Transfer of Prostasomal CD59 to CD59-Deficient Red Blood Cells Results in Protection against Complement-Mediated Hemolysis

Authors


Address correspondence to Bo Nilsson, Department of Clinical Immunology, Uppsala University Hospital, 751 85 Uppsala, Sweden. E-mail: bo.nilsson@klinimm.uu.se

Abstract

PROBLEM: Prostasomes isolated from human seminal plasma have complement regulatory properties because of their content of CD59, a glycosylphosphatidylinositol (GPI)-anchored protein. We investigated a functional role of prostasomes by the possibility of transferring CD59 from prostasomes to rabbit erythrocytes (RE) and human erythrocytes obtained from patients with paroxysmal nocturnal hemoglobinuria (PNH), both types of cells lacking CD59.

METHOD OF STUDY: We used the assay of hemolytic activity of the alternative pathway of the complement system to compare the liability of the erythrocytes to hemolysis by the complement system with and without pre-incubation with prostasomes. CD59 gained by the RE and PNH erythrocytes was established by flow cytometry. The effect of phosphatidylinositol phospholipase C (PIPLC) on the GPI anchor of prostasomal CD59 and the effect of heat treatment on the prostasomes were also studied. Anti-CD59 antibodies were used to block the protective effect of prostasomes on erythrocytes.

RESULTS: Both RE and PNH erythrocytes showed diminished complement-mediated hemolysis after incubation with prostasomes. This was because of the transfer of CD59 from prostasomes to the red blood cells during pre-incubation as evidenced by the hemolytic assay and flow-cytometry. The efficacy of the prostasomes was affected by heat treatment and was totally lost at 100°C. Phosphatidylinositol phospholipase C broke the GPI anchor and released CD59 from prostasomes and the RE surface (after pre-incubation with prostasomes) but not from the human PNH erythrocytes.

CONCLUSIONS: A transfer mechanism of CD59 takes place during pre-incubation from prostasomes to erythrocytes lacking CD59 which supports the idea that transfer of prostasomal CD59 can protect cells from lysis elicited by C5b-9. This might be a mechanism by which autologous and allogeneic cells are protected against complement attack in the genital tracts.

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