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A comparison of three reverse haemolytic plaque assays
Immunobiology
|May 1, 1984
Summary
Reverse hemolytic plaque assays quantified plaque-forming cells (PFC) in mononuclear cells (MNC). The fluid phase assay proved most sensitive, detecting higher PFC levels in pokeweed mitogen (PWM) stimulated cultures compared to solid phase methods.
Area of Science:
- Immunology
- Cell Biology
Background:
- Quantifying antibody-secreting cells is crucial for understanding immune responses.
- Reverse hemolytic plaque (RHP) assays are established methods for this purpose.
- Comparing different RHP assay formats can optimize sensitivity and accuracy.
Purpose of the Study:
- To compare the sensitivity and correlation of fluid phase and solid phase RHP assays.
- To quantitate plaque-forming cells (PFC) in pokeweed mitogen (PWM) stimulated and unstimulated mononuclear cells (MNC).
- To assess the utility of these assays in identifying donors with varying immune responses.
Main Methods:
- Utilized one fluid phase and two solid phase RHP assays.
- Assayed mononuclear cells (MNC) from 18 normal donors.
- Included both pokeweed mitogen (PWM) stimulated and unstimulated cell cultures.
Main Results:
- A close correlation was observed between the three RHP assays.
- Assay results strongly correlated with the number of cells containing cytoplasmic immunoglobulin (cIg).
- The fluid phase RHP assay demonstrated 2-4 times higher PFC values in PWM-stimulated cultures, indicating superior sensitivity.
Conclusions:
- All three RHP assays effectively identified donors with low PWM responses.
- The fluid phase RHP assay is the most sensitive method for quantifying PFC.
- This finding has implications for accurate assessment of B-cell function and immune status.