Effect of Metabolic State on Immune-Hemolysis of L-Positive Low Potassium (LK) Sheep Red Blood Cells by ISO-Immune Anti-L Serum and Rabbit Serum Complement

Abstract

L-antigen-positive low potassium type (LK) sheep red blood cells (SRBC) metabolically depleted by (A) starvation in glucose-free media, (B) incubation in 2-deoxy-D-glucose, or (C) exposure to iodoacetamide (IAA) were more susceptible to immune-hemolysis by ovine anti-L serum and rabbit serum complement (RSC) than control LK SRBC. The magnitude of this effect depended on the concentrations of L-antiserum and RSC used. Although all three treatments leading to enhanced immune-hemolysis of LL or LM (LK) SRBC also resulted in a loss of intracellular adenosine triphosphate [ATPl_i, the data do not permit conclusion of a direct correlation between immune-hemolysis and [ATP]_i in these cells. Restoration of [ATP]_i in glucose-containing media of LK SRBC previously metabolically depleted by treatments (A) or (B) was complete within two hours; it was followed, however, by a much slower and only partial return of immune-hemolysis to control values. The depletion induced changes appear to promote binding of RSC to, and/or enhance its action on LK SRBC sensitized with the IgG-type L-antibody rather than to unmask hidden L-antigenic sites. Thus the metabolic state becomes an important determinant of the RSC-mediated immune-hemolysis of L-antiserum exposed LL or LM (LK) SRBC.