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Updated: Oct 13, 2026
An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes
Published on: November 21, 2016
The effects of cAMP on the expression of glycolytic isozymes in activated peripheral human T lymphocytes
S Marjanovic1, P Wollberg, S Skog
1Department of Biochemistry, Arrhenius Laboratory, Stockholm University, Sweden.
Abstract:
Forskolin, an activator of adenylate cyclase, inhibits mitogen induction of glycolytic isozymes in human peripheral T cells. Inhibition of lactate dehydrogenase isozyme activity is correlated with lowered mRNA levels, suggesting a transcriptional block in the presence of increased cAMP. Forskolin added with the mitogen, or 12-24 h after the mitogen, strongly inhibits isozyme expression and DNA synthesis, and causes cells to accumulate in the G0 or early G1 phase of the cell cycle. The data suggest that DNA synthesis and isozyme expression are both inhibited by a cAMP-sensitive step(s) in the early activation or progression phase.
Insights
Forskolin, a cAMP activator, blocks T cell glycolytic enzyme production and DNA synthesis. This suggests a critical cAMP-dependent step regulates early T cell activation and proliferation.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- T cells play a crucial role in immune responses.
- Cellular metabolism, including glycolysis, is vital for T cell activation.
- Adenylate cyclase activators like forskolin can modulate cellular processes.
Purpose of the Study:
- To investigate the effect of forskolin on T cell activation and metabolism.
- To determine the role of cyclic AMP (cAMP) in regulating glycolytic isozyme expression during T cell activation.
Main Methods:
- Human peripheral T cells were treated with forskolin and mitogens.
- Lactate dehydrogenase (LDH) isozyme activity and mRNA levels were measured.
- Cell cycle progression and DNA synthesis were assessed.
Main Results:
- Forskolin inhibited mitogen-induced glycolytic isozyme expression in T cells.
- LDH isozyme inhibition correlated with decreased mRNA levels, indicating transcriptional regulation.
- Forskolin treatment led to cell cycle arrest in G0/early G1 phase and inhibited DNA synthesis.
Conclusions:
- Increased cAMP levels, induced by forskolin, inhibit T cell glycolytic isozyme expression and DNA synthesis.
- A cAMP-sensitive regulatory step likely controls early T cell activation and progression.
- These findings highlight the role of cAMP in modulating T cell metabolism and proliferation.
