Interleukin 2 mRNA induction in human lymphocytes: analysis of the synergistic effect of a calcium ionophore A23187 and a phorbol ester

European Journal of Immunology - Tập 15 Số 12 - Trang 1204-1208 - 1985
Yusei Yamamoto1, Takafumi Ohmura1, Kouji Fujimoto1, Kaoru Onoue1
1Department of Biochemistry, Institute for Medical Immunology, Kumamoto University Medical School, Kumamoto

Tóm tắt

AbstractInduction of interleukin 2 (IL2) mRNA in human tonsillar lymphocytes under various conditions was examined by cytoplasmic dot hybridization using a 32P‐labeled IL 2 cDNA probe to study the signal transduction mechanisms which lead to IL2 gene expression. A tumor‐promoting phorbol ester, 12‐O‐tetradecanoyl‐phorbol 13‐ace‐tate (TPA), acted synergistically with a Ca2+ ionophore A23187 or phytohemaggluti‐nin (PHA) to induce a high level of IL2 mRNA in lymphocytes, whereas each of them by itself could not induce the mRNA production. In two‐step culture experiments the lymphocytes pulse‐incubated with TPA for 1 h (the first culture) could efficiently initiate IL2 mRNA production by subsequent culture with A23187 or PHA (the second culture). Results obtained by removal of extracellular Ca2+ from either the first or second culture revealed that Ca2+ was not necessarily required during the first culture with TPA, but it is essential in the second culture with A23187 or PHA, regardless of the presence or absence of Ca2+ in the first culture. A reagent known to be a calmodulin antagonist, N‐(6‐aminohexyl)‐5‐chloro‐l‐naphthalenesulfonamide (W‐7), almost completely inhibited the IL2 mRNA induction in A23187‐TPA‐stimu‐lated lymphocytes at a concentration of 25 μM, whereas N‐(6‐aminohexyl)‐l‐naph‐thalenesulfonamide that has much lower affinity for calmodulin than W‐7 did not inhibit at this concentration. The IL2 mRNA induction was also blocked by the addition of 50 UM of 8‐(N, N‐diethylamino)‐octyl‐3,4,5‐trimethoxybenzoate hydro‐ chloride which is known to block the release of Ca2+ from intracellular storage sites. These results show that mobilization of Ca2+ and the calmodulin‐dependent regulatory system appear to work synergistically with TPA which probably activates protein kinase C in the pathway to IL2 gene expression.

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Tài liệu tham khảo

10.1126/science.181845

10.1111/j.1600-065X.1980.tb00327.x

10.1111/j.1600-065X.1982.tb00410.x

10.1111/j.1600-065X.1982.tb00414.x

10.1111/j.1600-065X.1982.tb00415.x

10.1038/302305a0

10.1016/S0006-291X(83)80040-0

10.1073/pnas.81.8.2543

10.1073/pnas.80.24.7437

Hirano T., 1984, J. Immunol., 132, 2165, 10.4049/jimmunol.132.5.2165

10.1073/pnas.81.16.5214

10.4049/jimmunol.133.1.273

Yamamoto Y., 1985, J. Biochem., 98, 49, 10.1093/oxfordjournals.jbchem.a135271

10.1016/S0021-9258(18)34459-4

10.1038/308693a0

10.1016/S0021-9258(18)32273-7

10.4049/jimmunol.124.5.2390

10.1038/255378a0

10.1016/B978-0-12-569650-0.50012-7

10.1038/295068a0

10.4049/jimmunol.128.4.1903

10.1111/j.1348-0421.1983.tb03570.x

10.1016/S0021-9258(18)34158-9

10.1073/pnas.77.9.5201

10.1016/B978-0-12-152815-7.50006-5

10.1126/science.6243188

10.1038/285073a0

10.1016/S0076-6879(83)02019-4

10.1111/j.1476-5381.1975.tb07359.x

10.1007/BF01973834

10.1038/251324b0

10.1073/pnas.71.12.5088

10.1016/0304-4165(77)90320-8

10.1073/pnas.80.11.3444

10.1002/eji.1830140317

10.1073/pnas.81.13.4169

Farrar J. J., 1980, J. Immunol., 725, 793, 10.4049/jimmunol.125.2.793

Koretzky G. A., 1982, J. Immunol., 128, 1776, 10.4049/jimmunol.128.4.1776

10.1016/0008-8749(81)90157-X

Tanaka T., 1982, Mol. Pharmacol., 22, 408

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European Journal of Immunology

Tập 15 Số 12

1204-1208

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