Advertisement

Journal of Clinical Immunology

, Volume 15, Issue 6, pp 300–311 | Cite as

Expression of CD44 variants in human inflammatory synovitis

  • Laura P. Hale
  • Barton F. Haynes
  • S. Spence McCachren
Original Articles

Abstract

The cell surface hyaluronate receptor CD44 has previously been shown to have immunomodulatory activity and to be upregulated in inflammatory synovitis. Since these findings were reported, the genomic structure of CD44 has been delineated, and multiple splice variants have been described. Therefore, we determined which CD44 variant exons are present during inflammatory synovitis by a combination of northern blot analysis and reverse transcription followed by polymerase chain reaction amplification of synovial RNA. Immunohistochemical staining was used to define the sites of expression of individual v6 and v9 exons in synovial tissue. The standard (S) or hematopoietic isoform, CD44S, was the predominant form of CD44 expressed in synovium and was expressed by most cell types. Other isoforms, containing alternatively spliced exons in the proximal extracellular domain, were found by RT-PCR, but at lower levels than CD44S. The second most prevalent form was CD44E, which has an insertion of three exons (v8-v10) in the proximal extracellular domain. Immunohistochemical studies showed that reactivity with v9-specific antibodies was primarily in macrophages, particularly those in the synovial lining layer. CD44 exon v6, previously reported to be important in immune activation and in epithelial tumor metastasis, was also expressed in synovial lining cells and in occasional synovial interstitial cells. The presence of CD44 variants containing v9 in rheumatoid synovial macrophages may be important in the adhesion and activation of mononuclear phagocytes in the synovium and, thus, may be a target for novel antiinflammatory therapies in the future. The role of CD44 isoforms in cellular adhesion, immune activation, and joint erosion in inflammatory synovitis deserves further study.

