Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 Nov 1;102(9):1645–1652. doi: 10.1172/JCI3877

Regulation of an essential innate immune response by the p50 subunit of NF-kappaB.

J Bohuslav 1, V V Kravchenko 1, G C Parry 1, J H Erlich 1, S Gerondakis 1, N Mackman 1, R J Ulevitch 1
PMCID: PMC509112  PMID: 9802878

Abstract

Recognition of bacterial endotoxin (LPS) elicits multiple host responses, including activation of cells of the innate immune system. LPS exposure occurs repeatedly during septicemia, making strict regulation of gene expression necessary. Such regulation might prevent, for example, the continuous production of proinflammatory cytokines such as tumor necrosis factor (TNF), which could lead to severe vascular collapse. Tolerance to LPS is characterized by a diminished production of TNF during prolonged exposure to LPS, and is therefore likely to represent an essential control mechanism during sepsis. In the present study, which uses mice with genetic deletions of the proteins of NF-kappaB complex, we provide data demonstrating that increased expression of the p50 subunit of NF-kappaB directly results in the downregulation of LPS-induced TNF production. This contention is supported by the following observations: (1) tolerance to LPS is not induced in macrophages from p50-/- mice; (2) long-term pretreatment with LPS does not block synthesis of TNF mRNA in p50-/- macrophages (in contrast to wild-type macrophages); (3) ectopic overexpression of p50 reduces transcriptional activation of the murine TNF promoter; and (4) analysis of the four kappaB sites from the murine TNF promoter demonstrates that binding of p50 homodimers to the positively acting kappaB3 element is associated with development of the LPS-tolerant phenotype. Thus, p50 expression plays a key role in the development of LPS tolerance.

