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Published on: September 10, 2019
Lectinocytochemical specificity in human eosinophils and neutrophils: a reexamination
1Cryobiology Research Center, University of Texas Health Science Center, Houston 77030.
Insights
This study reveals lectin binding patterns in human leukocyte granules using advanced cryofixation techniques. Soybean agglutinin and wheat germ agglutinin bind to specific granule matrices in eosinophils and neutrophils.
Area of Science:
- Cell Biology
- Immunology
- Biochemistry
Background:
- Leukocyte cytoplasmic granules contain glycoproteins, which are targets for lectin binding.
- Traditional wet chemical processing can alter granule composition, affecting lectin localization studies.
- Specific lectin-cytochemical protocols are crucial for accurate analysis of granule components.
Purpose of the Study:
- To investigate the localization of specific lectins within human eosinophil and neutrophil granules.
- To evaluate the impact of artifact-free specimen preparation on lectin-gold electron microscopy.
- To identify carbohydrate moieties in leukocyte granule matrices using lectin binding.
Main Methods:
- Isolated human eosinophils and neutrophils were prepared using rapid cryofixation and molecular distillation drying.
- Lectin-gold electron microscopy was employed for localization studies.
- Soybean agglutinin (SBA) and wheat germ agglutinin (WGA) were used as specific lectins.
Main Results:
- Soybean agglutinin (SBA) specifically bound to eosinophil granule matrices and a subset of neutrophil granules.
- Wheat germ agglutinin (WGA) showed a similar labeling pattern, confirming the presence of N-acetyl-D-galactosamine and N-acetyl-D-glucosamine residues.
- These findings contrast with previous reports regarding lectin binding to crystalline cores.
Conclusions:
- Advanced cryofixation methods preserve leukocyte granule integrity for accurate lectin localization.
- Specific lectin binding patterns highlight the presence of distinct carbohydrate structures in granule matrices.
- Careful specimen preparation is essential for reliable lectinocytochemical studies.
Abstract:
As with secretory granules in other cell types, many of the protein components that make up the cytoplasmic granules of human leukocytes are glycoproteins. It is not unexpected, therefore, that lectins specific for various carbohydrate moieties can be localized in these granules following appropriate protocols for specimen preparation and thin-section labeling. In this study, isolated human eosinophils and neutrophils were prepared for lectin-gold electron microscopic localization by procedures that involve no exposure to aqueous fixatives, buffers, or solvents (rapid cryofixation and molecular distillation drying), thus removing the potential problem of constituent extraction or translocation during so-called "wet chemical" processing. In contrast to other reports, data presented illustrate the specific binding of soybean agglutinin (SBA) to eosinophil granule matrices (not the crystalline cores), as well as to a population of granules in neutrophils. A similar labeling pattern for wheat germ agglutinin (WGA) was also seen, confirming the presence of N-acetyl-D-galactosamine and N-acetyl-D-glucosamine residues in eosinophil and neutrophil granule matrices. These studies emphasize the need for carefully designed specimen preparation as well as subsequent thin-section labeling procedures in lectinocytochemical studies.
