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. 1998 Aug;94(4):523–528. doi: 10.1046/j.1365-2567.1998.00551.x

Characterization of E-selectin-binding epitopes expressed by skin-homing T cells.

R Priest 1, M I Bird 1, R Malhotra 1
PMCID: PMC1364230  PMID: 9767440

Abstract

The glycoprotein counter-receptors for E-selectin borne on skin-homing T cells are poorly defined. In this study we have used flow cytometry to investigate the surface expression of potential carbohydrate ligands for E-selectin on HUT78, a skin-homing cutaneous T-cell lymphoma. These cells possessed high surface expression of the KM-93 epitope but not HECA 452 or CSLEX1 epitopes. The KM-93 antibody also blocked the binding of HUT78 cells to E-selectin. All these antibodies are reported to recognize sialyl Lewis X (sLex)-like molecules. Using an E-selectin affinity matrix, the main glycoprotein isolated from HUT78 cells was a molecular species of 90 000 MW. Other minor species of molecular weights 40 000, 60 000, 100 000, 120 000 and 200 000 were also identified as potential counter-receptors for E-selectin. Four of the purified counter-receptors (90 000, 100 000, 120 000 and 200 000 MW) stained positive with the KM-93 antibody. Immunoblot analysis of these purified glycoproteins established the identity of the 90 000 MW glycoprotein as l-selectin. Furthermore, an anti-l-selectin antibody inhibited the binding of HUT78 cells to E-selectin, probably by steric inhibition of the carbohydrate ligand for E-selectin that is borne on the C-type lectin domain of l-selectin. These results suggest that a carbohydrate epitope on l-selectin may act as a ligand for E-selectin on skin-homing T cells.

