%0 Journal Article %J Eur J Immunol. %D 1995 %T Heterogenous glycosylation of ICAM-3 and lack of interaction with Mac-1 and p150,95. %A de Fougerolles, A.R. %A Diamond,M.S. %A Springer, T.A. %X

Intercellular adhesion molecule (ICAM)-1, ICAM-2, and ICAM-3 have been identified as counter-receptors for the leukocyte integrin lymphocyte function-associated antigen 1 (LFA-1). The other leukocyte integrins, Mac-1 and p150,95, also interact with ICAM-1. ICAM-1 and ICAM-3 are highly homologous, and an undefined ligand for Mac-1 is present on neutrophils where ICAM-3 is well expressed. In addition, glycosylation has been shown to affect the interaction of ICAM-1 with Mac-1. We therefore sought to characterize ICAM-3 heterogeneity and determine whether ICAM-3 was a ligand for either Mac-1 or p150,95. Despite extensive differences in N-linked glycosylation, ICAM-3 purified from lymphoid cells and from neutrophils supports adhesion of LFA-1-bearing cells equally well; however, neither supports adhesion of Mac-1 or p150,95-expressing chinese hamster ovary cell transfectants. Similarly, purified Mac-1 does not support adhesion of ICAM-2 or ICAM-3-expressing L cell transfectants. ICAM-3 on neutrophils does not participate in Mac-1-dependent homotypic aggregation. Thus, ICAM-3 is not a counter-receptor for either Mac-1 or p150,95.

%B Eur J Immunol. %V 25 %P 1008-12 %G eng %N 4 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Stimulation of binding of LFA-1 bearing cells to ICAM-1 and ICAM-3 %A Petruzzelli,L. %A Maduzia,L. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1594-1595 %G eng %1 305 %F 305 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Localization of the LFA-1 binding site on ICAM-3 (CD50) by epitope mapping and deletion mutagenesis %A Klickstein,L.B. %A de Fougerolles,A.R. %A York,M.R. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1563-1566 %G eng %1 299 %F 299 %0 Journal Article %J J. Immunol. %D 1995 %T Activation of LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18) mimicked by an antibody directed against CD18 %A Petruzzelli,L. %A Maduzia,L. %A Springer, T.A. %X

The beta 2-integrin (CD18) family members bind to their ligands subsequent to activation of a number of well defined and diverse signal transduction pathways. The precise molecular changes associated with activation of the integrin family members have remained elusive. Here, we characterize a monoclonal, CBR LFA-1/2, that binds to the beta 2-subunit and is able to mimic activation induced upon stimulation by phorbol esters. The Ab induces binding of the LFA-1-expressing cell line, JY, to ICAM-1 (CD54) and ICAM-3 (CD50). Activation of binding by this Ab is independent of Fc interactions and does not occur through cross-linking at the cell surface, because the Fab fragment of the Ab is able to modulate the same effect. Stimulation of neutrophils with CBR LFA-1/2 induces binding to ICAM-1 through activation of both LFA-1 and Mac-1. Activation of Mac-1 by CBR LFA-1/2 was further confirmed by stimulation of neutrophil binding to fibrinogen, a ligand for Mac-1. CBR LFA-1/2 lowers by 10-fold the concentration of Mg2+ required to achieve maximal binding of LFA-1 to ICAM-1. It therefore appears that CBR LFA-1/2 induces a conformational change that directly increases the avidity of beta 2-integrins for ligands.

%B J. Immunol. %V 155 %P 854-866 %G eng %N 2 %1 278 %! J. Immunol. %F 278 %0 Journal Article %J Int. Immunol. %D 1995 %T Activation of natural killer cells by the mAb YTA-1 that recognizes leukocyte function-associated antigen-1 %A Sugie, K. %A Nakamura, K. %A Teshigawara, K. %A Diamond,M.S. %A Springer, T.A. %A Nakamura, Y. %A Leonard, W.J. %A Uchida, A. %A Yodoi, J. %K ACTIVATION %X

