Spatial distribution of L-selectin (CD62L) on human lymphocytes and transfected murine L1-2 cells
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We have examined the topographical distribution of L-selectin on surface membrane domains of human lymphocytes and murine L1-2 cells transfected to express human L-selectin. L-selectin was immunolocalized using murine monoclonal DREG 200 Fab antibody and a 12 nm colloidal gold-conjugated secondary antibody. Cell surface morphology and surface distribution of gold-labelled L-selectin were visualized using backscatter electron images obtained by high-resolution, field emission scanning electron microscopy. The topographical morphologies of lymphocytes of both types were complex. The surface of human lymphocytes was composed of both microvilli and ruffles; that of the murine cells was composed of long microvilli and few, if any, ruffles. L-selectin on human lymphocytes was observed primarily as focal clusters on the apical surfaces of ruffles and microvilli. Similarly, on the transfected murine cells, L-selectin was detected predominantly on the apical surface of microvilli. We conclude that L-selectin has a common spatial distribution and clustered organization on all leukocytes examined to-date, and that these features of receptor expression likely facilitate rolling of circulating leukocytes on the endothelial surface.
KeywordsField Emission Scanning Electron Microscopy Surface Membrane Field Emission Scanning Emission Scanning Electron Backscatter Electron
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- Berlin, C., Bargatze, R. F., Campbell, J. J., Von Andrian, U. H., Szabo M. C., Hasslen, S. R., Nelson, R. D., Berg, E. L., Erlandsen, S. L. &Butcher, E. C. Integrin-mediated lymphocyte attachment and rolling under physiologic flow.Cell (in press).Google Scholar
- Cohnheim,J. (1889)Lectures on General Pathology: a Handbook for Practitioners and Students. London: The New Syndenham Society.Google Scholar
- Kishimoto,T. K. (1991) A dynamic model for neutrophil localization to inflammatory sites.J. NIH Res. 3, 75–7.Google Scholar
- Laudanna,C.,Constantin,G.,Baron,P.,Scarpini,E.,Scarloato,G.,Cabrini,G.,Dechecchi,C.,Rossi,F.,Cassetella,M. A. &Berton,D. (1994) Sulfatides trigger increase of cytosolic free calcium and enhanced expression of tumor necrosis factor-α and interleukin-8 mRNA in human neutrophils.J. Biol. Chem. 269, 4021–6.PubMedGoogle Scholar
- Ley,K.,Tedder,T. F. &Kansas,G. S. (1991) L-selectin can mediate leukocyte rolling in untreated mesenteric venules in vivo independent of E or P selectin.Blood 82, 1632–8.Google Scholar
- VonAndrian,U. H.,Chambers,J. D.,Mcevoy,L. M.,Bargatze,R. F.,Arfors,K. E. &Butcher,E. C. (1991) Two-step model of leukocyte-endothelial cell interaction in inflammation: distinct roles for LECAM-1 and the leukocyte β2 integrins in vivo.Proc. Natl Acad. Sci. USA 88, 7538–42.Google Scholar
- VonAndrian,U. H.,Chambers,J. D.,Berg,E. L.,Michie,S. A.,Brown,D. A.,Karolak,D.,Ramezani,L.,Berger,E. M.,Arfors,K. E. &Butcher,E. C. (1993) L-selectin mediates neutrophil rolling in inflamed venules through sialyl Lewisx-dependent and independent recognition pathways.Blood 82, 182–91.Google Scholar
- Von Andrian, U. H., Hasslen, S. R., Nelson, R. D., Erlandsen, S. L. &Butcher, E. C. Mechanisms and consequences of microvillous receptor presentation in leukocyte adhesion under flow.Cell (submitted).Google Scholar