Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 2003 Jul 15;373(Pt 2):547–558. doi: 10.1042/BJ20030207

Fractalkine (CX3CL1) stimulated by nuclear factor kappaB (NF-kappaB)-dependent inflammatory signals induces aortic smooth muscle cell proliferation through an autocrine pathway.

Bysani Chandrasekar 1, Srinivas Mummidi 1, Rao P Perla 1, Sailaja Bysani 1, Nickolai O Dulin 1, Feng Liu 1, Peter C Melby 1
PMCID: PMC1223517  PMID: 12729461

Abstract

Fractalkine (also known as CX3CL1), a CX3C chemokine, activates and attracts monocytes/macrophages to the site of injury/inflammation. It binds to CX3C receptor 1 (CX3CR1), a pertussis toxin-sensitive G-protein-coupled receptor. In smooth muscle cells (SMCs), fractalkine is induced by proinflammatory cytokines [tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma)], which may mediate monocyte adhesion to SMCs. However, the mechanisms underlying its induction are unknown. In addition, it is unlear whether SMCs express CX3CR1. TNF-alpha activated nuclear factor kappaB (NF-kappaB) and induced fractalkine and CX3CR1 expression in a time-dependent manner in rat aortic SMCs. Transient transfections with dominant-negative (dn) inhibitory kappaB (IkappaB)-alpha, dnIkappaB-beta, dnIkappaB kinase (IKK)-gamma, kinase-dead (kd) NF-kappaB-inducing kinase (NIK) and kdIKK-beta, or pretreatment with wortmannin, Akt inhibitor, pyrrolidinecarbodithioc acid ammonium salt ('PDTC') or MG-132, significantly attenuated TNF-alpha-induced fractalkine and CX3CR1 expression. Furthermore, expression of dn TNF-alpha-receptor-associated factor 2 (TRAF2), but not dnTRAF6, inhibited TNF-alpha signal transduction. Pretreatment with pertussis toxin or neutralizing anti-CX3CR1 antibodies attenuated TNF-alpha-induced fractalkine expression, indicating that fractalkine autoregulation plays a role in TNF-alpha-induced sustained fractalkine expression. Fractalkine induced its own expression, via pertussis toxin-sensitive G-proteins, phosphoinositide 3-kinase (PI 3-kinase), phosphoinositide-dependent kinase 1 (PDK1), Akt, NIK, IKK and NF-kappaB activation, and induced SMC cell-cell adhesion and cellular proliferation. Taken together, our results demonstrate that TNF-alpha induces the expression of fractalkine and CX3CR1 in rat aortic SMCs and that this induction is mediated by NF-kappaB activation. We also show that fractalkine induces its own expression, which is mediated by the PI 3-kinase/PDK1/Akt/NIK/IKK/NF-kappaB signalling pathway. More importantly, fractalkine increased cell-cell adhesion and aortic SMC proliferation, indicating a role in initiation and progression of atherosclerotic vascular disease.

Full Text

The Full Text of this article is available as a PDF (472.2 KB).