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Selected References

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  1. Anostario M., Jr, Huang K. S. Development of scintillation proximity assays for E-selectin and their application in testing potential antagonists. Anal Biochem. 1995 Nov 20;232(1):122–128. doi: 10.1006/abio.1995.9951. [DOI] [PubMed] [Google Scholar]
  2. Berg E. L., Yoshino T., Rott L. S., Robinson M. K., Warnock R. A., Kishimoto T. K., Picker L. J., Butcher E. C. The cutaneous lymphocyte antigen is a skin lymphocyte homing receptor for the vascular lectin endothelial cell-leukocyte adhesion molecule 1. J Exp Med. 1991 Dec 1;174(6):1461–1466. doi: 10.1084/jem.174.6.1461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bevilacqua M. P., Pober J. S., Mendrick D. L., Cotran R. S., Gimbrone M. A., Jr Identification of an inducible endothelial-leukocyte adhesion molecule. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9238–9242. doi: 10.1073/pnas.84.24.9238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dohi T., Nemoto T., Ohta S., Shitara K., Hanai N., Nudelman E., Hakomori S., Oshima M. Different binding properties of three monoclonal antibodies to sialyl Le(x) glycolipids in a gastric cancer cell line and normal stomach tissue. Anticancer Res. 1993 Sep-Oct;13(5A):1277–1282. [PubMed] [Google Scholar]
  5. Game S. M., Rajapurohit P. K., Clifford M., Bird M. I., Priest R., Bovin N. V., Nifant'ev N. E., O'Beirne G., Cook N. D. Scintillation proximity assay for E-, P-, and L-selectin utilizing polyacrylamide-based neoglycoconjugates as ligands. Anal Biochem. 1998 Apr 10;258(1):127–135. doi: 10.1006/abio.1998.2576. [DOI] [PubMed] [Google Scholar]
  6. Groves R. W., Allen M. H., Barker J. N., Haskard D. O., MacDonald D. M. Endothelial leucocyte adhesion molecule-1 (ELAM-1) expression in cutaneous inflammation. Br J Dermatol. 1991 Feb;124(2):117–123. doi: 10.1111/j.1365-2133.1991.tb00419.x. [DOI] [PubMed] [Google Scholar]
  7. Jones W. M., Watts G. M., Robinson M. K., Vestweber D., Jutila M. A. Comparison of E-selectin-binding glycoprotein ligands on human lymphocytes, neutrophils, and bovine gamma delta T cells. J Immunol. 1997 Oct 1;159(7):3574–3583. [PubMed] [Google Scholar]
  8. Kriegel T. M., Rush J., Vojtek A. B., Clifton D., Fraenkel D. G. In vivo phosphorylation site of hexokinase 2 in Saccharomyces cerevisiae. Biochemistry. 1994 Jan 11;33(1):148–152. doi: 10.1021/bi00167a019. [DOI] [PubMed] [Google Scholar]
  9. Kuzel T. M., Roenigk H. H., Jr, Rosen S. T. Mycosis fungoides and the Sézary syndrome: a review of pathogenesis, diagnosis, and therapy. J Clin Oncol. 1991 Jul;9(7):1298–1313. doi: 10.1200/JCO.1991.9.7.1298. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Levinovitz A., Mühlhoff J., Isenmann S., Vestweber D. Identification of a glycoprotein ligand for E-selectin on mouse myeloid cells. J Cell Biol. 1993 Apr;121(2):449–459. doi: 10.1083/jcb.121.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Life P., Gauchat J. F., Schnuriger V., Estoppey S., Mazzei G., Durandy A., Fischer A., Bonnefoy J. Y. T cell clones from an X-linked hyper-immunoglobulin (IgM) patient induce IgE synthesis in vitro despite expression of nonfunctional CD40 ligand. J Exp Med. 1994 Nov 1;180(5):1775–1784. doi: 10.1084/jem.180.5.1775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Malhotra R., Taylor N. R., Bird M. I. Anionic phospholipids bind to L-selectin (but not E-selectin) at a site distinct from the carbohydrate-binding site. Biochem J. 1996 Feb 15;314(Pt 1):297–303. doi: 10.1042/bj3140297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Misugi E., Kawamura N., Imanishi N., Tojo S. J., Morooka S. Sialyl Lewis X moiety on rat polymorphonuclear leukocytes responsible for binding to rat E-selectin. Biochem Biophys Res Commun. 1995 Oct 13;215(2):547–554. doi: 10.1006/bbrc.1995.2499. [DOI] [PubMed] [Google Scholar]
  15. Munro J. M., Lo S. K., Corless C., Robertson M. J., Lee N. C., Barnhill R. L., Weinberg D. S., Bevilacqua M. P. Expression of sialyl-Lewis X, an E-selectin ligand, in inflammation, immune processes, and lymphoid tissues. Am J Pathol. 1992 Dec;141(6):1397–1408. [PMC free article] [PubMed] [Google Scholar]
