Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. Jul 1997; 17(7): 4051–4058.
PMCID: PMC232258

CD28 mediates transcriptional upregulation of the interleukin-2 (IL-2) promoter through a composite element containing the CD28RE and NF-IL-2B AP-1 sites.


Mutagenesis studies have demonstrated the requirement for the CD28-responsive element (CD28RE) within the interleukin-2 (IL-2) promoter for transcriptional upregulation by CD28. Here, we demonstrate that CD28 responsiveness is conferred by a composite element containing both the CD28RE and the NF-IL-2B AP-1 sites (RE/AP). Mutations at either site within the RE/AP composite element abolish activity. The RE/AP composite element is a site for signal integration within the IL-2 promoter, since its activation is dependent on at least two separate signalling pathways being activated, through the T-cell receptor, CD28, and/or phorbol myristate acetate. Activation is maximal when all three signals occur simultaneously. By using a panel of CD28 cytoplasmic domain mutants, it was found that the transcriptional activation of the RE/AP composite element correlates exactly with the pattern of IL-2 secretion induced by these mutants upon stimulation. Similar to the upregulation of IL-2 secretion, the transcriptional upregulation of the RE/AP composite element by CD28 is FK506 insensitive. The pattern of activation of the RE/AP composite element is different from that observed for either an NFAT or consensus AP-1 site, implying that RE/AP represents a unique element. Using gel shift analysis, we demonstrate that stimulation by CD28 induces the association of the NF-kappaB family member c-Rel to the CD28RE within the RE/AP composite element. The transcriptional upregulation of IL-2 by CD28 appears, therefore, to be mediated through the RE/AP composite element, involving the association of c-Rel with the CD28RE.

