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Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent inhibition of IL-5 from human T lymphocytes is not mediated by the cAMP-dependent protein kinase A

Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent inhibition of IL-5 from human T lymphocytes is not mediated by the cAMP-dependent protein kinase A
Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent inhibition of IL-5 from human T lymphocytes is not mediated by the cAMP-dependent protein kinase A
IL-5 is implicated in the pathogenesis of asthma and is predominantly released from T lymphocytes of the Th2 phenotype. In anti-CD3 plus anti-CD28-stimulated PBMC, albuterol, isoproterenol, rolipram, PGE2, forskolin, cholera toxin, and the cAMP analog, 8-bromoadenosine cAMP (8-Br-cAMP) all inhibited the release of IL-5 and lymphocyte proliferation. Although all of the above compounds share the ability to increase intracellular cAMP levels and activate protein kinase (PK) A, the PKA inhibitor H-89 failed to ablate the inhibition of IL-5 production mediated by 8-Br-cAMP, rolipram, forskolin, or PGE2. Similarly, H-89 had no effect on the cAMP-mediated inhibition of lymphocyte proliferation. Significantly, these observations occurred at a concentration of H-89 (3 ?M) that inhibited both PKA activity and CREB phosphorylation in intact cells. Additional studies showed that the PKA inhibitors H-8, 8-(4-chlorophenylthio) adenosine-3?,5?-cyclic monophosphorothioate Rp isomer, and a myristolated PKA inhibitor peptide also failed to block the 8-Br-cAMP-mediated inhibition of IL-5 release from PBMC. Likewise, a role for PKG was considered unlikely because both activators and inhibitors of this enzyme had no effect on IL-5 release. Western blotting identified Rap1, a downstream target of the cAMP-binding proteins, exchange protein directly activated by cAMP/cAMP-guanine nucleotide exchange factors 1 and 2, in PBMC. However, Rap1 activation assays revealed that this pathway is also unlikely to be involved in the cAMP-mediated inhibition of IL-5. Taken together, these results indicate that cAMP-elevating agents inhibit IL-5 release from PBMC by a novel cAMP-dependent mechanism that does not involve the activation of PKA
0022-1767
2074-2080
Staples, K.J.
e0e9d80f-0aed-435f-bd75-0c8818491fee
Bergmann, M.
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Tomita, K.
1924da17-4342-4997-bbc0-52d3cfc2ae15
Houslay, M.D.
ad664bab-f4e3-406f-a2ec-18609b9c5236
McPhee, I.
07f86866-ad5d-4463-8383-dbd84e03b57c
Barnes, P.J.
c20c99d4-6dd6-4975-bc1a-cf3ec17f3cee
Giembycz, M.A.
db4c128b-7598-4262-9e57-ccb8d66bc074
Newton, R.
5ec87291-b3f9-4d33-9020-d9be787919e2
Staples, K.J.
e0e9d80f-0aed-435f-bd75-0c8818491fee
Bergmann, M.
8f6c4919-eada-45d2-8548-9bfb353b5d0c
Tomita, K.
1924da17-4342-4997-bbc0-52d3cfc2ae15
Houslay, M.D.
ad664bab-f4e3-406f-a2ec-18609b9c5236
McPhee, I.
07f86866-ad5d-4463-8383-dbd84e03b57c
Barnes, P.J.
c20c99d4-6dd6-4975-bc1a-cf3ec17f3cee
Giembycz, M.A.
db4c128b-7598-4262-9e57-ccb8d66bc074
Newton, R.
5ec87291-b3f9-4d33-9020-d9be787919e2

Staples, K.J., Bergmann, M., Tomita, K., Houslay, M.D., McPhee, I., Barnes, P.J., Giembycz, M.A. and Newton, R. (2001) Adenosine 3',5'-cyclic monophosphate (cAMP)-dependent inhibition of IL-5 from human T lymphocytes is not mediated by the cAMP-dependent protein kinase A. The Journal of Immunology, 167 (4), 2074-2080. (doi:10.4049/?jimmunol.167.4.2074). (PMID:11489990)

Record type: Article

Abstract

IL-5 is implicated in the pathogenesis of asthma and is predominantly released from T lymphocytes of the Th2 phenotype. In anti-CD3 plus anti-CD28-stimulated PBMC, albuterol, isoproterenol, rolipram, PGE2, forskolin, cholera toxin, and the cAMP analog, 8-bromoadenosine cAMP (8-Br-cAMP) all inhibited the release of IL-5 and lymphocyte proliferation. Although all of the above compounds share the ability to increase intracellular cAMP levels and activate protein kinase (PK) A, the PKA inhibitor H-89 failed to ablate the inhibition of IL-5 production mediated by 8-Br-cAMP, rolipram, forskolin, or PGE2. Similarly, H-89 had no effect on the cAMP-mediated inhibition of lymphocyte proliferation. Significantly, these observations occurred at a concentration of H-89 (3 ?M) that inhibited both PKA activity and CREB phosphorylation in intact cells. Additional studies showed that the PKA inhibitors H-8, 8-(4-chlorophenylthio) adenosine-3?,5?-cyclic monophosphorothioate Rp isomer, and a myristolated PKA inhibitor peptide also failed to block the 8-Br-cAMP-mediated inhibition of IL-5 release from PBMC. Likewise, a role for PKG was considered unlikely because both activators and inhibitors of this enzyme had no effect on IL-5 release. Western blotting identified Rap1, a downstream target of the cAMP-binding proteins, exchange protein directly activated by cAMP/cAMP-guanine nucleotide exchange factors 1 and 2, in PBMC. However, Rap1 activation assays revealed that this pathway is also unlikely to be involved in the cAMP-mediated inhibition of IL-5. Taken together, these results indicate that cAMP-elevating agents inhibit IL-5 release from PBMC by a novel cAMP-dependent mechanism that does not involve the activation of PKA

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Published date: 15 August 2001
Organisations: Faculty of Medicine

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Local EPrints ID: 374423
URI: http://eprints.soton.ac.uk/id/eprint/374423
ISSN: 0022-1767
PURE UUID: e2cd3452-47a5-4f2a-8360-efe62687b5d0
ORCID for K.J. Staples: ORCID iD orcid.org/0000-0003-3844-6457

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Date deposited: 16 Feb 2015 14:42
Last modified: 03 Dec 2019 01:46

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Author: K.J. Staples ORCID iD
Author: M. Bergmann
Author: K. Tomita
Author: M.D. Houslay
Author: I. McPhee
Author: P.J. Barnes
Author: M.A. Giembycz
Author: R. Newton

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