Selected References

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

  1. Baud V., Karin M. Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol. 2001 Sep;11(9):372–377. doi: 10.1016/s0962-8924(01)02064-5. [DOI] [PubMed] [Google Scholar]
  2. Bazan J. F., Bacon K. B., Hardiman G., Wang W., Soo K., Rossi D., Greaves D. R., Zlotnik A., Schall T. J. A new class of membrane-bound chemokine with a CX3C motif. Nature. 1997 Feb 13;385(6617):640–644. doi: 10.1038/385640a0. [DOI] [PubMed] [Google Scholar]
  3. Braut-Boucher F., Pichon J., Rat P., Adolphe M., Aubery M., Font J. A non-isotopic, highly sensitive, fluorimetric, cell-cell adhesion microplate assay using calcein AM-labeled lymphocytes. J Immunol Methods. 1995 Jan 13;178(1):41–51. doi: 10.1016/0022-1759(94)00239-s. [DOI] [PubMed] [Google Scholar]
  4. Cadwallader Karen A., Condliffe Alison M., McGregor Alex, Walker Trevor R., White Jessica F., Stephens Len R., Chilvers Edwin R. Regulation of phosphatidylinositol 3-kinase activity and phosphatidylinositol 3,4,5-trisphosphate accumulation by neutrophil priming agents. J Immunol. 2002 Sep 15;169(6):3336–3344. doi: 10.4049/jimmunol.169.6.3336. [DOI] [PubMed] [Google Scholar]
  5. Cao Z., Tanaka M., Regnier C., Rothe M., Yamit-hezi A., Woronicz J. D., Fuentes M. E., Durnin M. H., Dalrymple S. A., Goeddel D. V. NF-kappa B activation by tumor necrosis factor and interleukin-1. Cold Spring Harb Symp Quant Biol. 1999;64:473–483. doi: 10.1101/sqb.1999.64.473. [DOI] [PubMed] [Google Scholar]
  6. Chandrasekar B., Mitchell D. H., Colston J. T., Freeman G. L. Regulation of CCAAT/Enhancer binding protein, interleukin-6, interleukin-6 receptor, and gp130 expression during myocardial ischemia/reperfusion. Circulation. 1999 Jan 26;99(3):427–433. doi: 10.1161/01.cir.99.3.427. [DOI] [PubMed] [Google Scholar]
  7. Chandrasekar B., Smith J. B., Freeman G. L. Ischemia-reperfusion of rat myocardium activates nuclear factor-KappaB and induces neutrophil infiltration via lipopolysaccharide-induced CXC chemokine. Circulation. 2001 May 8;103(18):2296–2302. doi: 10.1161/01.cir.103.18.2296. [DOI] [PubMed] [Google Scholar]
  8. Chandrasekar Bysani, Colston James T., de la Rosa Sam D., Rao Perla P., Freeman Gregory L. TNF-alpha and H2O2 induce IL-18 and IL-18R beta expression in cardiomyocytes via NF-kappa B activation. Biochem Biophys Res Commun. 2003 Apr 18;303(4):1152–1158. doi: 10.1016/s0006-291x(03)00496-0. [DOI] [PubMed] [Google Scholar]
  9. Chandrasekar Bysani, Melby Peter C., Sarau Henry M., Raveendran Muthuswamy, Perla Rao P., Marelli-Berg Federica M., Dulin Nickolai O., Singh Ishwar S. Chemokine-cytokine cross-talk. The ELR+ CXC chemokine LIX (CXCL5) amplifies a proinflammatory cytokine response via a phosphatidylinositol 3-kinase-NF-kappa B pathway. J Biol Chem. 2002 Dec 4;278(7):4675–4686. doi: 10.1074/jbc.M207006200. [DOI] [PubMed] [Google Scholar]
  10. Choe N., Zhang J., Iwagaki A., Tanaka S., Hemenway D. R., Kagan E. Asbestos exposure upregulates the adhesion of pleural leukocytes to pleural mesothelial cells via VCAM-1. Am J Physiol. 1999 Aug;277(2 Pt 1):L292–L300. doi: 10.1152/ajplung.1999.277.2.L292. [DOI] [PubMed] [Google Scholar]
  11. Corda S., Laplace C., Vicaut E., Duranteau J. Rapid reactive oxygen species production by mitochondria in endothelial cells exposed to tumor necrosis factor-alpha is mediated by ceramide. Am J Respir Cell Mol Biol. 2001 Jun;24(6):762–768. doi: 10.1165/ajrcmb.24.6.4228. [DOI] [PubMed] [Google Scholar]
