title: [Protection of PNH red blood cells from lysis by transferring GPI-anchored protein vesicles of normal human RBCs in vitro].
abstract: OBJECTIVE
  To observe the transfer of GPI-anchored protein CD59 from vesicles of normal RBCs to paroxysmal nocturnal hemoglobinuria(PNH) RBCs can correct the susceptibility to complement attack in vitro.
  
  
  METHODS
  Vesicles released from normal RBCs under ATP depletion or storage, are rich in CD59. PNH CD59- RBCs were separated by elution from immunoaffinity column bound with monoclonal antibody CD59, and then incubated with normal RBCs vesicles. The content of CD59 in vesicles and RBCs was detected by Western blot and flowcytometric analysis respectively and hemolysis of PNH CD59- RBCs was detected by cobra venom factor hemolysis test.
  
  
  RESULTS
  The fluorescence intensity of PNH CD59- RBCs were increased from (1.68 +/- 0.57)% to (58.42 +/- 8.25)% and the hemolysis was significantly decreased from (27.13 +/- 9.69)% to (19.49 +/- 7.61)% after incubation with vesicles prepared from normal donors (P < 0.05), while normal RBCs had no obvious change before and after the same treatment. (P > 0.05).
  
  
  CONCLUSIONS
  It seems that CD59 protein molecules could be transfered from normal RBCs vesicles to PNH CD59- RBCs and retaining the complement regulatory function. CD59- RBCs could be rendered less liable to hemolyze during complement attack.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: C
authors.initials: C
authors.lastname: Xu
authors.forename: H
authors.initials: H
authors.lastname: Pan
mesh_terms: D018957:CD59 Antigens / Q000378:metabolism*; D002469:Cell Separation; D004912:Erythrocytes / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006457:Hemoglobinuria, Paroxysmal / Q000378:metabolism*; D006801:Humans; D021381:Protein Transport; D022161:Transport Vesicles / Q000378:metabolism
chemical_list: D018957:CD59 Antigens; D017261:Glycosylphosphatidylinositols