Biphasic control of nuclear factor‐χB activation by the T cell receptor complex: role of tumor necrosis factor α

The regulation of nuclear factor (NF)‐χB activation by the T cell receptor (TcR)/CD3 complex in primary human T cells has been studied at various times after activation. Only p50 NF‐χB protein bound the χB element of interleukin‐2 receptor (IL‐2R) α chain promoter on resting T cells. However, immediately after TcR/CD3 cross‐linking (after approximately 1 h; immediate) binding of p50.p65 heterodimers was observed. p50.c‐rel heterodimers were also detected bound to this sequence at early time points (7–16 h; early), and both remained active at later time points (40 h; late) after activation. This regulation takes place mainly at the level of nuclear translocation of p65 and c‐rel, at immediate and early time points. Activation also induced c‐rel and p105/p50 mRNA synthesis, but not p65 mRNA whose expression was constitutive. Interestingly, all those early and late events, but not the immediate ones, were inhibited by a neutralizing anti‐tumor necrosis factor α (TNF‐α) monoclonal antibody. Similarly, cycloheximide prevented the p65 and c‐rel translocation and consequent formation of active binding heterodimers, at early and late times. Cyclosporin A impaired not only early and late, but also immediate events; however, addition of TNF‐α prevented all inhibition. These results indicate that the regulation of NF‐χB activation during T cell activation by TcR/CD3 signals is biphasic: TcR/CD3 triggers its immediate translocation, which is transient if no TNF‐α is present. TNF‐α, therefore, emerges as the main factor responsible for a second phase of NF‐χB regulation, controlling both translocation of p65 and c‐rel, and new mRNA synthesis for c‐rel and p105/p50.

[1]  P. Vassalli,et al.  Isolation of B and T lymphocytes by nylon fiber columns. , 2008, Tissue antigens.

[2]  M. Fresno,et al.  Control of T lymphocyte activation and IL-2 receptor expression by endogenously secreted lymphokines. , 1994, Journal of immunology.

[3]  I. Verma,et al.  Enhanced I kappa B alpha degradation is responsible for constitutive NF-kappa B activity in mature murine B-cell lines , 1994, Molecular and cellular biology.

[4]  W. Greene,et al.  Autoregulation of the NF-kappa B transactivator RelA (p65) by multiple cytoplasmic inhibitors containing ankyrin motifs. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Dan R. Littman,et al.  Signal transduction by lymphocyte antigen receptors , 1994, Cell.

[6]  L. Glimcher,et al.  Regulation of cytokine gene expression in T helper cell subsets. , 1994, Journal of immunology.

[7]  N. Rice,et al.  In vivo control of NF‐kappa B activation by I kappa B alpha. , 1993, The EMBO journal.

[8]  A. Israël,et al.  Promoter analysis of the gene encoding the I kappa B‐alpha/MAD3 inhibitor of NF‐kappa B: positive regulation by members of the rel/NF‐kappa B family. , 1993, The EMBO journal.

[9]  A. Baldwin,et al.  The I kappa B proteins: multifunctional regulators of Rel/NF-kappa B transcription factors. , 1993, Genes & development.

[10]  T. Wirth,et al.  Distinct NF-κB/Rel transcription factors are responsible for tissue-specific and inducible gene activation , 1993, Nature.

[11]  C. Rosen,et al.  NF-kappa B subunit-specific regulation of the interleukin-8 promoter , 1993, Molecular and cellular biology.

[12]  David Baltimore,et al.  Regulation of the NF-?B/rel transcription factor and I?B inhibitor system , 1993 .

[13]  W C Greene,et al.  NF-kappa B controls expression of inhibitor I kappa B alpha: evidence for an inducible autoregulatory pathway. , 1993, Science.

[14]  G. Franzoso,et al.  Mutual regulation of the transcriptional activator NF-kappa B and its inhibitor, I kappa B-alpha. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[15]  A. Sica,et al.  The interleukin 2 CD28-responsive complex contains at least three members of the NF kappa B family: c-Rel, p50, and p65. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[16]  D. Longo,et al.  Kappa B site-dependent activation of the interleukin-2 receptor alpha-chain gene promoter by human c-Rel , 1992, Molecular and cellular biology.

[17]  P. Askenase,et al.  Molecular regulation of tumor necrosis factor-alpha and lymphotoxin production in T cells. Inhibition by prostaglandin E2. , 1992, Journal of Biological Chemistry.

[18]  A. Israël,et al.  NF-κB and related proteins: Rel/dorsal homologies meet ankyrin-like repeats , 1992 .

[19]  W. Greene,et al.  The 65-kDa subunit of human NF-kappa B functions as a potent transcriptional activator and a target for v-Rel-mediated repression. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[20]  G. Nabel,et al.  Distinct combinations of NF-kappa B subunits determine the specificity of transcriptional activation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[21]  P. Kourilsky,et al.  The characterization of the promoter of the gene encoding the p50 subunit of NF‐kappa B indicates that it participates in its own regulation. , 1992, The EMBO journal.

[22]  L. Aarden,et al.  Activation of interleukin-2 gene transcription via the T-cell surface molecule CD28 is mediated through an NF-kB-like response element. , 1991, The Journal of biological chemistry.

[23]  A. Rao,et al.  Physiologic activation of T cells via the T cell receptor induces NF-kappa B. , 1991, Journal of immunology.

[24]  P. Baeuerle.  The inducible transcription activator NF-κB: regulation by distinct protein subunits , 1991 .