Key words

CD44 isoforms arthritis inflammation 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Springer TA: Adhesion receptors of the immune system. Nature 346:425–434, 1990PubMedGoogle Scholar
  2. 2.
    Haynes BF, Hale LP, Denning SM, Le PT, Singer KH: The role of leukocyte adhesion molecules in cellular interaction: Implications for the pathogenesis of inflammatory synovitis. Springer Semin Immunopathol 11:163–185, 1989PubMedGoogle Scholar
  3. 3.
    Hemler ME: Adhesive protein receptors and hematopoietic cells. Immunology Today 9:109–113, 1988PubMedGoogle Scholar
  4. 4.
    Cronstein BN, Weissmann G: The adhesion molecules of inflammation. Arth Rheum 36:147–157, 1993Google Scholar
  5. 5.
    Denning SM, Le PT, Singer KH, Haynes BF: Antibodies against the CD44 p80, lymphocyte homing receptor molecule augment human peripheral blood T cell activation. J Immunol 144:7–15, 1990PubMedGoogle Scholar
  6. 6.
    Hale LP, Singer KH, Haynes BF: CD44 antibody against In(Lu)related p80, lymphocyte-homing receptor molecule inhibits the binding of human erythrocytes to T cells. J Immunol 143:3944–3948, 1989PubMedGoogle Scholar
  7. 7.
    Koopman G, van Kooyk Y, de Graaff M, Meyer CJ, Figdor CG, Pals ST: Triggering of the CD44 antigen on T lymphocytes promotes T cell adhesion through the LFA-1 pathway. J Immunol 145:3589–3593, 1990PubMedGoogle Scholar
  8. 8.
    Webb DSA, Shimuzu Y, Van Seventer GA, Shaw S, Gerrard TL: LFA-3, CD44 and Cd45: physiologic triggers of human monocyte TNF and IL-1 release. Science 249:1295–1297, 1990PubMedGoogle Scholar
  9. 9.
    Jalkanen S, Steere AC, Fox RA, Butcher EC: A distinct endothelial cell recognition system that controls lymphocyte traffic into inflamed synovium. Science 233:556–558, 1986PubMedGoogle Scholar
  10. 10.
    Jalkanen S, Bargatze RF, de los Toyos J, Butcher EC: Lymphocyte recognition of high endothelium: antibodies to distinct epitopes of an 85–95kd glycoprotein antigen differentially inhibit lymphocyte binding to lymph node, mucosal or synovial endothelial cells. J Cell Biol 105:983–990, 1987PubMedGoogle Scholar
  11. 11.
    Haynes BF, Hale LP, Patton KL, Martin ME, McCallum RM: Measurement of an adhesion molecule as an indicator of inflammatory disease activity. Up-regulation of the receptor for hyaluronate (CD44) in rheumatoid arthritis. Arth Rheum 34:1434–1443, 1991Google Scholar
  12. 12.
    Johnson BA, Haines GK, Harlow LA, Koch AE: Adhesion molecule expression in human synovial tissue. Arth Rheum 36: 137–146, 1993Google Scholar
  13. 13.
    Jalkanen S, Jalkanen M, Bargatze R, Tammi M, Butcher EC: Biochemical properties of glycoproteins involved in lymphocyte recognition of high endothelial venules in man. J Immunol 141: 1615–1623, 1988PubMedGoogle Scholar
  14. 14.
    Gunthert U, Hofmann M, Rudy W, Reber S, Zoller M, Haussmann I, Matzku S, Wenzel A, Ponta H, Herrlich P: A new variant of glycoprotein CD44 confers metastatic potential to rat carcinoma cells. Cell 65:13–24, 1991PubMedGoogle Scholar
  15. 15.
    Screaton GR, Bell MV, Jackson DG, Cornelis FB, Gerth U, Bell JI: Genomic structure of DNA encoding the lymphocyte homing receptor CD44 reveals at least 12 alternatively spliced exons. Proc Natl Acad Sci USA 89:12160–12164, 1992PubMedGoogle Scholar
  16. 16.
    Arch R, Wirth K, Hofmann M, Ponta H, Matzku S, Herrlich P, Zoller M: Participation in normal immune responses of a metastasis-inducing splice variant of CD44. Science 257:682–685, 1992PubMedGoogle Scholar
  17. 17.
    Koopman G, Heider K-H, Horst E, Adolf GR, van den Berg F, Ponta H, Herrlich P, Pals ST: Activated human lymphocytes and aggressive non-Hodgkins lymphomas express a homologue of the rat metastasis-associated variant of CD44. J Exp Med 177:897–904, 1993PubMedGoogle Scholar
  18. 18.
    Mackay CR, Terpe HJ, Stauder R, Marston WL, Stark H, Gunthert U: Expression and modulation of CD44 variant isoforms in humans. J Cell Biol 124:71–82, 1994PubMedGoogle Scholar
  19. 19.
    Hale LP, Patel DD, Liao H-X, McCachren SS, Haynes BF: Reactivity of the CD44 workshop panel with multiple hematopoietic and nonhematopoietic cell lines. Tissue Antigens 42:266, 1993 (abstr)Google Scholar
  20. 20.
