a dose-dependent
manner. Assembly of sublytic C5b-9 resulted in inhibition of caspase-3
activation. In addition, synthesis of BCL-2 protein in oligodendrocytes
was significantly increased by C5b-9. The TNF-
-induced apoptosis of
oligodendrocytes was also inhibited by C5b-9. These results indicate
that up-regulation of BCL-2 protein and inhibition of caspase-3
activation are potential mechanisms by which C5b-9 increases survival
of oligodendrocyte in vitro and possibly in vivo during inflammation
and immune-mediated demyelination affecting the
CNS.
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