Measurement of Lectin-Induced Superoxide Release from Human Neutrophils

  • A. V. Timoshenko
  • H.-J. Gabius
Part of the Springer Laboratory book series (SLM)


Lectin-carbohydrate interaction can trigger diverse cellular responses. Plasma membranes of human neutrophils contain NADPH oxidase (Cross and Jones 1991), which can be activated by different stimuli including lectins (Cohen et al. 1982). The key component of this superoxide anion (O 2 )- generating system, cytochrome b-245, was shown to be a glycoprotein with about 21% carbohydrate (Harper et al. 1985), even enabling direct interaction with some exogenous lectin. There are different methods to determine the activity of NADPH oxidase and respiratory burst of neutrophils: luminol-dependent chemiluminescence (Hallett and Campbell 1983), nitroblue tetrazolium dye reduction assay (Das et al. 1990), electron microscopic technique (Ohno et al. 1982), fluorometric assay (Hyslop and Sklar 1984) and reduction of cytochrome c (Markert et al. 1984). The last method is most useful due to its high specificity to O 2 . The method can be applied in various modifications: continuous and discontinuous assay (Markert et al. 1984) and microassay (Mayo and Curnutte 1990). Employing cytochrome c, any lectin-dependent effect on the generation of this compound, involved in host defence, can easily be monitored.


NADPH Oxidase Human Neutrophil Phorbol Myristate Acetate Fluorometric Assay Electron Microscopic Technique 
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© Springer-Verlag Berlin Heidelberg 1993

Authors and Affiliations

  • A. V. Timoshenko
  • H.-J. Gabius

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