{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T02:00:28Z","timestamp":1791252028223,"version":"4.1.0"},"reference-count":56,"publisher":"American Society of Hematology","issue":"3","content-domain":{"domain":["ashpublications.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2003,2,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cyclic adenosine monophosphate (cAMP) is a negative regulator of T-cell activation. However, the effects of cAMP on signaling pathways that regulate cytokine production and cell cycle progression remain unclear. Here, using primary human T lymphocytes in which endogenous cAMP was increased by the use of forskolin and 3-isobutyl-1-methylxanthine (IBMX), we show that increase of cAMP resulted in inhibition of T-cell receptor (TCR)\/CD3 plus CD28\u2013mediated T-cell activation and cytokine production and blockade of cell cycle progression at the G1 phase. Increase of cAMP inhibited Ras activation and phosphorylation of mitogen-induced extracellular kinase (MEK) downstream targets extracellular signal\u2013related kinase 1\/2 (ERK1\/2) and phosphatidylinositol-3-kinase (PI3K) downstream target protein kinase B (PKB; c-Akt). These functional and biochemical events were secondary to the impaired activation of ZAP-70 and phosphorylation of LAT and did not occur when cells were stimulated with phorbol ester, which bypasses the TCR proximal signaling events and activates Ras. Increase of cAMP also inhibited activation of Rap1 mediated by TCR\/CD3 plus CD28. Importantly, inhibition of Rap1 activation by cAMP was also observed when cells were stimulated with phorbol ester, although under these conditions Ras was activated and cells progressed into the cell cycle. Thus, TCR plus CD28\u2013mediated activation of ERK1\/2 and PKB, cytokine production, and cell cycle progression, all of which are inhibited by cAMP, require activation of Ras but not Rap1. These results indicate that signals that regulate cAMP levels after encounter of T cells by antigen will likely determine the functional fate toward clonal expansion or repression of primary T-cell responses.<\/jats:p>","DOI":"10.1182\/blood-2002-06-1665","type":"journal-article","created":{"date-parts":[[2003,1,15]],"date-time":"2003-01-15T21:05:10Z","timestamp":1042664710000},"page":"998-1006","update-policy":"https:\/\/doi.org\/10.1182\/blood.2019cm0000","source":"Crossref","is-referenced-by-count":42,"title":["cAMP inhibits both Ras and Rap1 activation in primary human T lymphocytes, but only Ras inhibition correlates with blockade of cell cycle progression"],"prefix":"10.1182","volume":"101","author":[{"given":"Thomas","family":"Grader-Beck","sequence":"first","affiliation":[{"name":"From the Department of Adult Oncology, Dana-Farber Cancer Institute; Division of Medical Oncology, Brigham and Women's Hospital; and Department of Medicine, Harvard Medical School, Boston, MA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andre A. F. L.","family":"van Puijenbroek","sequence":"additional","affiliation":[{"name":"From the Department of Adult Oncology, Dana-Farber Cancer Institute; Division of Medical Oncology, Brigham and Women's Hospital; and Department of Medicine, Harvard Medical School, Boston, MA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lee M.","family":"Nadler","sequence":"additional","affiliation":[{"name":"From the Department of Adult Oncology, Dana-Farber Cancer Institute; Division of Medical Oncology, Brigham and Women's Hospital; and Department of Medicine, Harvard Medical School, Boston, MA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vassiliki A.","family":"Boussiotis","sequence":"additional","affiliation":[{"name":"From the Department of Adult Oncology, Dana-Farber Cancer Institute; Division of Medical Oncology, Brigham and Women's Hospital; and Department of Medicine, Harvard Medical School, Boston, MA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"234","reference":[{"key":"2019111610411410500_B1","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/S0162-3109(97)00071-4","article-title":"A selective type 4 phosphodiesterase inhibitor, T-440, modulates intracellular cyclic AMP level and interleukin-2 production of Jurkat cells.","volume":"38","author":"Kaminuma","year":"1998","journal-title":"Immunopharmacology."},{"key":"2019111610411410500_B2","doi-asserted-by":"crossref","first-page":"8991","DOI":"10.1073\/pnas.95.15.8991","article-title":"Cloning and characterization of a cAMP-specific cyclic nucleotide phosphodiesterase.","volume":"95","author":"Soderling","year":"1998","journal-title":"Proc Natl Acad Sci U S A."},{"key":"2019111610411410500_B3","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/0968-0004(89)90046-7","article-title":"The cyclic AMP-mediated stimulation of cell proliferation.","volume":"14","author":"Dumont","year":"1989","journal-title":"Trends Biochem Sci."},{"key":"2019111610411410500_B4","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1002\/jcp.1041370327","article-title":"cAMP-mediated growth inhibition of a B-lymphoid precursor cell line Reh is associated with an early transient delay in G2\/M, followed by an accumulation of cells in G1.","volume":"137","author":"Blomhoff","year":"1988","journal-title":"J Cell Physiol."