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IL-1 beta modulates the concanavalin-A-induced expression of proenkephalin A mRNA in murine thymocytes
K M Linner1, S E Nicol, B M Sharp
1Endocrine-Neuroscience Research Laboratory, Minneapolis Medical Research Foundation, Minnesota.
Abstract:
We have previously shown that proenkephalin A (PEA) messenger RNA (mRNA) is induced in murine thymocytes by the T cell-specific mitogen concanavalin-A (Con-A). We now show that this Con-A-induced expression of PEA mRNA is modulated by the cytokine murine interleukin-1 beta (mIL-1 beta) in a biphasic, dose-dependent manner. Murine thymocytes were cultured for 72 h with Con-A and with varying concentrations of mIL-1 beta. PEA mRNA expression was analyzed by Northern gel and solution hybridization techniques. Concentrations of mIL-1 beta of 10(-14) and 10(-13) M enhanced the Con-A-induced expression of PEA mRNA in cultured murine thymocytes up to 2.5-fold, whereas higher concentrations of mIL-1 beta (10(-11) and 10(-10) M) inhibited its expression 60 and 85%, respectively. Both the enhancing and inhibiting effects of mIL-1 beta in the Con-A-induced expression of PEA mRNA were reversed by a 100-fold excess of interleukin-1 receptor antagonist protein, but not by a 10-fold excess of interleukin-1 receptor antagonist protein. The effects of mIL-1 beta on PEA mRNA expression in Con-A-activated thymocytes are different from its effects on Con-A-stimulated thymocyte proliferation. In the latter case, only enhancement of thymocyte proliferation was seen, as measured by [3H]thymidine incorporation. The present study demonstrates that PEA mRNA expression is regulated by IL-1 beta, which is thought to play a role in thymocyte maturation.
Insights
Murine interleukin-1 beta (mIL-1 beta) modulates proenkephalin A (PEA) mRNA expression in thymocytes. Low mIL-1 beta concentrations enhance PEA mRNA, while high concentrations inhibit it, impacting thymocyte maturation.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Proenkephalin A (PEA) messenger RNA (mRNA) is induced in murine thymocytes by concanavalin-A (Con-A).
- Interleukin-1 beta (IL-1 beta) is a cytokine involved in immune responses and T cell development.
Purpose of the Study:
- To investigate the modulatory effects of murine interleukin-1 beta (mIL-1 beta) on Con-A-induced PEA mRNA expression in murine thymocytes.
- To compare the effects of mIL-1 beta on PEA mRNA expression versus thymocyte proliferation.
Main Methods:
- Murine thymocytes were cultured with Con-A and varying concentrations of mIL-1 beta for 72 hours.
- PEA mRNA expression was quantified using Northern gel and solution hybridization techniques.
- The role of interleukin-1 receptor antagonist protein was assessed to confirm specificity.
Main Results:
- Low concentrations of mIL-1 beta (10^-14 to 10^-13 M) enhanced Con-A-induced PEA mRNA expression up to 2.5-fold.
- Higher concentrations of mIL-1 beta (10^-11 to 10^-10 M) inhibited PEA mRNA expression by 60-85%.
- Both enhancement and inhibition were reversed by a 100-fold excess of interleukin-1 receptor antagonist protein, indicating IL-1 beta specificity.
Conclusions:
- Murine interleukin-1 beta (mIL-1 beta) regulates proenkephalin A (PEA) mRNA expression in a biphasic, dose-dependent manner in activated thymocytes.
- These findings suggest a role for IL-1 beta in thymocyte maturation through modulation of PEA mRNA expression.
- The effects of IL-1 beta on PEA mRNA expression differ from its effects on thymocyte proliferation, highlighting distinct regulatory pathways.