  16. Noorduyn L. A., Beljaards R. C., Pals S. T., van Heerde P., Radaszkiewicz T., Willemze R., Meijer C. J. Differential expression of the HECA-452 antigen (cutaneous lymphocyte associated antigen, CLA) in cutaneous and non-cutaneous T-cell lymphomas. Histopathology. 1992 Jul;21(1):59–64. doi: 10.1111/j.1365-2559.1992.tb00343.x. [DOI] [PubMed] [Google Scholar]
  17. Ohta S., Hanai N., Habu S., Nishimura T. Expression of sialyl Lewis(x) antigen on human T cells. Cell Immunol. 1993 Oct 15;151(2):491–497. doi: 10.1006/cimm.1993.1258. [DOI] [PubMed] [Google Scholar]
  18. Phillips M. L., Nudelman E., Gaeta F. C., Perez M., Singhal A. K., Hakomori S., Paulson J. C. ELAM-1 mediates cell adhesion by recognition of a carbohydrate ligand, sialyl-Lex. Science. 1990 Nov 23;250(4984):1130–1132. doi: 10.1126/science.1701274. [DOI] [PubMed] [Google Scholar]
  19. Picker L. J., Kishimoto T. K., Smith C. W., Warnock R. A., Butcher E. C. ELAM-1 is an adhesion molecule for skin-homing T cells. Nature. 1991 Feb 28;349(6312):796–799. doi: 10.1038/349796a0. [DOI] [PubMed] [Google Scholar]
  20. Picker L. J., Warnock R. A., Burns A. R., Doerschuk C. M., Berg E. L., Butcher E. C. The neutrophil selectin LECAM-1 presents carbohydrate ligands to the vascular selectins ELAM-1 and GMP-140. Cell. 1991 Sep 6;66(5):921–933. doi: 10.1016/0092-8674(91)90438-5. [DOI] [PubMed] [Google Scholar]
  21. Pinola M., Renkonen R., Majuri M. L., Tiisala S., Saksela E. Characterization of the E-selectin ligand on NK cells. J Immunol. 1994 Apr 1;152(7):3586–3594. [PubMed] [Google Scholar]
  22. Pitzalis C., Cauli A., Pipitone N., Smith C., Barker J., Marchesoni A., Yanni G., Panayi G. S. Cutaneous lymphocyte antigen-positive T lymphocytes preferentially migrate to the skin but not to the joint in psoriatic arthritis. Arthritis Rheum. 1996 Jan;39(1):137–145. doi: 10.1002/art.1780390118. [DOI] [PubMed] [Google Scholar]
  23. Priest R., Nawaz S., Green P. M., Bird M. I. Adhesion of eosinophils to E- & P-selectin. Biochem Soc Trans. 1995 May;23(2):162S–162S. doi: 10.1042/bst023162s. [DOI] [PubMed] [Google Scholar]
  24. Rossiter H., van Reijsen F., Mudde G. C., Kalthoff F., Bruijnzeel-Koomen C. A., Picker L. J., Kupper T. S. Skin disease-related T cells bind to endothelial selectins: expression of cutaneous lymphocyte antigen (CLA) predicts E-selectin but not P-selectin binding. Eur J Immunol. 1994 Jan;24(1):205–210. doi: 10.1002/eji.1830240132. [DOI] [PubMed] [Google Scholar]
  25. Santamaria Babi L. F., Picker L. J., Perez Soler M. T., Drzimalla K., Flohr P., Blaser K., Hauser C. Circulating allergen-reactive T cells from patients with atopic dermatitis and allergic contact dermatitis express the skin-selective homing receptor, the cutaneous lymphocyte-associated antigen. J Exp Med. 1995 May 1;181(5):1935–1940. doi: 10.1084/jem.181.5.1935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Steeber D. A., Green N. E., Sato S., Tedder T. F. Lyphocyte migration in L-selectin-deficient mice. Altered subset migration and aging of the immune system. J Immunol. 1996 Aug 1;157(3):1096–1106. [PubMed] [Google Scholar]
  27. Tang M. L., Hale L. P., Steeber D. A., Tedder T. F. L-selectin is involved in lymphocyte migration to sites of inflammation in the skin: delayed rejection of allografts in L-selectin-deficient mice. J Immunol. 1997 Jun 1;158(11):5191–5199. [PubMed] [Google Scholar]
  28. Tiemeyer M., Swiedler S. J., Ishihara M., Moreland M., Schweingruber H., Hirtzer P., Brandley B. K. Carbohydrate ligands for endothelial-leukocyte adhesion molecule 1. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1138–1142. doi: 10.1073/pnas.88.4.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zöllner O., Lenter M. C., Blanks J. E., Borges E., Steegmaier M., Zerwes H. G., Vestweber D. L-selectin from human, but not from mouse neutrophils binds directly to E-selectin. J Cell Biol. 1997 Feb 10;136(3):707–716. doi: 10.1083/jcb.136.3.707. [DOI] [PMC free article] [PubMed] [Google Scholar]

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