},{"key":"2019111610411410500_B5","doi-asserted-by":"crossref","first-page":"25842","DOI":"10.1074\/jbc.274.36.25842","article-title":"Differential effects of cAMP in neurons and astrocytes. Role of B-raf.","volume":"274","author":"Dugan","year":"1999","journal-title":"J Biol Chem."},{"key":"2019111610411410500_B6","doi-asserted-by":"crossref","first-page":"2845","DOI":"10.4049\/jimmunol.148.9.2845","article-title":"Prostaglandin E2 and other cyclic AMP-elevated agents modulate IL-2 and IL-2R alpha gene expression at multiple levels.","volume":"148","author":"Anastasiou","year":"1992","journal-title":"J Immunol."},{"key":"2019111610411410500_B7","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1084\/jem.179.3.931","article-title":"Interleukin 2 transcription factors as molecular targets of cAMP inhibition: delayed inhibition kinetics and combinatorial transcription roles.","volume":"179","author":"Chen","year":"1994","journal-title":"J Exp Med."},{"key":"2019111610411410500_B8","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.4049\/jimmunol.164.2.1117","article-title":"Therapeutic potential of phosphodiesterase-4 and -3 inhibitors in Th1-mediated autoimmune diseases.","volume":"164","author":"Bielekova","year":"2000","journal-title":"J. Immunol."},{"key":"2019111610411410500_B9","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1016\/0167-5699(88)91220-0","article-title":"The adenylate cyclase-cAMP-protein kinase A pathway and regulation of the immune response.","volume":"9","author":"Kammer","year":"1988","journal-title":"Immunol Today."},{"key":"2019111610411410500_B10","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1126\/science.8272870","article-title":"Location of cAMP-dependent protein kinase type I with the TCR-CD3 complex.","volume":"263","author":"Skalhegg","year":"1994","journal-title":"Science."},{"key":"2019111610411410500_B11","doi-asserted-by":"crossref","first-page":"6676","DOI":"10.1073\/pnas.90.14.6676","article-title":"Human immunodeficiency virus proteins induce the inhibitory cAMP\/protein kinase A pathway in normal lymphocytes.","volume":"90","author":"Hofmann","year":"1993","journal-title":"Proc Natl Acad Sci U S A."},{"key":"2019111610411410500_B12","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1096\/fasebj.12.10.855","article-title":"Protein kinase A type I antagonist restores immune responses of T cells from HIV-infected patients.","volume":"12","author":"Aandahl","year":"1998","journal-title":"FASEB J."},{"key":"2019111610411410500_B13","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.4049\/jimmunol.162.2.1178","article-title":"Increased activation of protein kinase A type I contributes to the T cell deficiency in common variable immunodeficiency.","volume":"162","author":"Aukrust","year":"1999","journal-title":"J Immunol."},{"key":"2019111610411410500_B14","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1038\/73144","article-title":"p27kip1 functions as an anergy factor inhibiting IL-2 transcription and clonal expansion of alloreactive human and murine helper T lymphocytes.","volume":"6","author":"Boussiotis","year":"2000","journal-title":"Nat Med."},{"key":"2019111610411410500_B15","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/379081a0","article-title":"Defective thymocyte proliferation and IL-2 production in transgenic mice expressing a dominant-negative form of CREB.","volume":"379","author":"Barton","year":"1996","journal-title":"Nature."},{"key":"2019111610411410500_B16","doi-asserted-by":"crossref","first-page":"6631","DOI":"10.4049\/jimmunol.163.12.6631","article-title":"The -180 site of the IL-2 promoter is the target of CREB\/CREM binding in T cell anergy.","volume":"163","author":"Powell","year":"1999","journal-title":"J Immunol."},{"key":"2019111610411410500_B17","doi-asserted-by":"crossref","first-page":"848","DOI":"10.1126\/science.283.5403.848","article-title":"CD3- and CD28-dependent induction of PDE7 required for T cell activation.","volume":"283","author":"Li","year":"1999","journal-title":"Science."},{"key":"2019111610411410500_B18","doi-asserted-by":"crossref","first-page":"2729","DOI":"10.4049\/jimmunol.168.6.2729","article-title":"CD28 costimulation mediates down-regulation of p27(kip1) and cell cycle progression by activation of the PI3K\/PKB signaling pathway in primary human T cells.","volume":"168","author":"Appleman","year":"2002","journal-title":"J Immunol."},{"key":"2019111610411410500_B19","doi-asserted-by":"crossref","first-page":"4944","DOI":"10.1074\/jbc.273.9.4944","article-title":"Costimulation through CD28 suppresses T cell receptor-dependent activation of the Ras-like small GTPase Rap1 in human T lymphocytes.","volume":"273","author":"Reedquist","year":"1998","journal-title":"J Bio. 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Rooij","year":"1998","journal-title":"Nature."},{"key":"2019111610411410500_B34","doi-asserted-by":"crossref","first-page":"13278","DOI":"10.1073\/pnas.95.22.13278","article-title":"A Rap guanine nucleotide exchange factor enriched highly in the basal ganglia.","volume":"95","author":"Kawasaki","year":"1998","journal-title":"Proc Natl Acad Sci U S A."