  12. Datta S. R., Brunet A., Greenberg M. E. Cellular survival: a play in three Akts. Genes Dev. 1999 Nov 15;13(22):2905–2927. doi: 10.1101/gad.13.22.2905. [DOI] [PubMed] [Google Scholar]
  13. Druey K. M., Blumer K. J., Kang V. H., Kehrl J. H. Inhibition of G-protein-mediated MAP kinase activation by a new mammalian gene family. Nature. 1996 Feb 22;379(6567):742–746. doi: 10.1038/379742a0. [DOI] [PubMed] [Google Scholar]
  14. Dulin N. O., Sorokin A., Reed E., Elliott S., Kehrl J. H., Dunn M. J. RGS3 inhibits G protein-mediated signaling via translocation to the membrane and binding to Galpha11. Mol Cell Biol. 1999 Jan;19(1):714–723. doi: 10.1128/mcb.19.1.714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Faure S., Meyer L., Costagliola D., Vaneensberghe C., Genin E., Autran B., Delfraissy J. F., McDermott D. H., Murphy P. M., Debré P. Rapid progression to AIDS in HIV+ individuals with a structural variant of the chemokine receptor CX3CR1. Science. 2000 Mar 24;287(5461):2274–2277. doi: 10.1126/science.287.5461.2274. [DOI] [PubMed] [Google Scholar]
  16. Fong A. M., Robinson L. A., Steeber D. A., Tedder T. F., Yoshie O., Imai T., Patel D. D. Fractalkine and CX3CR1 mediate a novel mechanism of leukocyte capture, firm adhesion, and activation under physiologic flow. J Exp Med. 1998 Oct 19;188(8):1413–1419. doi: 10.1084/jem.188.8.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fong A. M., Robinson L. A., Steeber D. A., Tedder T. F., Yoshie O., Imai T., Patel D. D. Fractalkine and CX3CR1 mediate a novel mechanism of leukocyte capture, firm adhesion, and activation under physiologic flow. J Exp Med. 1998 Oct 19;188(8):1413–1419. doi: 10.1084/jem.188.8.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Foukas Lazaros C., Daniele Nathalie, Ktori Chariklia, Anderson Karen E., Jensen Jorgen, Shepherd Peter R. Direct effects of caffeine and theophylline on p110 delta and other phosphoinositide 3-kinases. Differential effects on lipid kinase and protein kinase activities. J Biol Chem. 2002 Jul 26;277(40):37124–37130. doi: 10.1074/jbc.M202101200. [DOI] [PubMed] [Google Scholar]
  19. Garcia G. E., Xia Y., Chen S., Wang Y., Ye R. D., Harrison J. K., Bacon K. B., Zerwes H. G., Feng L. NF-kappaB-dependent fractalkine induction in rat aortic endothelial cells stimulated by IL-1beta, TNF-alpha, and LPS. J Leukoc Biol. 2000 Apr;67(4):577–584. doi: 10.1002/jlb.67.4.577. [DOI] [PubMed] [Google Scholar]
  20. Garton K. J., Gough P. J., Blobel C. P., Murphy G., Greaves D. R., Dempsey P. J., Raines E. W. Tumor necrosis factor-alpha-converting enzyme (ADAM17) mediates the cleavage and shedding of fractalkine (CX3CL1). J Biol Chem. 2001 Aug 8;276(41):37993–38001. doi: 10.1074/jbc.M106434200. [DOI] [PubMed] [Google Scholar]
  21. Han Y., He T., Huang D. R., Pardo C. A., Ransohoff R. M. TNF-alpha mediates SDF-1 alpha-induced NF-kappa B activation and cytotoxic effects in primary astrocytes. J Clin Invest. 2001 Aug;108(3):425–435. doi: 10.1172/JCI12629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Harrison J. K., Jiang Y., Wees E. A., Salafranca M. N., Liang H. X., Feng L., Belardinelli L. Inflammatory agents regulate in vivo expression of fractalkine in endothelial cells of the rat heart. J Leukoc Biol. 1999 Dec;66(6):937–944. doi: 10.1002/jlb.66.6.937. [DOI] [PubMed] [Google Scholar]
