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Inactivation of the mitogenic property of pokeweed mitogen by erythrocytes
Abstract:
In whole blood culture, pokeweed mitogen (PWM) is unable to stimulate human lymphocytes. This is because the mitogenic property of PWM is inactivated by erythrocytes, presumably due to absorption. The inactivation was observed with as few as 5 x 10(6)/ml of human erythrocytes. Therefore, when PWM is used to study the functions of human lymphocytes, especially of suppressor T lymphocytes, erythrocytes should be removed from lymphocyte preparations for accurate analysis.
Insights
Pokeweed mitogen (PWM) cannot stimulate human lymphocytes in whole blood cultures because red blood cells (erythrocytes) inactivate it. Removing erythrocytes is crucial for accurate analysis of lymphocyte function, particularly suppressor T cells.
Area of Science:
- Immunology
- Cell Biology
Background:
- Pokeweed mitogen (PWM) is a common stimulant used in lymphocyte research.
- Accurate analysis of human lymphocyte function requires optimal experimental conditions.
Purpose of the Study:
- To investigate the inability of pokeweed mitogen (PWM) to stimulate human lymphocytes in whole blood cultures.
- To identify the factor responsible for PWM inactivation and recommend adjustments for accurate lymphocyte analysis.
Main Methods:
- Whole blood cultures were established using human lymphocytes.
- The effect of erythrocytes (red blood cells) on PWM-induced lymphocyte stimulation was assessed.
- PWM activity was measured in the presence and absence of varying erythrocyte concentrations.
Main Results:
- Pokeweed mitogen (PWM) failed to stimulate human lymphocytes in whole blood cultures.
- The presence of human erythrocytes, even at low concentrations (5 x 10^6/ml), inactivated the mitogenic property of PWM.
- Inactivation of PWM by erythrocytes is likely due to absorption.
Conclusions:
- Erythrocytes interfere with pokeweed mitogen (PWM) stimulation of human lymphocytes.
- Lymphocyte preparations must be depleted of erythrocytes for accurate assessment of PWM-induced responses.
- This finding is particularly important for studies involving suppressor T lymphocytes.