Human red blood cell loading with hexokinase-inactivating antibodies. An in vitro model for enzyme deficiencies

Acta Haematologica
M MagnaniV Stocchi

Abstract

The primary cause of red cell destruction in enzymopathies of anaerobic remains controversial and difficult to investigate especially because the erythrocyte population in enzymopenic patients is largely heterogeneous. We have shown that loading human erythrocytes with monospecific enzyme-inactivating antibodies could be useful in understanding the biochemical modifications occurring in enzymopenic erythrocytes and the mechanisms leading to red cell destruction. Hexokinase-inactivating antibodies were prepared and loaded in human erythrocytes using a procedure of encapsulation based on hypotonic hemolysis, isotonic resealing and reannealing. Red blood cells loaded with anti-hexokinase IgG showed 20 +/- 3% residual hexokinase activity while all other enzymes were normal. Lactate production by these cells was 30% of controls while the amount of glucose metabolized in the hexose monophosphate pathway (HMP) was unchanged under resting conditions. However, in the presence of methylene blue HMP rates were only 12% of controls. Determination of adenine nucleotide levels suggests that the antihexokinase-loaded red blood cells are not able to maintain, in vitro, their ATP level as well as their 2,3-diphosphoglycerate. Osmotic fragility,...Continue Reading

Citations

May 26, 1993·Molecular and Cellular Biochemistry·M MagnaniG Gazzanelli
Mar 12, 2004·Journal of Controlled Release : Official Journal of the Controlled Release Society·Carmen Gutiérrez MillánJosé M Lanao
Sep 1, 1994·Cell Biochemistry and Function·L ChiarantiniM Magnani

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