title: Induction of activator protein (AP)-1 and nuclear factor-kappaB by CD28 stimulation involves both phosphatidylinositol 3-kinase and acidic sphingomyelinase signals.
abstract: A major obstacle in understanding the signaling events that follow CD28 receptor ligation arises from the fact that CD28 acts as a costimulus to TCR engagement, making it difficult to assess the relative contribution of CD28 signals as distinct from those of the TCR. To overcome this problem, we have exploited the observation that activated human T cell blasts can be stimulated via the CD28 surface molecule in the absence of antigenic challenge; thus, we have been able to observe the response of normal T cells to CD28 activation in isolation. Using this system, we observed that CD28 stimulation by B7-transfected CHO cells induced a proliferative response in T cells that was not accompanied by measurable IL-2 production. However, subsequent analysis of transcription factor generation revealed that B7 stimulation induced both activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) complexes, but not NF-AT. In contrast, engagement of the TCR by class II MHC/superantigen, either with or without CD28 ligation, resulted in the induction of NF-AT, AP-1, and NF-kappaB as well as IL-2 production. Using selective inhibitors, we investigated the signaling pathways involved in the CD28-mediated induction of AP-1 and NF-kappaB. This revealed that NF-kappaB generation was sensitive to chloroquine, an inhibitor of acidic sphingomyelinase, but not to the phosphatidylinositol 3-kinase inhibitor, wortmannin. In contrast, AP-1 generation was inhibited by wortmannin and was also variably sensitive to chloroquine. These data suggest that in activated normal T cells, CD28-derived signals can stimulate proliferation at least in part via NF-kappaB and AP-1 generation, and that this response uses both acidic sphingomyelinase and phosphatidylinositol 3-kinase-linked pathways.
authors.affiliation: Bath Institute for Rheumatic Diseases and School of Pharmacy and Pharmacology, University of Bath, United Kingdom.
authors.forename: C E
authors.initials: CE
authors.lastname: Edmead
authors.forename: Y I
authors.initials: YI
authors.lastname: Patel
authors.forename: A
authors.initials: A
authors.lastname: Wilson
authors.forename: G
authors.initials: G
authors.lastname: Boulougouris
authors.forename: N D
authors.initials: ND
authors.lastname: Hall
authors.forename: S G
authors.initials: SG
authors.lastname: Ward
authors.forename: D M
authors.initials: DM
authors.lastname: Sansom
mesh_terms: D000818:Animals; D018122:B7-1 Antigen / Q000235:genetics / Q000378:metabolism; D018106:CD28 Antigens / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D004789:Enzyme Activation; D006801:Humans; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics; D008213:Lymphocyte Activation; D016328:NF-kappa B / Q000096:biosynthesis*; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D013108:Sphingomyelin Phosphodiesterase / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000096:biosynthesis*; D014157:Transcription Factors / Q000096:biosynthesis; D014162:Transfection
chemical_list: D018122:B7-1 Antigen; D018106:CD28 Antigens; D004268:DNA-Binding Proteins; D007376:Interleukin-2; D016328:NF-kappa B; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D013108:Sphingomyelin Phosphodiesterase