[25]  W. M. Weaver,et al.  Differential regulation of lymphotoxin and tumor necrosis factor genes in human T lymphocytes. , 1991, The Journal of biological chemistry.

[26]  C. Scheidereit,et al.  Cloning of the DNA-binding subunit of human nuclear factor kappa B: the level of its mRNA is strongly regulated by phorbol ester or tumor necrosis factor alpha. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M. Korner,et al.  Nuclear factor kappa B activates proenkephalin transcription in T lymphocytes , 1991, Molecular and cellular biology.

[28]  C. Scheidereit,et al.  Maintenance of NF-kappa B activity is dependent on protein synthesis and the continuous presence of external stimuli , 1991, Molecular and cellular biology.

[29]  W C Greene,et al.  NF-kappa B: a family of inducible and differentially expressed enhancer-binding proteins in human T cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Steinman,et al.  The effect of immunosuppressive agents on the induction of nuclear factors that bind to sites on the interleukin 2 promoter , 1990, The Journal of experimental medicine.

[31]  W. Greene,et al.  The v-rel oncogene encodes a κB enhancer binding protein that inhibits NF-κB function , 1990, Cell.

[32]  D. Thomas,et al.  Stimulation of a human T-cell clone with anti-CD3 or tumor necrosis factor induces NF-kappa B translocation but not human immunodeficiency virus 1 enhancer-dependent transcription. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[33]  G. Nolan,et al.  Cloning of the p50 DNA binding subunit of NF-κB: Homology to rel and dorsal , 1990, Cell.

[34]  A. Israël,et al.  The DNA binding subunit of NF-κB is identical to factor KBF1 and homologous to the rel oncogene product , 1990, Cell.

[35]  L. Hennighausen,et al.  Inducible nuclear factor binding to the kappa B elements of the human immunodeficiency virus enhancer in T cells can be blocked by cyclosporin A in a signal-dependent manner , 1990, Journal of virology.

[36]  S. Gerondakis,et al.  Murine c-rel transcription is rapidly induced in T-cells and fibroblasts by mitogenic agents and the phorbol ester 12-O-tetradecanoylphorbol-13-acetate. , 1990, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[37]  P. Baeuerle,et al.  NF-kappa B as inducible transcriptional activator of the granulocyte-macrophage colony-stimulating factor gene , 1990, Molecular and cellular biology.

[38]  W. Leonard,et al.  The same target sequences are differentially important for activation of the interleukin 2 receptor alpha-chain gene in two distinct T-cell lines. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[39]  S. L. Cross,et al.  Delineation of an enhancerlike positive regulatory element in the interleukin-2 receptor alpha-chain gene , 1990, Molecular and cellular biology.

[40]  G. Nabel,et al.  Regulation of the IL-2 receptor alpha-gene. Interaction of a kappa B binding protein with cell-specific transcription factors. , 1989, Journal of immunology.

[41]  W. Schaffner,et al.  Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. , 1989, Nucleic acids research.

[42]  W. Greene,et al.  Tumor necrosis factor-alpha activation of the IL-2 receptor-alpha gene involves the induction of kappa B-specific DNA binding proteins. , 1989, Journal of immunology.

[43]  M. Siekevitz,et al.  Kappa B-specific DNA binding proteins: role in the regulation of human interleukin-2 gene expression. , 1989, Science.

[44]  G. Nabel,et al.  Tumor necrosis factor alpha and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor kappa B. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[45]  W. Greene,et al.  Tumor necrosis factor alpha induces proteins that bind specifically to kappa B-like enhancer elements and regulate interleukin 2 receptor alpha-chain gene expression in primary human T lymphocytes. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[46]  G. Crabtree.  Contingent genetic regulatory events in T lymphocyte activation. , 1989, Science.

[47]  W. Greene,et al.  Regulation of interleukin 2 receptor alpha subunit (Tac or CD25 antigen) gene expression: binding of inducible nuclear proteins to discrete promoter sequences correlates with transcriptional activation. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[48]  C. Y. Wang,et al.  Production of tumor necrosis factor/cachectin by human T cell lines and peripheral blood T lymphocytes stimulated by phorbol myristate acetate and anti-CD3 antibody , 1988, The Journal of experimental medicine.

[49]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[50]  David Baltimore,et al.  Inducibility of κ immunoglobulin enhancer-binding protein NF-κB by a posttranslational mechanism , 1986, Cell.

[51]  D. Cantrell,et al.  The interleukin-2 T-cell system: a new cell growth model. , 1984, Science.

[52]  E. Reinherz,et al.  Triggering of the T3-Ti antigen-receptor complex results in clonal T-cell proliferation through an interleukin 2-dependent autocrine pathway. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[53]  N. Sigal,et al.  Cyclosporin A, FK-506, and Rapamycin: Pharmacologic Probes of Lymphocyte Signal Transduction , 1992 .

[54]  G. Crabtree,et al.  Transmission of signals from the T lymphocyte antigen receptor to the genes responsible for cell proliferation and immune function: the missing link. , 1990, Annual review of immunology.

[55]  T. Mustelin,et al.  Molecular events mediating T cell activation. , 1990, Advances in immunology.

[56]  K. Smith,et al.  The interleukin 2 receptor. , 1989, Annual review of cell biology.

[57]  T. Taniguchi,et al.  Involvement of a common transcription factor in the regulated expression of IL-2 and IL-2 receptor genes. , 1989, International immunology.

[58]  J. D. de Vries,et al.  Antigen‐specific cytotoxic T cell and antigen‐specific proliferating T cell clones can be induced to cytolytic activity by monoclonal antibodies against T3 , 1985, European journal of immunology.