Adenosine Triphosphate Metabolism in Hereditary Spherocytosis*

Abstract

The red cell in hereditary spherocytosis (HS) is unusually susceptible to the damaging effects of erythrostasis. The spleen is the primary site of erythrostasis in vivo, and it is a feature of HS that splenectomy corrects the hemolytic anemia, although the intrinsic red cell defect persists. The characteristic laboratory findings in HS of an increase in osmotic fragility and in autohemolysis are enhanced by in vitro incubation that simulates in vivo erythrostasis. Since these in vitro abnormalities are partially corrected by the addition of glucose (2), efforts have been made to detect a defect in the carbohydrate metabolism of the HS erythrocyte. As yet no definite abnormality in glycolysis has been demonstrated (3-5).In 1953 Harris and Prankerd (6) showed that there was an increased rate of extrusion of Na by the HS red cell, and in 1957 Bertles (7) demonstrated an increased rate of influx of Na into the HS erythrocyte. Jacob and Jandl (8) have confirmed this earlier work and have observed an increased turnover of Na by the HS red cell that is approximately twice normal. Based on these observations, they have postulated that the primary defect in HS is an abnormal permeability of the red cell to the passive influx of Na with secondary changes in glycolysis. To maintain a high concentration of K and a low concentration of Na within the red cell with respect to the concentration of these cations in the plasma, active transport against a gradient must take place. In the HS * Submitted for publication December 15, 1964; accepted May 10, 1965. This investigation was supported by grant HE-06239 from the National Heart Institute.Part of this work was presented at the national meeting of the American Federation for Clinical Research, Atlantic City, N. J., May 3, 1964 (1). red cell there is an increased passive influx of Na (7, 8) that must be compensated by increased active efflux of Na if osmotic hemolysis is to be prevented. Jacob and Jandl (8) have demonstrated that maneuvers which interfere with this compensatory increase in active Na transport lead to an increased rate of destruction of HS red cells in vivo. Since the energy necessary for the active transport of Na is derived from glycolysis in the form of adenosine triphosphate (ATP) (9-12), increased utilization of ATP and glucose by the HS red cell would be anticipated. Furthermore, ouabain, which blocks the utilization of ATP by the Na-K pump (11-13), should nullify increased rates of ATP and glucose utilization by HS red cells if the increase is caused by hyperactivity of cation transport. The data presented in this paper indicate an increased rate of ATP and glucose utilization by the HS erythrocyte that is only partially abolished by ouabain. The results suggest that in addition to increased utilization of ATP for active cation transport, the HS erythrocyte has an increased requirement of ATP for other metabolic processes as well. MethodsSubjects studied. HS red cells were obtained from eight patients. These patients were from three families, and all but...

Metabolic dependence of the critical hemolytic volume of human erythrocytes: relationship to osmotic fragility and autohemolysis in hereditary spherocytosis and normal red cells.

“…Although the addition of ouabain appears to increase slightly the 48-hour hemolysis of HS blood with glucose, this effect at both 24 and 48 hours is minimal compared with the significant protection against hemolysis conferred by glucose even in the presence of ouabain. The latter finding is at variance with the findings of Mohler (25), who demonstrated no protection by glucose in the presence of ouabain. Two of Mohler's experimental conditions differed from ours, continuous mixing of the blood and a lower concentration of ouabain (2.5 X 10-5 mole per L).…”

“…Mohler (25) presented evidence that ATP utilization by HS cells is greater than normal even in the presence of ouabain; therefore, these levels were compared after 24 hours of incubation. Table II significant (p < 0.01), whereas the difference between their total cation content is significant at the p < 0.05 level.…”

The role of membrane lipids in the survival of red cells in hereditary spherocytosis

“…As has been observed in normal red cells (12), this gain in cholesterol in vitro was not associated with (4,35). This leads to an increased rate of sodium extrusion and an accelerated rate of gucose consumption (4,36). To test whether the beneficial effect of obstructive jaundice on HS cells in vivo was related to a change in osmotic fragility alone, or to an alteration in cell membrane permeability and glucose metabolism as well, red cells from two patients with HS who had undergone splenectomy and from two normal subjects were incubated in media containing normal heated serum or heated serum from a patient with obstructive jaundice.…”

A New Familial Disorder with Abnormal Erythrocyte Morphology and Increased Permeability of the Erythrocytes to Sodium and Potassium

“…A further point of similarity is the apparent capacity for essentially complete compensation in vitro for the cation permeability defect. It has been demonstrated that erythrocytes in hereditary spherocytosis are able to compensate for a defect in sodium permeability by augmented sodium transport, which is accompanied by an increased rate of glycolysis [I13 and adenosine triphosphate production [16]. In the disorder presented in this report the increased rate of glycolysis by the erythrocytes and the accelerated efflux of intracellular sodium under glycolytic conditions probably reflect a similar compensatory process.…”