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Updated: May 22, 2026

Qualitative and Quantitative Analysis of the Immune Synapse in the Human System Using Imaging Flow Cytometry
Published on: January 7, 2019
Regulatory mechanisms of acetylcholine synthesis and release by T cells
Takeshi Fujii1, Yuki Takada-Takatori, Koichiro Kawashima
1Department of Pharmacology, Faculty of Pharmaceutical Sciences, Doshisha Women's College of Liberal Arts, Kyotanabe, Kyoto 610-0395, Japan. tfujii@dwc.doshisha.ac.jp
Aims:
Muscarinic and nicotinic acetylcholine (ACh) receptors are expressed in immune cells. ACh synthesized by choline acetyltransferase (ChAT) and released in T cells binds to these receptors. Furthermore, we have recently demonstrated the involvement of mediatophore, a homooligomer of a 16-kDa proteolipid subunit of vacuolar H(+)-ATPase, in ACh release from T cells. In this study, we investigated the effects of phorbol 12-myristate 13-acetate (PMA), dibutyryl cAMP (dbcAMP) and FK506, an immunosuppressant calcineurin inhibitor, on lymphocytic cholinergic activity in T cells.
Main Methods:
We determined the content and release of ACh in human leukemic T cell line MOLT-3 cells using a sensitive and specific radioimmunoassay for ACh. In addition, expression of ChAT mRNA and ChAT activity were investigated using reverse-transcription-polymerase chain reaction and Fonnum method, respectively.
Key Findings:
Phytohemagglutinin (PHA), a T-cell activator, up-regulated ChAT mRNA expression, synthesis and release of ACh. PMA, a protein kinase C (PKC) activator, and dbcAMP, a protein kinase A (PKA) activator, also increased ChAT activity and ACh synthesis by up-regulating ChAT gene expression. FK506 inhibited PHA-induced up-regulation of ChAT mRNA expression, suggesting the involvement of calcineurin-mediated pathways in ChAT gene transcription.
Significance:
Activation of PKC and PKA up-regulates ACh synthesis in T cells, and immunological activation triggers ChAT gene transcription through calcineurin-mediated pathways.
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