Immunological and physiological characteristics of the rapid immune hemolysis of neuraminidase-treated sheep red cells produced by fresh guinea pig serum

The rapid hemolysis by fresh guinea pig serum known to occur with neuraminidase-treated sheep red cells has been investigated with respect to the immunological and physiological properties of the lytic process. The following observations were made: (a) The susceptibility to hemolysis is directly proportional to the amounts of neuraminic acid enzymatically released from the cell surface. Complement lysis is mediated through binding of an IgM antibody protein to membranes of neuraminidase-treated cells. (b) Hemolysis is relatively temperature-independent above about 28 degrees C but below which a decrease in the hemolysis rate occurs. Arrhenius activation energies above and below the transition temperature were therefore found to be different. (c) Colloid osmotic swelling of neuraminidase-treated high potassium sheep red cells with a chloride ion concentration ratio near unity suspended in high potassium medium could not be prevented by sucrose. Hence, colloid osmotic swelling before lysis must be due to the entrance of sucrose and water since sucrose was the only external solute not at equilibrium. (d) From the rate of swelling and the apparent flux of sucrose under nonsteady state conditions an experimental permeability coefficient (P) for sucrose of 3-10(-8) cm-s-1 was computed. Comparison with a theoretical P of 4-10(-6) cm-s-1 for sucrose freely permeating through a single, hypothetical membrane lesion per cell of 60 A effective diameter indicates a membrane lesion which permits the passage of solutes larger than cations, but clearly constrains the free diffusion of sucrose.