Expression of recombinant transmembrane CD59 in paroxysmal nocturnal hemoglobinuria B cells confers resistance to human complement
| Autor: | R P, Rother, S A, Rollins, J, Mennone, A, Chodera, S A, Fidel, M, Bessler, P, Hillmen, S P, Squinto |
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| Rok vydání: | 1994 |
| Předmět: |
B-Lymphocytes
Herpesvirus 4 Human Mice Inbred BALB C Membrane Glycoproteins Base Sequence Glycosylphosphatidylinositols Molecular Sequence Data Gene Transfer Techniques Hemoglobinuria Paroxysmal Gene Expression CD59 Antigens 3T3 Cells Complement System Proteins Recombinant Proteins Cell Line Mice L Cells Antigens CD Animals Humans Cell Line Transformed |
| Zdroj: | Blood. 84(8) |
| ISSN: | 0006-4971 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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