    Heider K-H, Hofmann M, Horst E, van den Berg F, Ponta H, Herrlich P, Pals ST: A human homologue of the rat metastasisassociated variant of CD44 is expressed in colorectal carcinomas and adenomatous polyps. J Cell Biol 120:227–233, 1993PubMedGoogle Scholar
  21. 21.
    Rudy W, Hofmann M, Schwartz-Albiez R, Zoller M, Heider KH, Ponta H, Herrlich P: The two major CD44 proteins expressed on a metastatic rat tumor cell line are derived from different splice variants: Each one individually suffices to confer metastatic behavior. Cancer Res 53:1262–1268, 1993PubMedGoogle Scholar
  22. 22.
    Matsumura Y, Tarin D: Significance of CD44 gene products for cancer diagnosis and disease evaluation. Lancet 340:1053–1058, 1992PubMedGoogle Scholar
  23. 23.
    Denning SM: CD44 antibodies enhance CD2-mediated T cell activation. Tissue Antigens 42:263, 1993 (abstr)Google Scholar
  24. 24.
    McCachren SS, Greer PK, Niedel JE: Regulation of human synovial fibroblast collagenase messenger RNA by interleukin-1. Arth Rheum 32:1539–1545, 1989Google Scholar
  25. 25.
    McDonald JA, Broekelmann TJ, Matheke ML, Crouch E, Koo M, Kuhn C: A monoclonal antibody to the carboxyterminal domain of procollagen type I visualizes collagen-synthesizing fibroblasts. J Clin Invest 78:1237–1244, 1986PubMedGoogle Scholar
  26. 26.
    Weinberg JB, Athens JW: The mononuclear phagocyte system. In Wintrobe's Clinical Hematology, 9th ed. Philadelphia, Lea and Febiger, 1993, pp 267–298Google Scholar
  27. 27.
    James-Yarish M, Bradley WG, Emmanuel PJ, Good RA, Day NK: Detection of cell specific cluster determinant expression by reverse transcriptase polymerase chain reaction. J Immunol Methods 169:73–82, 1994PubMedGoogle Scholar
  28. 28.
    Uphoff CC, Hu Z-B, Gignac SM, Ma W, Rainey FA, Kreutz M, Ludwig W-D, Drexler HG: Characterization of the monocytespecific esterase (MSE) gene. Leukemia 8:1510–1526, 1994PubMedGoogle Scholar
  29. 29.
    Collado A, de Andres A, Canadas E, Ruiz-Cabello F, Gomez O, Pedrinaci S, Garrido F: Characterization of CD44 antigen during lymphoid ontogeny. Immunobiology 183:1–11, 1991PubMedGoogle Scholar
  30. 30.
    Hale LP, Martin ME, McCollum DE, Nunley JA, Springer TA, Singer KH, Haynes BF: Immunohistologic analysis of the distribution of cell adhesion molecules within the inflammatory synovial microenvironment. Arth Rheum 32:22–30, 1989Google Scholar
  31. 31.
    McCachren SS: Expression of metalloproteinases and metalloproteinase inhibitor in human arthritic synovium. Arth Rheum 34: 1085–1093, 1991Google Scholar
  32. 32.
    Rooney M, Condell D, Quinlan W, Daly L, Whelan A, Feighery C, Bresnihan B: Analysis of the histologic variation of synovitis in rheumatoid arthritis. Arth Rheum 31:956–963, 1988Google Scholar
  33. 33.
    Kugelman LC, Ganguly S, Haggerty JG, Weissman SM, Milstone LM: The core protein of epican, a heparan sulfate proteoglycan on keratinocytes, is an alternative form of CD44. J Invest Dermatol 99:886–891, 1992PubMedGoogle Scholar
  34. 34.
    Haynes BF, Harden EA, Telen MJ, Hemler ME, Strominger JL, Palker TJ, Scearce RM, Eisenbarth GS: Differentiation of human T lymphocytes. I. Acquisition of a novel human cell surface protein (p80) during normal intrathymic T cell maturation. J Immunology 131:1195–1200, 1983Google Scholar
  35. 35.
    Telen MJ, Eisenbarth GS, Haynes BF: Human erythrocyte antigens: Regulation of expression of a novel red cell surface antigen by the inhibitor Lutheran In(Lu) gene. J Clin Invest 71:1878–1886, 1983PubMedGoogle Scholar
  36. 36.
    Dimitriu-Bona A, Burmester GR, Waters SJ, Winchester RJ: Human mononuclear phagocyte differentiation antigens. J Immunol 130:145–152, 1983PubMedGoogle Scholar
  37. 37.
    Stamenkovic I, Amiot M, Pesando JM, Seed B: A lymphocyte molecule implicated in lymph node homing is a member of the cartilage link protein family. Cell 56:1057–1062, 1989PubMedGoogle Scholar
  38. 38.
    Goldstein LA, Zhou DF, Picker LJ, Minty CN, Bargatze RF, Ding JF, Butcher EC: A human lymphocyte homing receptor, the hermes antigen, is related to cartilage proteoglycan core and link proteins. Cell 56:1063–1072, 1989PubMedGoogle Scholar
  39. 39.
    Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159, 1987PubMedGoogle Scholar
  40. 40.
    Wikstrand CJ, Hale LP, Batra SK, Hill MJ, Humphrey PA, Kurpad SN, McLendon RE, Moscatello D, Pegram CN, Reist CJ, Traweek ST, Wong AJ, Zalutsky MR, Bigner DD: Monoclonal antibodies against EFGRvIII are tumor specific and react with breast and lung carcinomas and malignant gliomas. Cancer Res 55:3140–3148, 1995PubMedGoogle Scholar
  41. 41.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Plainview, NY, Cold Spring Harbor Laboratory Press, 1989Google Scholar
  42. 42.
    Stamenkovic I, Aruffo A, Amiot M, Seed B: The hematopoietic and epithelial forms of CD44 are distinct polypeptides with different adhesion potentials for hyaluronate-bearing cells. EMBO J 10:343–348, 1991PubMedGoogle Scholar
  43. 43.
    Koopman WJ, Gay S: Do nonimmunologically mediated pathways play a role in the pathogenesis of rheumatoid arthritis? Rheum Clin North Am 19:107–122, 1993Google Scholar
  44. 44.
    Kalomiris EL, Bourguignon LYW: Lymphoma protein kinase C is associated with the transmembrane glycoprotein, gp85, and may function in gp85 ankyrin binding. J Biol Chem 264:8113–8119, 1989PubMedGoogle Scholar
  45. 45.
    Bourguignon LY, Lokeshwar VB, He J, Chen X, Bourguignon GJ: A CD44-like endothelial cell transmembrane glycoprotein (GP116) interacts with extracellular matrix and ankyrin. Mol Cell Biol 12:4464–4471, 1992PubMedGoogle Scholar
  46. 46.
    Lokeshwar VB, Bourguignon LY: The lymphoma transmembrane glycoprotein GP85 (CD44) is a novel guanine nucleotide-binding protein which regulates GP85 (CD44)-ankyrin interaction. J Biol Chem 267:22073–22078, 1992PubMedGoogle Scholar
  47. 47.
    Aruffo A, Stamenkovic I, Melnick M, Underhill CB, Seed B: CD44 is the principal cell surface receptor for hyaluronate. Cell 61:1303–1313, 1990PubMedGoogle Scholar
  48. 48.
    Miyake K, Underhill CB, Lesley J, Kincade PW: Hyaluronate can function as a cell adhesion molecule and CD44 participates in hyaluronate recognition. J Exp Med 172:69–75, 1990PubMedGoogle Scholar
  49. 49.
    Hiro D, Ho A, Matsuta K, Mori Y: Hyaluronic acid is an endogenous inducer of interleukin-1 production by human monocytes and rabbit macrophages. Biochem Biophys Res Commun 140:715–722, 1986PubMedGoogle Scholar
  50. 50.
    Noble PW, Lake FR, Henson PM, Riches DW: Hyaluronate activation of CD44 induces insulin-like growth factor-1 expression by a tumor necrosis factor-alpha-dependent mechanism in murine macrophages. J Clin Invest 91:2368–2377, 1993PubMedGoogle Scholar
  51. 51.
    Schubert M, Hammerman D: The functioning of the diffuse macromolecules of joints. Bull Rheum Dis 14:345–348, 1964PubMedGoogle Scholar
  52. 52.
    Castor CW, Prince RK, Hazelton MJ: Hyaluronic acid in human synovial effusions: a sensitive indicator of altered connective tissue cell function during inflammation. Arth Rheum 9:783–794, 1966Google Scholar
  53. 53.
    Culty M, Nguyen HA, Underhill CB: The hyaluronan receptor (CD44) participates in the uptake and degradation of hyaluronan. J Cell Biol 116:1055–1062, 1992PubMedGoogle Scholar
  54. 54.
    Camp RL, Kraus TA, Pure E: Variations in the cytoskeletal interaction and posttranslational modification of the CD44 homing receptor in macrophages. J Cell Biol 115:1283–1292, 1991PubMedGoogle Scholar
  55. 55.
    Sy MS, Guo YJ, Stamenkovic I: Distinct effects of two CD44 isoforms on tumor growth in vivo. J Exp Med 174:859–866, 1991PubMedGoogle Scholar
  56. 56.
    He Q, Lesley J, Hyman R, Ishihara K, Kincade PW: Molecular isoforms of murine CD44 and evidence that the membrane proximal domain is not critical for hyaluronate recognition. J Cell Biol 119:1711–1719, 1992PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1995

Authors and Affiliations

  • Laura P. Hale
    • 1
  • Barton F. Haynes
    • 2
  • S. Spence McCachren
    • 3
  1. 1.Department of PathologyDuke University Medical CenterDurham
  2. 2.Department of Medicine, Division of Rheumatology, Allergy and Clinical immunologyDuke University Arthritis CenterDurham
  3. 3.Department of Medicine, Division of Hematology and Medical OncologyDuke University Arthritis Center, and Geriatric Research, Education and Clinical Center, Veterans Affairs Medical CenterDurham

Personalised recommendations