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Removal of contaminating polymorphonuclear leukocytes to prepare purified lymphocytes for cytotoxicity testing
Abstract:
Human polymorphonuclear leukocytes (PMN) die, apparently nonspecifically, in the standard tissue typing assay. When the cell preparation contains more than 20% PMN, proper evaluation of the histocompatibility assay is difficult. PMN were noted to aggregate when normal rabbit serum was added as the complement source. Based on the latter observation, a simple technique was developed which reduces the PMN contamination in the suspension of test cells and returns high yields of lymphocytes. Removal of the PMN lowers the high background of dead cells and permits precise evaluation of the microcytotoxicity test results.
Insights
Human polymorphonuclear leukocytes (PMN) can interfere with histocompatibility testing. A new method effectively removes PMN, improving the accuracy of microcytotoxicity test results by reducing dead cell background.
Area of Science:
- Immunology
- Cell Biology
- Histocompatibility Testing
Background:
- Human polymorphonuclear leukocytes (PMN) exhibit non-specific cell death during standard tissue typing assays.
- High concentrations of PMN (>20%) in cell preparations complicate the accurate evaluation of histocompatibility assays.
- PMN aggregation observed with normal rabbit serum (complement source) provided a basis for developing a new technique.
Purpose of the Study:
- To develop a simple and effective method for reducing polymorphonuclear leukocyte (PMN) contamination in lymphocyte preparations.
- To improve the accuracy and reliability of histocompatibility testing by minimizing PMN-related interference.
- To enhance the evaluation of microcytotoxicity test results.
Main Methods:
- A novel technique was developed leveraging PMN aggregation in the presence of normal rabbit serum.
- This method selectively removes PMN from lymphocyte suspensions.
- The procedure aims to maintain high yields of viable lymphocytes.
Main Results:
- The developed technique successfully reduces PMN contamination in cell suspensions.
- The removal of PMN significantly lowers the background of dead cells in the assay.
- This leads to a precise and reliable evaluation of microcytotoxicity test results.
Conclusions:
- A simple method for PMN removal effectively enhances histocompatibility assay accuracy.
- Reducing PMN contamination is crucial for precise microcytotoxicity testing.
- This technique offers a valuable improvement for immunological and transplantation diagnostics.
