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A general method for enumerating single cells secreting antigen: albumin-secreting hepatocytes detected as
Abstract:
Sheep erythrocytes, artificially coated with purified antibody specific for mouse serum albumin (anti-MSA Ab-E) were agglutinated and lysed by mouse serum albumin. Mouse liver cells, plated with anti-MSA Ab-E formed hemolytic plaque in a modified plaque-forming cell (PFC) assay. The same cells tested with anti-MSA Ab-E did not rosette in modified immunocyto-adhesion assay. Thus, liver cells secreted molecules (albumin) which were not detectable on the cell membrane. This PFC assay with Ab-E is probably a general method for enumerating antigen-secreting cell.
Insights
Mouse liver cells secrete albumin, a molecule not found on their surface. A modified plaque-forming cell (PFC) assay using antibody-coated erythrocytes can detect these antigen-secreting cells.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Antibodies are crucial for immune responses and cellular interactions.
- Detecting secreted antigens from cells is vital for understanding cellular function and disease.
- Current methods may not effectively identify cells secreting molecules not present on their surface.
Purpose of the Study:
- To investigate whether mouse liver cells secrete albumin.
- To develop a method for detecting antigen-secreting cells.
- To differentiate between cell-surface antigens and secreted molecules.
Main Methods:
- Sheep erythrocytes were coated with anti-mouse serum albumin antibody (anti-MSA Ab-E).
- Mouse liver cells were analyzed using a modified plaque-forming cell (PFC) assay with anti-MSA Ab-E.
- A modified immunocyto-adhesion assay was used for comparison.
Main Results:
- Mouse serum albumin caused agglutination and lysis of antibody-coated erythrocytes.
- Mouse liver cells formed hemolytic plaques in the modified PFC assay, indicating albumin secretion.
- The same liver cells did not form rosettes in the immunocyto-adhesion assay, suggesting albumin is secreted, not cell-surface bound.
Conclusions:
- Mouse liver cells secrete albumin, which is not detectable on the cell membrane.
- The modified PFC assay using antibody-coated erythrocytes is a viable method for enumerating antigen-secreting cells.
- This assay may serve as a general technique for identifying cells that secrete specific antigens.