},{"key":"2019111610411410500_B35","doi-asserted-by":"crossref","first-page":"30301","DOI":"10.1074\/jbc.M103530200","article-title":"Role of the CDC25 homology domain of phospholipase Cepsilon in amplification of Rap1-dependent signaling.","volume":"276","author":"Jin","year":"2001","journal-title":"J Biol Chem."},{"key":"2019111610411410500_B36","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1038\/346719a0","article-title":"Stimulation of p21ras upon T-cell activation.","volume":"346","author":"Downward","year":"1990","journal-title":"Nature."},{"key":"2019111610411410500_B37","first-page":"933","article-title":"Signal transduction via Ras.","volume":"379","author":"Wittinghofer","year":"1998","journal-title":"Biol Chem."},{"key":"2019111610411410500_B38","doi-asserted-by":"crossref","first-page":"4435","DOI":"10.1128\/MCB.14.7.4435","article-title":"GRB2 and phospholipase C-gamma 1 associate with a 36- to 38-kilodalton phosphotyrosine protein after T-cell receptor stimulation.","volume":"14","author":"Sieh","year":"1994","journal-title":"Mol Cell Biol."},{"key":"2019111610411410500_B39","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/S0092-8674(00)80901-0","article-title":"LAT: the ZAP-70 tyrosine kinase substrate that links T cell receptor to cellular activation.","volume":"92","author":"Zhang","year":"1998","journal-title":"Cell."},{"key":"2019111610411410500_B40","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1038\/369411a0","article-title":"Requirement for Ras in Raf activation is overcome by targeting Raf to the plasma membrane.","volume":"369","author":"Leevers","year":"1994","journal-title":"Nature."},{"key":"2019111610411410500_B41","doi-asserted-by":"crossref","first-page":"1463","DOI":"10.1126\/science.7811320","article-title":"Activation of Ras as a result of recruitment to the plasma membrane.","volume":"264","author":"Stokoe","year":"1994","journal-title":"Science."},{"key":"2019111610411410500_B42","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1038\/357602a0","article-title":"Involvement of p21ras in activation of extracellular signal-regulated kinase 2.","volume":"357","author":"de Vries-Smits","year":"1992","journal-title":"Nature."},{"key":"2019111610411410500_B43","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1084\/jem.193.4.497","article-title":"Activation of the COOH-terminal src kinase (Csk) by cAMP-dependent protein kinase inhibits signaling through the T cell receptor.","volume":"193","author":"Vang","year":"2001","journal-title":"J Exp Med."},{"key":"2019111610411410500_B44","doi-asserted-by":"crossref","first-page":"5882","DOI":"10.1128\/MCB.19.9.5882","article-title":"Protein kinase A-dependent and -independent signaling pathways contribute to cyclic AMP-stimulated proliferation.","volume":"19","author":"Cass","year":"1999","journal-title":"Mol Cell Biol."},{"key":"2019111610411410500_B45","doi-asserted-by":"crossref","first-page":"6746","DOI":"10.1128\/MCB.15.12.6746","article-title":"Identification of Rap1 as a Target for the crk SH3 domain binding guanine nucleotide-releasing factor C3G.","volume":"15","author":"Gotoh","year":"1995","journal-title":"Mol Cell Biol."},{"key":"2019111610411410500_B46","doi-asserted-by":"crossref","first-page":"2275","DOI":"10.1126\/science.282.5397.2275","article-title":"A family of cAMP-binding proteins that directly activate Rap1.","volume":"282","author":"Kawasaki","year":"1998","journal-title":"Science."},{"key":"2019111610411410500_B47","doi-asserted-by":"crossref","first-page":"554","DOI":"10.1038\/375554a0","article-title":"The 2.2 A crystal structure of the Ras-binding domain of the serine\/threonine kinase c-Raf1 in complex with Rap1A and a GTP analogue.","volume":"375","author":"Nassar","year":"1995","journal-title":"Nature."},{"key":"2019111610411410500_B48","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1126\/science.278.5335.124","article-title":"Maintenance of human T cell anergy: blocking of IL-2 gene transcription by activated Rap1.","volume":"278","author":"Boussiotis","year":"1997","journal-title":"Science."},{"key":"2019111610411410500_B49","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1126\/science.7694367","article-title":"Inhibition by cAMP of Ras-dependent activation of Raf.","volume":"262","author":"Cook","year":"1993","journal-title":"Science."},{"key":"2019111610411410500_B50","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/S0092-8674(00)80184-1","article-title":"cAMP activates MAP kinase and Elk-1 through a B-Raf-and Rap1-dependent pathway.","volume":"89","author":"Vossler","year":"1997","journal-title":"Cell."},{"key":"2019111610411410500_B51","doi-asserted-by":"crossref","first-page":"3671","DOI":"10.1128\/MCB.21.11.3671-3683.2001","article-title":"Cyclic AMP-mediated inhibition of cell growth requires the small protein Rap1.","volume":"21","author":"Schmitt","year":"2001","journal-title":"Mol Cell Biol."},{"key":"2019111610411410500_B52","doi-asserted-by":"crossref","first-page":"1991","DOI":"10.1210\/endo.139.4.5880","article-title":"Differential effects of cyclic adenosine 3\u2032,5\u2032-monophosphate on p70 ribosomal S6 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