Full Text

The Full Text of this article is available as a PDF (853.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boyte W. R., Meals E. A., English B. K. Acquired hyporesponsiveness to bacterial lipopolysaccharide and interferon-gamma in RAW 264.7 macrophages. Shock. 1996 Sep;6(3):218–222. [PubMed] [Google Scholar]
  2. Chiao P. J., Miyamoto S., Verma I. M. Autoregulation of I kappa B alpha activity. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):28–32. doi: 10.1073/pnas.91.1.28. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collart M. A., Baeuerle P., Vassalli P. Regulation of tumor necrosis factor alpha transcription in macrophages: involvement of four kappa B-like motifs and of constitutive and inducible forms of NF-kappa B. Mol Cell Biol. 1990 Apr;10(4):1498–1506. doi: 10.1128/mcb.10.4.1498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cordle S. R., Donald R., Read M. A., Hawiger J. Lipopolysaccharide induces phosphorylation of MAD3 and activation of c-Rel and related NF-kappa B proteins in human monocytic THP-1 cells. J Biol Chem. 1993 Jun 5;268(16):11803–11810. [PubMed] [Google Scholar]
  5. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Drouet C., Shakhov A. N., Jongeneel C. V. Enhancers and transcription factors controlling the inducibility of the tumor necrosis factor-alpha promoter in primary macrophages. J Immunol. 1991 Sep 1;147(5):1694–1700. [PubMed] [Google Scholar]
  7. Frankenberger M., Ziegler-Heitbrock H. W. LPS tolerance in monocytes/macrophages: three 3' cytosins are required in the DNA binding motif for detection of upregulated NF-kappa B p50 homodimers. Immunobiology. 1997 Dec;198(1-3):81–90. doi: 10.1016/s0171-2985(97)80029-0. [DOI] [PubMed] [Google Scholar]
  8. Franzoso G., Bours V., Park S., Tomita-Yamaguchi M., Kelly K., Siebenlist U. The candidate oncoprotein Bcl-3 is an antagonist of p50/NF-kappa B-mediated inhibition. Nature. 1992 Sep 24;359(6393):339–342. doi: 10.1038/359339a0. [DOI] [PubMed] [Google Scholar]
  9. Gerondakis S., Strasser A., Metcalf D., Grigoriadis G., Scheerlinck J. Y., Grumont R. J. Rel-deficient T cells exhibit defects in production of interleukin 3 and granulocyte-macrophage colony-stimulating factor. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3405–3409. doi: 10.1073/pnas.93.8.3405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Glauser M. P., Zanetti G., Baumgartner J. D., Cohen J. Septic shock: pathogenesis. Lancet. 1991 Sep 21;338(8769):732–736. doi: 10.1016/0140-6736(91)91452-z. [DOI] [PubMed] [Google Scholar]
  11. Grigoriadis G., Zhan Y., Grumont R. J., Metcalf D., Handman E., Cheers C., Gerondakis S. The Rel subunit of NF-kappaB-like transcription factors is a positive and negative regulator of macrophage gene expression: distinct roles for Rel in different macrophage populations. EMBO J. 1996 Dec 16;15(24):7099–7107. [PMC free article] [PubMed] [Google Scholar]
  12. Han J., Huez G., Beutler B. Interactive effects of the tumor necrosis factor promoter and 3'-untranslated regions. J Immunol. 1991 Mar 15;146(6):1843–1848. [PubMed] [Google Scholar]
  13. Kravchenko V. V., Pan Z., Han J., Herbert J. M., Ulevitch R. J., Ye R. D. Platelet-activating factor induces NF-kappa B activation through a G protein-coupled pathway. J Biol Chem. 1995 Jun 23;270(25):14928–14934. doi: 10.1074/jbc.270.25.14928. [DOI] [PubMed] [Google Scholar]
  14. Kravchenko V. V., Steinemann S., Kline L., Feng L., Ulevitch R. J. Endotoxin tolerance is induced in Chinese hamster ovary cell lines expressing human CD14. Shock. 1996 Mar;5(3):194–201. doi: 10.1097/00024382-199603000-00005. [DOI] [PubMed] [Google Scholar]
  15. Kunsch C., Ruben S. M., Rosen C. A. Selection of optimal kappa B/Rel DNA-binding motifs: interaction of both subunits of NF-kappa B with DNA is required for transcriptional activation. Mol Cell Biol. 1992 Oct;12(10):4412–4421. doi: 10.1128/mcb.12.10.4412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Köntgen F., Grumont R. J., 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. doi: 10.1101/gad.9.16.1965. [DOI] [PubMed] [Google Scholar]
  17. LaRue K. E., McCall C. E. A labile transcriptional repressor modulates endotoxin tolerance. J Exp Med. 1994 Dec 1;180(6):2269–2275. doi: 10.1084/jem.180.6.2269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mathison J. C., Virca G. D., Wolfson E., Tobias P. S., Glaser K., Ulevitch R. J. Adaptation to bacterial lipopolysaccharide controls lipopolysaccharide-induced tumor necrosis factor production in rabbit macrophages. J Clin Invest. 1990 Apr;85(4):1108–1118. doi: 10.1172/JCI114542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mathison J. C., Wolfson E., Ulevitch R. J. Participation of tumor necrosis factor in the mediation of gram negative bacterial lipopolysaccharide-induced injury in rabbits. J Clin Invest. 1988 Jun;81(6):1925–1937. doi: 10.1172/JCI113540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Parry G. C., Mackman N. A set of inducible genes expressed by activated human monocytic and endothelial cells contain kappa B-like sites that specifically bind c-Rel-p65 heterodimers. J Biol Chem. 1994 Aug 19;269(33):20823–20825. [PubMed] [Google Scholar]
  21. Schmitz M. L., Baeuerle P. A. The p65 subunit is responsible for the strong transcription activating potential of NF-kappa B. EMBO J. 1991 Dec;10(12):3805–3817. doi: 10.1002/j.1460-2075.1991.tb04950.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sha W. C., Liou H. C., Tuomanen E. I., 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. doi: 10.1016/0092-8674(95)90415-8. [DOI] [PubMed] [Google Scholar]
  23. Shakhov A. N., Collart M. A., Vassalli P., Nedospasov S. A., Jongeneel C. V. Kappa B-type enhancers are involved in lipopolysaccharide-mediated transcriptional activation of the tumor necrosis factor alpha gene in primary macrophages. J Exp Med. 1990 Jan 1;171(1):35–47. doi: 10.1084/jem.171.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ulevitch R. J., Tobias P. S. Recognition of endotoxin by cells leading to transmembrane signaling. Curr Opin Immunol. 1994 Feb;6(1):125–130. doi: 10.1016/0952-7915(94)90043-4. [DOI] [PubMed] [Google Scholar]
  25. Urban M. B., Baeuerle P. A. The 65-kD subunit of NF-kappa B is a receptor for I kappa B and a modulator of DNA-binding specificity. Genes Dev. 1990 Nov;4(11):1975–1984. doi: 10.1101/gad.4.11.1975. [DOI] [PubMed] [Google Scholar]
  26. Urban M. B., Schreck R., Baeuerle P. A. NF-kappa B contacts DNA by a heterodimer of the p50 and p65 subunit. EMBO J. 1991 Jul;10(7):1817–1825. doi: 10.1002/j.1460-2075.1991.tb07707.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Virca G. D., Kim S. Y., Glaser K. B., Ulevitch R. J. Lipopolysaccharide induces hyporesponsiveness to its own action in RAW 264.7 cells. J Biol Chem. 1989 Dec 25;264(36):21951–21956. [PubMed] [Google Scholar]
  28. Zhang X., Morrison D. C. Lipopolysaccharide structure-function relationship in activation versus reprogramming of mouse peritoneal macrophages. J Leukoc Biol. 1993 Nov;54(5):444–450. doi: 10.1002/jlb.54.5.444. [DOI] [PubMed] [Google Scholar]
  29. Zhang X., Morrison D. C. Lipopolysaccharide-induced selective priming effects on tumor necrosis factor alpha and nitric oxide production in mouse peritoneal macrophages. J Exp Med. 1993 Feb 1;177(2):511–516. doi: 10.1084/jem.177.2.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ziegler-Heitbrock H. W., Blumenstein M., Käfferlein E., Kieper D., Petersmann I., Endres S., Flegel W. A., Northoff H., Riethmüller G., Haas J. G. In vitro desensitization to lipopolysaccharide suppresses tumour necrosis factor, interleukin-1 and interleukin-6 gene expression in a similar fashion. Immunology. 1992 Feb;75(2):264–268. [PMC free article] [PubMed] [Google Scholar]
  31. Ziegler-Heitbrock H. W., Wedel A., Schraut W., Ströbel M., Wendelgass P., Sternsdorf T., Bäuerle P. A., Haas J. G., Riethmüller G. Tolerance to lipopolysaccharide involves mobilization of nuclear factor kappa B with predominance of p50 homodimers. J Biol Chem. 1994 Jun 24;269(25):17001–17004. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES