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. 2005 Jan;64(1):21–28. doi: 10.1136/ard.2003.018705

Up regulated expression of fractalkine/CX3CL1 and CX3CR1 in patients with systemic sclerosis

M Hasegawa 1, S Sato 1, T Echigo 1, Y Hamaguchi 1, M Yasui 1, K Takehara 1
PMCID: PMC1755178  PMID: 15608300

Abstract

Background: Fractalkine expressed on endothelial cells mediates activation and adhesion of leucocytes expressing its receptor, CX3CR1. Soluble fractalkine exhibits chemotactic activity for leucocytes expressing CX3CR1.

Objective: To determine the role of fractalkine and its receptor in systemic sclerosis (SSc) by assessing their expression levels in patients with this disease.

Methods: The expression of fractalkine and CX3CR1 in the skin and lung tissues was immunohistochemically examined. Circulating soluble fractalkine levels were examined by enzyme linked immunosorbent assay (ELISA). Blood samples from patients with SSc were stained for CX3CR1 with flow cytometric analysis.

Results: CX3CR1 levels on peripheral monocytes/macrophages and T cells were found to be raised in patients with diffuse cutaneous SSc. The numbers of cells expressing CX3CR1, including monocytes/macrophages, were increased in the lesional skin and lung tissues from patients with diffuse cutaneous SSc. Fractalkine was strongly expressed on endothelial cells in the affected skin and lung tissues. Soluble fractalkine levels were significantly raised in sera and were associated with raised erythrocyte sedimentation rates, digital ischaemia, and severity of pulmonary fibrosis.

Conclusions: Up regulated expression of fractalkine and CX3CR1 cooperatively augments the recruitment of mononuclear cells expressing CX3CR1 into the affected tissue of SSc, leading to inflammation and vascular injury.

Full Text

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Figure 1.

Figure 1

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 (A) CX3CR1 expression in normal skin tissues (CTL) and lesional skin tissues from patients with dSSc with rapidly progressing skin thickening. (B) CX3CR1 expression in normal lung tissues and affected lung tissues from patients with dSSc with progressive pulmonary fibrosis. Staining with control polyclonal rabbit IgG in affected skin and lung sections from SSc was shown as a negative control. Magnification x250.

Figure 2.

Figure 2

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 Representative expression of CX3CR1 on PBMC from patients with dSSc and normal controls. All samples were stained in parallel by two colour immunofluorescence staining and analysed sequentially by flow cytometry with identical instrument setting. Relative fluorescence intensity is shown on a four decade log scale. The percentage represents the frequency of each leucocyte subset among total mononuclear cells.

Figure 3.

Figure 3

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 Expression of CX3CR1 on leucocyte subpopulations from patients with lSSc, patients with dSSc, and normal controls. CX3CR1 levels were assessed by two colour immunofluorescence staining with flow cytometric analysis. All samples were stained and analysed sequentially by flow cytometry in parallel with the same instrument settings.

Figure 4.

Figure 4

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 (A) FKN expression in normal skin tissues and skin tissues from patients with dSSc with rapidly progressing skin thickening. (B) FKN expression in normal lung tissues and affected lung tissues from patients with dSSc with progressive pulmonary fibrosis. Staining with control polyclonal goat IgG in affected skin and lung sections from SSc was shown as a negative control. Magnification x400.

Figure 5.

Figure 5

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 (A) Serum levels of sFKN in patients with patients with lSSc, patients with dSSc, and normal controls (CTL). (B) Serum levels of sFKN in patients with SSc with (PF+) and without (PF–) pulmonary fibrosis. Soluble FKN levels were determined using a specific ELISA. The short bar indicates the mean value in each group. The broken line indicates the cut off value (mean + 3SD of the CTL samples).

Figure 6.

Figure 6

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 Correlations of serum sFKN levels with laboratory data in patients with SSc. The correlation of sFKN levels with (A) %VC and (B) TLCO is shown. Soluble FKN levels were determined using a specific ELISA.

Selected References

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

  1. Ancuta Petronela, Rao Ravi, Moses Ashlee, Mehle Andrew, Shaw Sunil K., Luscinskas F. William, Gabuzda Dana. Fractalkine preferentially mediates arrest and migration of CD16+ monocytes. J Exp Med. 2003 Jun 16;197(12):1701–1707. doi: 10.1084/jem.20022156. [DOI] [PMC free article] [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. Clements P. J., Lachenbruch P. A., Seibold J. R., Zee B., Steen V. D., Brennan P., Silman A. J., Allegar N., Varga J., Massa M. Skin thickness score in systemic sclerosis: an assessment of interobserver variability in 3 independent studies. J Rheumatol. 1993 Nov;20(11):1892–1896. [PubMed] [Google Scholar]
  4. Combadière Christophe, Potteaux Stéphane, Gao Ji-Liang, Esposito Bruno, Casanova Saveria, Lee Eric J., Debré Patrice, Tedgui Alain, Murphy Philip M., Mallat Ziad. Decreased atherosclerotic lesion formation in CX3CR1/apolipoprotein E double knockout mice. Circulation. 2003 Feb 25;107(7):1009–1016. doi: 10.1161/01.cir.0000057548.68243.42. [DOI] [PubMed] [Google Scholar]
  5. Denton C. P., Abraham D. J. Transforming growth factor-beta and connective tissue growth factor: key cytokines in scleroderma pathogenesis. Curr Opin Rheumatol. 2001 Nov;13(6):505–511. doi: 10.1097/00002281-200111000-00010. [DOI] [PubMed] [Google Scholar]
  6. Feng L., Chen S., Garcia G. E., Xia Y., Siani M. A., Botti P., Wilson C. B., Harrison J. K., Bacon K. B. Prevention of crescentic glomerulonephritis by immunoneutralization of the fractalkine receptor CX3CR1 rapid communication. Kidney Int. 1999 Aug;56(2):612–620. doi: 10.1046/j.1523-1755.1999.00604.x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Fraticelli P., Sironi M., Bianchi G., D'Ambrosio D., Albanesi C., Stoppacciaro A., Chieppa M., Allavena P., Ruco L., Girolomoni G. Fractalkine (CX3CL1) as an amplification circuit of polarized Th1 responses. J Clin Invest. 2001 May;107(9):1173–1181. doi: 10.1172/JCI11517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Furuichi K., Wada T., Iwata Y., Sakai N., Yoshimoto K., Shimizu M., Kobayashi K., Takasawa K., Kida H., Takeda S. Upregulation of fractalkine in human crescentic glomerulonephritis. Nephron. 2001 Apr;87(4):314–320. doi: 10.1159/000045936. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Gruschwitz M., Sepp N., Kofler H., Wick G. Expression of class II-MHC antigens in the dermis of patients with progressive systemic sclerosis. Immunobiology. 1991 Jun;182(3-4):234–255. doi: 10.1016/S0171-2985(11)80660-1. [DOI] [PubMed] [Google Scholar]
  12. Gudbjörnsson B., Hällgren R., Nettelbladt O., Gustafsson R., Mattsson A., af Geijerstam E., Tötterman T. H. Phenotypic and functional activation of alveolar macrophages, T lymphocytes and NK cells in patients with systemic sclerosis and primary Sjögren's syndrome. Ann Rheum Dis. 1994 Sep;53(9):574–579. doi: 10.1136/ard.53.9.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Ishikawa O., Ishikawa H. Macrophage infiltration in the skin of patients with systemic sclerosis. J Rheumatol. 1992 Aug;19(8):1202–1206. [PubMed] [Google Scholar]
  15. Kahaleh M. B., Fan P. S. Mechanism of serum-mediated endothelial injury in scleroderma: identification of a granular enzyme in scleroderma skin and sera. Clin Immunol Immunopathol. 1997 Apr;83(1):32–40. doi: 10.1006/clin.1996.4322. [DOI] [PubMed] [Google Scholar]
  16. Kanazawa N., Nakamura T., Tashiro K., Muramatsu M., Morita K., Yoneda K., Inaba K., Imamura S., Honjo T. Fractalkine and macrophage-derived chemokine: T cell-attracting chemokines expressed in T cell area dendritic cells. Eur J Immunol. 1999 Jun;29(6):1925–1932. doi: 10.1002/(SICI)1521-4141(199906)29:06<1925::AID-IMMU1925>3.0.CO;2-U. [DOI] [PubMed] [Google Scholar]
  17. Kräling B. M., Maul G. G., Jimenez S. A. Mononuclear cellular infiltrates in clinically involved skin from patients with systemic sclerosis of recent onset predominantly consist of monocytes/macrophages. Pathobiology. 1995;63(1):48–56. doi: 10.1159/000163933. [DOI] [PubMed] [Google Scholar]
  18. LeRoy E. C., Black C., Fleischmajer R., Jablonska S., Krieg T., Medsger T. A., Jr, Rowell N., Wollheim F. Scleroderma (systemic sclerosis): classification, subsets and pathogenesis. J Rheumatol. 1988 Feb;15(2):202–205. [PubMed] [Google Scholar]
  19. Lesnik Philippe, Haskell Christopher A., Charo Israel F. Decreased atherosclerosis in CX3CR1-/- mice reveals a role for fractalkine in atherogenesis. J Clin Invest. 2003 Feb;111(3):333–340. doi: 10.1172/JCI15555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Nagaoka T., Kaburagi Y., Hamaguchi Y., Hasegawa M., Takehara K., Steeber D. A., Tedder T. F., Sato S. Delayed wound healing in the absence of intercellular adhesion molecule-1 or L-selectin expression. Am J Pathol. 2000 Jul;157(1):237–247. doi: 10.1016/S0002-9440(10)64534-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nanki Toshihiro, Imai Toshio, Nagasaka Kenji, Urasaki Yasuyo, Nonomura Yoshinori, Taniguchi Ken, Hayashida Kenji, Hasegawa Jun, Yoshie Osamu, Miyasaka Nobuyuki. Migration of CX3CR1-positive T cells producing type 1 cytokines and cytotoxic molecules into the synovium of patients with rheumatoid arthritis. Arthritis Rheum. 2002 Nov;46(11):2878–2883. doi: 10.1002/art.10622. [DOI] [PubMed] [Google Scholar]
  24. Nishimura Miyuki, Umehara Hisanori, Nakayama Takashi, Yoneda Osamu, Hieshima Kunio, Kakizaki Mayumi, Dohmae Naochika, Yoshie Osamu, Imai Toshio. Dual functions of fractalkine/CX3C ligand 1 in trafficking of perforin+/granzyme B+ cytotoxic effector lymphocytes that are defined by CX3CR1 expression. J Immunol. 2002 Jun 15;168(12):6173–6180. doi: 10.4049/jimmunol.168.12.6173. [DOI] [PubMed] [Google Scholar]
  25. Pan Y., Lloyd C., Zhou H., Dolich S., Deeds J., Gonzalo J. A., Vath J., Gosselin M., Ma J., Dussault B. Neurotactin, a membrane-anchored chemokine upregulated in brain inflammation. Nature. 1997 Jun 5;387(6633):611–617. doi: 10.1038/42491. [DOI] [PubMed] [Google Scholar]
  26. Prescott R. J., Freemont A. J., Jones C. J., Hoyland J., Fielding P. Sequential dermal microvascular and perivascular changes in the development of scleroderma. J Pathol. 1992 Mar;166(3):255–263. doi: 10.1002/path.1711660307. [DOI] [PubMed] [Google Scholar]
  27. Roumm A. D., Whiteside T. L., Medsger T. A., Jr, Rodnan G. P. Lymphocytes in the skin of patients with progressive systemic sclerosis. Quantification, subtyping, and clinical correlations. Arthritis Rheum. 1984 Jun;27(6):645–653. doi: 10.1002/art.1780270607. [DOI] [PubMed] [Google Scholar]
  28. Ruth J. H., Rottman J. B., Katschke K. J., Jr, Qin S., Wu L., LaRosa G., Ponath P., Pope R. M., Koch A. E. Selective lymphocyte chemokine receptor expression in the rheumatoid joint. Arthritis Rheum. 2001 Dec;44(12):2750–2760. doi: 10.1002/1529-0131(200112)44:12<2750::aid-art462>3.0.co;2-c. [DOI] [PubMed] [Google Scholar]
  29. Sato S. Abnormalities of adhesion molecules and chemokines in scleroderma. Curr Opin Rheumatol. 1999 Nov;11(6):503–507. [PubMed] [Google Scholar]
  30. Sato S., Ihn H., Kikuchi K., Takehara K. Antihistone antibodies in systemic sclerosis. Association with pulmonary fibrosis. Arthritis Rheum. 1994 Mar;37(3):391–394. doi: 10.1002/art.1780370313. [DOI] [PubMed] [Google Scholar]
  31. Sato S., Miller A. S., Inaoki M., Bock C. B., Jansen P. J., Tang M. L., Tedder T. F. CD22 is both a positive and negative regulator of B lymphocyte antigen receptor signal transduction: altered signaling in CD22-deficient mice. Immunity. 1996 Dec;5(6):551–562. doi: 10.1016/s1074-7613(00)80270-8. [DOI] [PubMed] [Google Scholar]
  32. Scharffetter K., Lankat-Buttgereit B., Krieg T. Localization of collagen mRNA in normal and scleroderma skin by in-situ hybridization. Eur J Clin Invest. 1988 Feb;18(1):9–17. doi: 10.1111/j.1365-2362.1988.tb01158.x. [DOI] [PubMed] [Google Scholar]
  33. Silver R. M., Miller K. S., Kinsella M. B., Smith E. A., Schabel S. I. Evaluation and management of scleroderma lung disease using bronchoalveolar lavage. Am J Med. 1990 May;88(5):470–476. doi: 10.1016/0002-9343(90)90425-d. [DOI] [PubMed] [Google Scholar]
  34. Steen V. D., Powell D. L., Medsger T. A., Jr Clinical correlations and prognosis based on serum autoantibodies in patients with systemic sclerosis. Arthritis Rheum. 1988 Feb;31(2):196–203. doi: 10.1002/art.1780310207. [DOI] [PubMed] [Google Scholar]
  35. Takehara Kazuhiko. Hypothesis: pathogenesis of systemic sclerosis. J Rheumatol. 2003 Apr;30(4):755–759. [PubMed] [Google Scholar]
  36. Tsou C. L., Haskell C. A., Charo I. F. Tumor necrosis factor-alpha-converting enzyme mediates the inducible cleavage of fractalkine. J Biol Chem. 2001 Sep 24;276(48):44622–44626. doi: 10.1074/jbc.M107327200. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Umehara H., Goda S., Imai T., Nagano Y., Minami Y., Tanaka Y., Okazaki T., Bloom E. T., Domae N. Fractalkine, a CX3C-chemokine, functions predominantly as an adhesion molecule in monocytic cell line THP-1. Immunol Cell Biol. 2001 Jun;79(3):298–302. doi: 10.1046/j.1440-1711.2001.01004.x. [DOI] [PubMed] [Google Scholar]
  39. Volin M. V., Woods J. M., Amin M. A., Connors M. A., Harlow L. A., Koch A. E. Fractalkine: a novel angiogenic chemokine in rheumatoid arthritis. Am J Pathol. 2001 Oct;159(4):1521–1530. doi: 10.1016/S0002-9440(10)62537-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yoneda O., Imai T., Goda S., Inoue H., Yamauchi A., Okazaki T., Imai H., Yoshie O., Bloom E. T., Domae N. Fractalkine-mediated endothelial cell injury by NK cells. J Immunol. 2000 Apr 15;164(8):4055–4062. doi: 10.4049/jimmunol.164.8.4055. [DOI] [PubMed] [Google Scholar]

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