Full Text

The Full Text of this article is available as a PDF (1.0M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Civil A, Bakker A, Rensink I, Doerre S, Aarden LA, Verweij CL. Nuclear appearance of a factor that binds the CD28 response element within the interleukin-2 enhancer correlates with interleukin-2 production. J Biol Chem. 1996 Apr 5;271(14):8321–8327. [PubMed]
  • Civil A, Geerts M, Aarden LA, Verweij CL. Evidence for a role of CD28RE as a response element for distinct mitogenic T cell activation signals. Eur J Immunol. 1992 Nov;22(11):3041–3043. [PubMed]
  • Fraser JD, Irving BA, Crabtree GR, Weiss A. Regulation of interleukin-2 gene enhancer activity by the T cell accessory molecule CD28. Science. 1991 Jan 18;251(4991):313–316. [PubMed]
  • Fraser JD, Weiss A. Regulation of T-cell lymphokine gene transcription by the accessory molecule CD28. Mol Cell Biol. 1992 Oct;12(10):4357–4363. [PMC free article] [PubMed]
  • Ghosh P, Tan TH, Rice NR, Sica A, Young HA. The interleukin 2 CD28-responsive complex contains at least three members of the NF kappa B family: c-Rel, p50, and p65. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1696–1700. [PMC free article] [PubMed]
  • Good L, Maggirwar SB, Sun SC. Activation of the IL-2 gene promoter by HTLV-I tax involves induction of NF-AT complexes bound to the CD28-responsive element. EMBO J. 1996 Jul 15;15(14):3744–3750. [PMC free article] [PubMed]
  • Harding FA, McArthur JG, Gross JA, Raulet DH, Allison JP. CD28-mediated signalling co-stimulates murine T cells and prevents induction of anergy in T-cell clones. Nature. 1992 Apr 16;356(6370):607–609. [PubMed]
  • Himes SR, Coles LS, Reeves R, Shannon MF. High mobility group protein I(Y) is required for function and for c-Rel binding to CD28 response elements within the GM-CSF and IL-2 promoters. Immunity. 1996 Nov;5(5):479–489. [PubMed]
  • Hodge MR, Ranger AM, Charles de la Brousse F, Hoey T, Grusby MJ, Glimcher LH. Hyperproliferation and dysregulation of IL-4 expression in NF-ATp-deficient mice. Immunity. 1996 Apr;4(4):397–405. [PubMed]
  • Hughes CC, Pober JS. Transcriptional regulation of the interleukin-2 gene in normal human peripheral blood T cells. Convergence of costimulatory signals and differences from transformed T cells. J Biol Chem. 1996 Mar 8;271(10):5369–5377. [PubMed]
  • Jain J, Loh C, Rao A. Transcriptional regulation of the IL-2 gene. Curr Opin Immunol. 1995 Jun;7(3):333–342. [PubMed]
  • Jain J, Valge-Archer VE, Rao A. Analysis of the AP-1 sites in the IL-2 promoter. J Immunol. 1992 Feb 15;148(4):1240–1250. [PubMed]
  • Jenkins MK. The ups and downs of T cell costimulation. Immunity. 1994 Sep;1(6):443–446. [PubMed]
  • June CH, Bluestone JA, Nadler LM, Thompson CB. The B7 and CD28 receptor families. Immunol Today. 1994 Jul;15(7):321–331. [PubMed]
  • Kang SM, Beverly B, Tran AC, Brorson K, Schwartz RH, Lenardo MJ. Transactivation by AP-1 is a molecular target of T cell clonal anergy. Science. 1992 Aug 21;257(5073):1134–1138. [PubMed]
  • Köntgen F, Grumont RJ, Strasser A, Metcalf D, Li R, Tarlinton D, Gerondakis S. Mice lacking the c-rel proto-oncogene exhibit defects in lymphocyte proliferation, humoral immunity, and interleukin-2 expression. Genes Dev. 1995 Aug 15;9(16):1965–1977. [PubMed]
  • Lai JH, Horvath G, Subleski J, Bruder J, Ghosh P, Tan TH. RelA is a potent transcriptional activator of the CD28 response element within the interleukin 2 promoter. Mol Cell Biol. 1995 Aug;15(8):4260–4271. [PMC free article] [PubMed]
  • LaSalle JM, Hafler DA. T cell anergy. FASEB J. 1994 Jun;8(9):601–608. [PubMed]
  • Meyer CF, Wang X, Chang C, Templeton D, Tan TH. Interaction between c-Rel and the mitogen-activated protein kinase kinase kinase 1 signaling cascade in mediating kappaB enhancer activation. J Biol Chem. 1996 Apr 12;271(15):8971–8976. [PubMed]
  • Pierce JW, Jamieson CA, Ross JL, Sen R. Activation of IL-2 receptor alpha-chain gene by individual members of the rel oncogene family in association with serum response factor. J Immunol. 1995 Aug 15;155(4):1972–1980. [PubMed]
  • Rao A. NF-ATp: a transcription factor required for the co-ordinate induction of several cytokine genes. Immunol Today. 1994 Jun;15(6):274–281. [PubMed]
  • Riegel JS, Corthesy B, Flanagan WM, Crabtree GR. Regulation of the interleukin-2 gene. Chem Immunol. 1992;51:266–298. [PubMed]
  • Rincón M, Flavell RA. AP-1 transcriptional activity requires both T-cell receptor-mediated and co-stimulatory signals in primary T lymphocytes. EMBO J. 1994 Sep 15;13(18):4370–4381. [PMC free article] [PubMed]
  • Rooney JW, Sun YL, Glimcher LH, Hoey T. Novel NFAT sites that mediate activation of the interleukin-2 promoter in response to T-cell receptor stimulation. Mol Cell Biol. 1995 Nov;15(11):6299–6310. [PMC free article] [PubMed]
  • Schreiber E, Matthias P, Müller MM, Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. [PMC free article] [PubMed]
  • Serfling E, Avots A, Neumann M. The architecture of the interleukin-2 promoter: a reflection of T lymphocyte activation. Biochim Biophys Acta. 1995 Sep 19;1263(3):181–200. [PubMed]
  • Sha WC, Liou HC, Tuomanen EI, Baltimore D. Targeted disruption of the p50 subunit of NF-kappa B leads to multifocal defects in immune responses. Cell. 1995 Jan 27;80(2):321–330. [PubMed]
  • Shapiro VS, Mollenauer MN, Greene WC, Weiss A. c-rel regulation of IL-2 gene expression may be mediated through activation of AP-1. J Exp Med. 1996 Nov 1;184(5):1663–1669. [PMC free article] [PubMed]
  • Springer TA. Adhesion receptors of the immune system. Nature. 1990 Aug 2;346(6283):425–434. [PubMed]
  • Su B, Jacinto E, Hibi M, Kallunki T, Karin M, Ben-Neriah Y. JNK is involved in signal integration during costimulation of T lymphocytes. Cell. 1994 Jun 3;77(5):727–736. [PubMed]
  • Truitt KE, Nagel T, Suen LF, Imboden JB. Structural requirements for CD28-mediated costimulation of IL-2 production in Jurkat T cells. J Immunol. 1996 Jun 15;156(12):4539–4541. [PubMed]
  • Umlauf SW, Beverly B, Kang SM, Brorson K, Tran AC, Schwartz RH. Molecular regulation of the IL-2 gene: rheostatic control of the immune system. Immunol Rev. 1993 Jun;133:177–197. [PubMed]
  • Weiss A, Littman DR. Signal transduction by lymphocyte antigen receptors. Cell. 1994 Jan 28;76(2):263–274. [PubMed]
  • Xanthoudakis S, Viola JP, Shaw KT, Luo C, Wallace JD, Bozza PT, Luk DC, Curran T, Rao A. An enhanced immune response in mice lacking the transcription factor NFAT1. Science. 1996 May 10;272(5263):892–895. [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...