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A novel cell surface antigen, 4C8, is expressed on human eosinophils
Yasutaka Aoyama1, Takahisa Yamane, Masayuki Hino
1Department of Clinical Hematology, Osaka City University Medical School, Osaka, Japan.
Insights
A novel monoclonal antibody, anti-4C8, identifies a new cell surface antigen on human peripheral eosinophils. This discovery provides a valuable marker for distinguishing eosinophils from neutrophils in research and clinical settings.
Area of Science:
- Immunology
- Hematology
- Cell Biology
Background:
- A novel monoclonal antibody, anti-4C8, recognizes human peripheral lymphocytes and monocytes.
- The 4C8 antigen is not expressed on neutrophils.
- The expression of 4C8 on human peripheral eosinophils was previously uncharacterized.
Purpose of the Study:
- To investigate the expression of the 4C8 antigen on human peripheral eosinophils.
- To determine if 4C8 can serve as a specific marker for eosinophils.
Main Methods:
- Flow cytometry was used to analyze 4C8 antigen expression on eosinophils.
- Western blotting was employed for molecular analysis of the 4C8 antigen in eosinophils.
- Eosinophils were purified from peripheral blood for detailed analysis.
Main Results:
- The 4C8 antigen is expressed on CD16-negative granulocytes, which include eosinophils.
- Flow cytometry confirmed anti-4C8 antibody reactivity with purified human eosinophils.
- Western blotting detected a single 80 kDa band for the 4C8 antigen in eosinophil lysates.
- A strong correlation (r = 0.91, P < 0.0001) was observed between eosinophil percentage and 4C8-positive/CD16-negative cells.
Conclusions:
- The novel 4C8 cell surface antigen is demonstrably expressed on human peripheral eosinophils.
- 4C8 represents a valuable new marker for identifying human eosinophils, aiding in their differentiation from neutrophils.
Background:
A novel monoclonal antibody, anti-4C8, reacted with human peripheral lymphocytes and monocytes but not with neutrophils. In this study, we investigated whether the 4C8 antigen is expressed on human peripheral eosinophils.
Methods:
Expression of the 4C8 antigen on eosinophils was analyzed by flow cytometry and molecular analysis of the antigen was performed with eosinophils by Western blotting.
Results:
Among human peripheral granulocytes, the 4C8 antigen was expressed on CD16-negative cells but not on CD16-positive cells. The 4C8 antigen also appeared to be expressed on eosinophils. To confirm the latter finding, eosinophils were purified from peripheral blood. On flow cytometric analysis, anti-4C8 antibody reacted with purified eosinophils. On Western blotting analysis, anti-4C8 reacted with a single band of 80 kDa in lysates from purified eosinophils. The correlation between the percentage of eosinophils determined by May-Giemsa staining and the percentage of 4C8-positive/CD16-negative cells among granulocytes was good (r = 0.91, P < 0.0001).
Conclusions:
Only a few cell surface antigens are available to distinguish human peripheral eosinophils from neutrophils. The novel cell surface antigen, 4C8, is a useful new marker of human eosinophils.
