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Expression of recombinant transmembrane CD59 in paroxysmal nocturnal
hemoglobinuria B cells confers resistance to human complement
RP Rother, SA Rollins, J Mennone, A Chodera, SA Fidel, M Bessler, P Hillmen and SP Squinto
Alexion Pharmaceuticals Inc, New Haven, CT.
Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hematopoietic
disorder characterized by complement-mediated hemolytic anemia,
pancytopenia, and venous thrombosis. These clinical manifestations arise
from an underlying molecular defect of bone marrow stem cells.
Specifically, somatic mutations in the phosphatidylinositol glycan class A
gene result in the ability of blood cells to anchor complement- regulatory
proteins (CD59 and DAF) to the cell surface via glycosyl
phosphatidylinositol (GPI). In an attempt to circumvent the functional
defect in PNH cells, a recombinant transmembrane form of CD59 (CD59-TM) was
analyzed for the ability to regulate complement activity. Balb/3T3 stable
transfectants expressing similar levels of either CD59-TM or native CD59
(CD59-GPI) were equally protected against human complement- mediated
membrane damage. Treatment of these cells with
phosphatidylinositol-specific phospholipase C failed to release CD59-TM
from the cell surface. Retroviral transduction of GPI-anchoring deficient
mouse L cells with CD59-TM resulted in surface expression of the protein
and rendered these cells resistant to human complement- mediated membrane
damage. Conversely, L cells transduced with CD59-GPI failed to express this
protein on the cell surface. A GPI-anchoring deficient complement-sensitive
B-cell line derived from a PNH patient was successfully transduced with
CD59-TM, resulting in surface expression of the protein. The PNH B cells
expressing CD59-TM were protected against classical complement-mediated
membrane damage by human serum. Taken together, these data establish that a
functional recombinant transmembrane form of CD59 can be expressed on the
surface of GPI-anchoring deficient PNH cells and suggest that retroviral
gene therapy with this molecule could provide a treatment for PNH patients.
Volume 84,
Issue 8,
pp. 2604-2611,
10/15/1994
Copyright © 1994 by The American Society of Hematology
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