  23. Hu Y., Qiao L., Wang S., Rong S. B., Meuillet E. J., Berggren M., Gallegos A., Powis G., Kozikowski A. P. 3-(Hydroxymethyl)-bearing phosphatidylinositol ether lipid analogues and carbonate surrogates block PI3-K, Akt, and cancer cell growth. J Med Chem. 2000 Aug 10;43(16):3045–3051. doi: 10.1021/jm000117y. [DOI] [PubMed] [Google Scholar]
  24. Idriss H. T., Naismith J. H. TNF alpha and the TNF receptor superfamily: structure-function relationship(s). Microsc Res Tech. 2000 Aug 1;50(3):184–195. doi: 10.1002/1097-0029(20000801)50:3<184::AID-JEMT2>3.0.CO;2-H. [DOI] [PubMed] [Google Scholar]
  25. Imai T., Hieshima K., Haskell C., Baba M., Nagira M., Nishimura M., Kakizaki M., Takagi S., Nomiyama H., Schall T. J. Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both leukocyte migration and adhesion. Cell. 1997 Nov 14;91(4):521–530. doi: 10.1016/s0092-8674(00)80438-9. [DOI] [PubMed] [Google Scholar]
  26. Inngjerdingen M., Damaj B., Maghazachi A. A. Expression and regulation of chemokine receptors in human natural killer cells. Blood. 2001 Jan 15;97(2):367–375. doi: 10.1182/blood.v97.2.367. [DOI] [PubMed] [Google Scholar]
  27. Libby Peter. Inflammation in atherosclerosis. Nature. 2002 Dec 19;420(6917):868–874. doi: 10.1038/nature01323. [DOI] [PubMed] [Google Scholar]
  28. Ludwig Andreas, Berkhout Theo, Moores Kitty, Groot Pieter, Chapman Gayle. Fractalkine is expressed by smooth muscle cells in response to IFN-gamma and TNF-alpha and is modulated by metalloproteinase activity. J Immunol. 2002 Jan 15;168(2):604–612. doi: 10.4049/jimmunol.168.2.604. [DOI] [PubMed] [Google Scholar]
  29. Mayne M., Ni W., McKenna R., Power C. Antisense oligodeoxynucleotides targeting internal exon sequences efficiently regulate TNF-alpha expression. Antisense Nucleic Acid Drug Dev. 1999 Apr;9(2):135–144. doi: 10.1089/oli.1.1999.9.135. [DOI] [PubMed] [Google Scholar]
  30. McDermott D. H., Halcox J. P., Schenke W. H., Waclawiw M. A., Merrell M. N., Epstein N., Quyyumi A. A., Murphy P. M. Association between polymorphism in the chemokine receptor CX3CR1 and coronary vascular endothelial dysfunction and atherosclerosis. Circ Res. 2001 Aug 31;89(5):401–407. doi: 10.1161/hh1701.095642. [DOI] [PubMed] [Google Scholar]
  31. Meng Fanyin, Liu Li, Chin Paul C., D'Mello Santosh R. Akt is a downstream target of NF-kappa B. J Biol Chem. 2002 Jun 6;277(33):29674–29680. doi: 10.1074/jbc.M112464200. [DOI] [PubMed] [Google Scholar]
  32. Meucci O., Fatatis A., Simen A. A., Miller R. J. Expression of CX3CR1 chemokine receptors on neurons and their role in neuronal survival. Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8075–8080. doi: 10.1073/pnas.090017497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Moatti D., Faure S., Fumeron F., Amara M. el-W, Seknadji P., McDermott D. H., Debré P., Aumont M. C., Murphy P. M., de Prost D. Polymorphism in the fractalkine receptor CX3CR1 as a genetic risk factor for coronary artery disease. Blood. 2001 Apr 1;97(7):1925–1928. doi: 10.1182/blood.v97.7.1925. [DOI] [PubMed] [Google Scholar]
  34. Murray D. R., Prabhu S. D., Chandrasekar B. Chronic beta-adrenergic stimulation induces myocardial proinflammatory cytokine expression. Circulation. 2000 May 23;101(20):2338–2341. doi: 10.1161/01.cir.101.20.2338. [DOI] [PubMed] [Google Scholar]
