Increase in band 3 density and aggregation in hereditary spherocytosis.

Purpose: Red cells in hereditary spherocytosis are characterized by a reduced surface area/volume ratio. The mechanisms leading to the loss of membrane material and subsequent elimination of the cells have still not been clarified. It was the aim of the present study to analyze band 3 distribution in the red cell membrane and its putative role in red cell elimination.

Methods/results: Immunogold histochemistry was performed to detect band 3 in red cell membranes. Band 3 density and distribution were visualized by electron microscopy. Unsplenectomized spherocytosis patients (n = 12) showed reduced band 3 density and aggregation compared to controls (n = 15) (density: 1.2 +/- 0.1 gold particles/microm circumference of red cell membrane vs 1.5 +/- 0.07 gold particles/microm, x +/- SEM; P < 0.05; aggregation: 0.26 +/- 0.02 aggregates/microm vs 0.3 +/- 0.02 aggregates/microm). By contrast, band 3 density and aggregation were increased in spherocytosis patients who had undergone splenectomy (density: 2.8 +/- 0.1 gold particles/microm vs 2.0 +/- 0.1 gold particles/microm; P < 0.05; aggregation: 1.5 +/- 0.1 aggregates/microm vs 0.5 +/- 0.03 aggregates/microm; P < 0.01). Artificial ageing of red cells from healthy controls (n = 6) led to a significant increase in band 3 aggregation (2.06 +/- 0.2 aggregates/microm vs 0.33 +/- 0.1 aggregates/microm; P(Wilcoxon) < 0.01) but no change in band 3 density. In hereditary spherocytosis (n = 6), both band 3 density and aggregation increased significantly after artificial ageing of the red cells. The elevated band 3 aggregation was associated with a stimulated erythrophagocytosis in vitro.

Conclusion: Band 3 aggregation characterizes the red cells in hereditary spherocytosis. It may be the cause of selective splenic phagocytosis of both spherocytes and senescent erythrocytes.