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Updated: Jun 11, 2026
Rapid Quantification of Mitogen-induced Blastogenesis in T Lymphocytes for Identifying Immunomodulatory Drugs
Published on: December 27, 2016
Effect of mitogen lectin on lymphocyte or brain cortex cell activation
Hana Kovaru1, Frantisek Kovaru, Marek Pav
11st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic. hkova@lf1.cuni.cz
Insights
Phytohemagglutinin (PHA) activates Na+/K+-ATPase in lymphocytes and brain cells. Early enzyme activation correlates with later lymphocyte proliferation, suggesting a role in immune signaling.
Area of Science:
- Immunology
- Cell Biology
- Neuroscience
Background:
- The sodium-potassium pump (Na+/K+-ATPase) is crucial for cellular function.
- Mitogenic lectins like phytohemagglutinin (PHA) stimulate immune cells.
- Understanding early cellular responses to mitogens is key to deciphering immune activation.
Purpose of the Study:
- To investigate PHA-induced changes in Na+/K+-ATPase activity in lymphocytes and brain cells.
- To analyze the relationship between early PHA-evoked enzyme activation and subsequent lymphocyte proliferation.
Main Methods:
- Biochemical analysis of inorganic phosphate (Pi) release from ATP to measure Na+/K+-ATPase activity.
- Lymphocyte proliferation assessed via 3H-thymidine incorporation.
Main Results:
- PHA significantly stimulated Na+/K+-ATPase activity in mouse spleen lymphocytes and brain cortex cells.
- In pig lymphocytes, PHA dose and potassium ion concentration influenced Na+/K+-ATPase activity and 3H-thymidine incorporation.
- Enzyme activity stimulation was dependent on cell type and PHA dose.
Conclusions:
- Early PHA-induced Na+/K+-ATPase activation is a conserved response across species and cell types.
- The intensity of early enzyme activation may predict the level of T lymphocyte proliferation.
- Na+/K+-ATPase is implicated as a component of mitogen-activated signal transduction pathways.
Objectives:
We studied a) mitogen lectin (PHA) evoked changes of Na+/K+-ATPase activity in functionally different lymphocytes or brain cortex cells and b) quantitative relationship between PHA- evoked early enzyme activation and late lymphocyte proliferation were analyzed.
Materials And Methods:
We performed biochemical analyses of Pi released from ATP by Na+/K+-ATPase activity. Lymphocyte proliferation was assayed by 3H-thymidine incorporation.
Results:
We demonstrated PHA stimulated Na+/K+-ATPase activity of mouse spleen lymphocytes or freshly isolated brain cortex cells. Besides this, we estimated high stimulation of Na+/K+-ATPase activity and subsequent late 3H-thymidine incorporation into pig lymphocytes as both PHA dose and K+ ion concentration dependent.
Conclusions:
Thus, early PHA dose-dependent stimulation of Na+/K+-ATPase activity is a more general response in different animal species and functionally different cells. We measured both cell type- and PHA-dose dependent enzyme activity stimulation. We can suggest that intensity of early PHA induced Na+/K+-ATPase activation could be in relationship to subsequent elevated level of T lymphocyte proliferation. The Na+/K+-ATPase can be a part of mitogen lectin evoked signal transduction mechanisms.
