title: The population of paroxysmal nocturnal hemoglobinuria neutrophils deficient in decay-accelerating factor is also deficient in alkaline phosphatase.
abstract: In patients with paroxysmal nocturnal hemoglobinuria (PNH) the RBCs, neutrophils (PMNs), monocytes, and platelets derived from the abnormal clone are deficient in the complement-regulatory protein decay-accelerating factor (DAF). RBC acetylcholinesterase (AChE) and leukocyte alkaline phosphatase (LAP) activities are also characteristically low. DAF, AChE, and LAP are known to be anchored within cell membranes to glycophospholipid-containing phosphatidylinositol (PI). Because PNH progenitors contain DAF that appears to be lost with maturation, it has been proposed that this disorder results from abnormal tethering of these and possibly other proteins to membrane PI. We were puzzled, therefore, that our two PNH patients consistently had normal LAP levels. Consequently, we studied their isolated PMNs to compare DAF and LAP activities in individual cells. PMNs were separated by flow cytometry into DAF-positive and -negative populations by using rabbit anti-DAF antiserum and fluorescein-conjugated goat antirabbit IgG. In both patients the majority of PMNs were DAF deficient, and these cells contained very little alkaline phosphatase activity. In contrast, the smaller, DAF-positive cell populations were phosphatase replete. This is the first demonstration that abnormalities in DAF and LAP activity occur in the same PNH PMN population and strengthens the hypothesis that defective anchoring of proteins to membrane glycophospholipid underlies the pathophysiology of this disorder.
mesh_terms: D000368:Aged; D000469:Alkaline Phosphatase / Q000097:blood / Q000172:deficiency*; D018958:CD55 Antigens; D002469:Cell Separation; D004912:Erythrocytes / Q000145:classification; D006457:Hemoglobinuria, Paroxysmal / Q000097:blood* / Q000201:enzymology; D006461:Hemolysis; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000097:blood / Q000172:deficiency*; D008875:Middle Aged; D009504:Neutrophils / Q000145:classification / Q000201:enzymology / Q000378:metabolism*
chemical_list: D018958:CD55 Antigens; D008565:Membrane Proteins; D000469:Alkaline Phosphatase