Effect of the In Vitro Ageing on the Restriction of Sheep Erythrocyte Lysis by Horse Serum

Abstract

Fresh sheep erythrocytes are resistant to lysis by horse serum but gradually became susceptible to this source of complement after storage (ageing) in the blood at 4 degrees C. This occurs faster when the fresh cells are stored in isotonic buffer. The supernatant of the buffer/red cell suspension stored at 4 degrees C for 10-15 days restrict lysis by horse serum when incubated back at 37 degrees C or 43 degrees C with the 'aged' sheep cells or fresh guinea pig red cells. In assays using fresh guinea pig erythrocytes this effect is described by reincubation of the cells in buffer, and is specific to horse serum when compared to human serum.

Determination of CD59 protein in normal human serum by enzyme immunoassay, using octyl-glucoside detergent to release glycosyl-phosphatidylinositol-CD59 from lipid complex

“…Constitutive levels of CD59 originating from several types of cells and detected in biological fluids, may be an additional physiological mechanism of cellular protection against complement-mediated lysis in infection or inflammation [12]. Complement membrane regulators may be shed by erythrocytes and be reinserted into both homologous and heterologous cells, thus restricting lysis [21,22]. In vitro and in vivo studies have shown that GPI-anchored proteins can be transferred between cells in microvesicles or in high-density lipoprotein (HDL) particles [19], and it has been suggested that this mechanism could be of help in therapeutic processes such as the transfer of CD59 to the red cells of patients with paroxysmal nocturnal hemoglobinuria (PNH) [23].…”