  35. Niu Jiaxin, Scheschonka Astrid, Druey Kirk M., Davis Amanda, Reed Eleanor, Kolenko Vladimir, Bodnar Richard, Voyno-Yasenetskaya Tatyana, Du Xiaoping, Kehrl John. RGS3 interacts with 14-3-3 via the N-terminal region distinct from the RGS (regulator of G-protein signalling) domain. Biochem J. 2002 Aug 1;365(Pt 3):677–684. doi: 10.1042/BJ20020390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Onuffer James J., Horuk Richard. Chemokines, chemokine receptors and small-molecule antagonists: recent developments. Trends Pharmacol Sci. 2002 Oct;23(10):459–467. doi: 10.1016/s0165-6147(02)02064-3. [DOI] [PubMed] [Google Scholar]
  37. Orlov S. N., Tremblay J., Hamet P. cAMP signaling inhibits dihydropyridine-sensitive Ca2+ influx in vascular smooth muscle cells. Hypertension. 1996 Mar;27(3 Pt 2):774–780. doi: 10.1161/01.hyp.27.3.774. [DOI] [PubMed] [Google Scholar]
  38. Ozes O. N., Mayo L. D., Gustin J. A., Pfeffer S. R., Pfeffer L. M., Donner D. B. NF-kappaB activation by tumour necrosis factor requires the Akt serine-threonine kinase. Nature. 1999 Sep 2;401(6748):82–85. doi: 10.1038/43466. [DOI] [PubMed] [Google Scholar]
  39. Pollman M. J., Yamada T., Horiuchi M., Gibbons G. H. Vasoactive substances regulate vascular smooth muscle cell apoptosis. Countervailing influences of nitric oxide and angiotensin II. Circ Res. 1996 Oct;79(4):748–756. doi: 10.1161/01.res.79.4.748. [DOI] [PubMed] [Google Scholar]
  40. Rossi D., Zlotnik A. The biology of chemokines and their receptors. Annu Rev Immunol. 2000;18:217–242. doi: 10.1146/annurev.immunol.18.1.217. [DOI] [PubMed] [Google Scholar]
  41. Schreck R., Meier B., Männel D. N., Dröge W., Baeuerle P. A. Dithiocarbamates as potent inhibitors of nuclear factor kappa B activation in intact cells. J Exp Med. 1992 May 1;175(5):1181–1194. doi: 10.1084/jem.175.5.1181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tartaglia L. A., Goeddel D. V. Two TNF receptors. Immunol Today. 1992 May;13(5):151–153. doi: 10.1016/0167-5699(92)90116-O. [DOI] [PubMed] [Google Scholar]
  43. Terkeltaub R., Boisvert W. A., Curtiss L. K. Chemokines and atherosclerosis. Curr Opin Lipidol. 1998 Oct;9(5):397–405. doi: 10.1097/00041433-199810000-00003. [DOI] [PubMed] [Google Scholar]
  44. Toker A., Cantley L. C. Signalling through the lipid products of phosphoinositide-3-OH kinase. Nature. 1997 Jun 12;387(6634):673–676. doi: 10.1038/42648. [DOI] [PubMed] [Google Scholar]
  45. Umehara H., Bloom E., Okazaki T., Domae N., Imai T. Fractalkine and vascular injury. Trends Immunol. 2001 Nov;22(11):602–607. doi: 10.1016/s1471-4906(01)02051-8. [DOI] [PubMed] [Google Scholar]
  46. Wick M. J., Dong L. Q., Riojas R. A., Ramos F. J., Liu F. Mechanism of phosphorylation of protein kinase B/Akt by a constitutively active 3-phosphoinositide-dependent protein kinase-1. J Biol Chem. 2000 Dec 22;275(51):40400–40406. doi: 10.1074/jbc.M003937200. [DOI] [PubMed] [Google Scholar]
  47. Wong Brian W. C., Wong Donald, McManus Bruce M. Characterization of fractalkine (CX3CL1) and CX3CR1 in human coronary arteries with native atherosclerosis, diabetes mellitus, and transplant vascular disease. Cardiovasc Pathol. 2002 Nov-Dec;11(6):332–338. doi: 10.1016/s1054-8807(02)00111-4. [DOI] [PubMed] [Google Scholar]
  48. Zha J., Harada H., Osipov K., Jockel J., Waksman G., Korsmeyer S. J. BH3 domain of BAD is required for heterodimerization with BCL-XL and pro-apoptotic activity. J Biol Chem. 1997 Sep 26;272(39):24101–24104. doi: 10.1074/jbc.272.39.24101. [DOI] [PubMed] [Google Scholar]
  49. Zha J., Harada H., Yang E., Jockel J., Korsmeyer S. J. Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L) Cell. 1996 Nov 15;87(4):619–628. doi: 10.1016/s0092-8674(00)81382-3. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES