Stimulatory Roles of Muscarinic Acetylcholine Receptors on T Cell Antigen Receptor/CD3 Complex-Mediated Interleukin-2 Production in Human Peripheral Blood Lymphocytes
- 1Department of Pharmacology, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060, Japan (H.F., Y.K., T.Y., T.U., Y.N.), and 2Department of Neuroscience, Research Institute for Oriental Medicine, Toyama Medical and Pharmaceutical University, Toyama 930–01, Japan (Y.N.)
Abstract
It is known that there are some bidirectional interactions between the nervous and the immune systems via neurotransmitters and cytokines. To clarify whether any neurotransmitters modulate lymphocyte functions, we examined the effects of oxotremorine-M (Oxo-M) on interleukin-2 (IL-2) production in human peripheral blood lymphocytes by using enzyme-linked immunosorbent assays, Northern blot analyses, reverse transcriptase-polymerase chain reaction, and fluorescence-activated cell sorter. Pretreatment of cells with Oxo-M (10 nm to 10 μm) for 4–24 hr enhanced phytohemagglutinin (PHA)-induced IL-2 mRNA expression and markedly increased IL-2 production compared with those induced by PHA alone. Oxo-M alone did not affect IL-2 mRNA expression and IL-2 production. In CD3-positive T cells, pretreatment with Oxo-M for 24 hr enhanced PHA-induced IL-2 production. Furthermore, pretreatment with Oxo-M enhanced PHA-induced mRNA expression of the α and β subunits of IL-2 receptors and DNA synthesis. Cytometric analysis showed Oxo-M treatment did not up-regulate expression of cell surface molecules such as CD3, CD2, CD4, CD8, and IL-2 receptors. These results suggest that activation of muscarinic receptors enhances T cell antigen receptor/CD3-induced IL-2 production.
Footnotes
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Send reprint requests to: Yasuyuki Nomura, Ph.D., Department of Pharmacology, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060, Japan. E-mail:nomura{at}pharm.hokudai.ac.jp
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This work was supported by Special Coordination Funds for Promoting Science and Technology from the Science and Technology Agency and by a Grant-in-Aid from the Ministry of Education, Science and Culture in Japan.
- Abbreviations:
- ACh
- acetylcholine
- TCR
- T cell antigen receptor
- mAb
- monoclonal antibody
- PLC
- phospholipase C
- PKC
- protein kinase C
- IL-2
- interleukin-2
- CsA
- cyclosporin A
- Oxo-M
- oxotremorine-M
- PHA
- phytohemagglutinin
- TPA
- 12-O-tetradecanoylphorbol-13-acetate
- DiOC6
- 3,3′-dihexyloxacarbocyanine iodide
- TdR
- [methyl-3H]thymidine (thymine deoxyriboside)
- hPBL
- human peripheral blood lymphocytes
- ELISA
- enzyme-linked immunosorbent assay
- FACS
- fluorescence-activated cell sorter
- RT
- reverse transcription
- PCR
- polymerase chain reaction
- [Ca2+]i
- intracellular Ca2+concentration
- FCS
- fetal calf serum
- SSC
- standard saline citrate
- HEPES
- 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- AM
- acetoxymethyl ester
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- Received July 26, 1996.
- Accepted March 11, 1997.
- The American Society for Pharmacology and Experimental Therapeutics