The mAb YTA-1, which brightly stains CD3-CD16+ large granular lymphocytes (LGL)/natural killer (NK) cells and CD8+ T cells by immunofluorescence, is specific for leukocyte function-associated antigen (LFA)-1. Some mAbs recognizing the LFA-1 alpha chain (CD11a) or LFA-1 beta chain (CD18) inhibited the binding of YTA-1 to peripheral blood mononuclear cells. YTA-1 mAb could be chemically cross-linked to 170 and 96 kDa molecules, whose molecular weights correspond to those of LFA-1 alpha and beta respectively. YTA-1 bound to COS-7 cells co-transfected with CD11a and CD18 cDNAs, but not to untransfected cells. Reactivities of YTA-1 to K562 cells transfected with LFA-1 alpha and beta (CD11a/CD18) cDNAs and to CHO cells transfected with Mac-1 (CD11b/CD18) or p150, 95 (CD11c/CD18) cDNAs strongly suggest that YTA-1 recognizes either LFA-1 alpha or an epitope formed by a combination of LFA-1 alpha and beta. Treatment of fresh CD3-CD16+ LGL with YTA-1 augmented cytolytic activity and induced proliferation. F(ab')2 fragments of YTA-1 augmented NK cytotoxicity, indicating that the NK activating signal was transmitted through LFA-1 without involvement of Fc gamma receptor III. In contrast, the other mAbs against LFA-1 could not activate NK cells. These results collectively indicate that YTA-1 recognizes a unique epitope of LFA-1, which is involved in activation of fresh NK cells.

%B Int. Immunol. %V 7 %P 763-769 %G eng %N 5 %1 276 %! Int. Immunol. %F 9222 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Adhesion structure subpanel 1, E rosetting/GPI anchor: CD2, CD48, CD55, CD58, CD99 and CDw108 %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,C. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1468-1471 %G eng %1 283 %F 283 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Adhesion structure subpanel 2, selectins: CD62E, CD62L, and CD62P %A Diacovo,T. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1498-1499 %G eng %1 290 %F 290 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Adhesion structure subpanel 4: CD50 (ICAM-3), CD54 (ICAM-1), and CD102 (ICAM-2) %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1542-1545 %G eng %1 294 %F 294 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Adhesion structure subpanel 5, leukocyte integrins: CD11a, CD11b, CD11c, CD18 %A Petruzzelli,L. %A Luk,J. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1581-1585 %G eng %1 300 %F 300 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Adhesion structure subpanels 7 and 8, β3, β4, β7 integrins and novel functional antigens: CD51, CD61, CD103, and CD104 %A Wong,D.A. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1655-1659 %G eng %1 308 %F 308 %0 Book Section %B Leukocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Adhesion Structures: Section Report %A Springer, T.A. %A Luther,E. %A Klickstein,L.B. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leukocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1443-1467 %G eng %1 282 %F 282 %0 Journal Article %J J. Biol. Chem. %D 1995 %T A binding interface on the I domain of lymphocyte function associated antigen-1 (LFA-1) required for specific interaction with intercellular adhesion molecule 1 (ICAM-1) %A Huang, C %A Springer, T.A. %K ICAM1 %X

Previous studies have shown that lymphocyte function-associated antigen-1 (LFA-1) molecules containing the human alpha (CD11a) and human beta (CD18) subunits but not the murine alpha and human beta subunits can bind to human intercellular adhesion molecule 1 (ICAM-1). Using human/mouse LFA-1 alpha subunit chimeras, we mapped regions required for binding to ICAM-1 N-terminal to amino acid (aa) residue 350. Ligand binding sites were mapped in greater detail by scanning this region with murine sequences from 56 down to 17 aa in length and finally by introducing single or few murine aa residue replacements into the human sequence. Replacement of two non-contiguous regions of aa residues 119-153 and 218-248 in the me domain with the corresponding mouse sequences abolished most binding to human ICAM-1, without affecting alpha beta subunit association or expression on the surface of transfected COS cells. Specific residues within the I domain found to be important were Met-140, Glu-146, Thr-243, and Ser-245. Using the recently solved structure of the Mac-1 (CD11b) I domain as a model (Lee, J.-O., Rieu, P., Arnaout, M.A., and Liddington, R. (1995) Cell 80, 631-638), these residues are shown to be located on the surface of the I domain surrounding the site to which Mg2+ is chelated, and fine a ligand binding interface. Mapping of the epitopes of a panel of mouse anti-human and rat anti-mouse monoclonal antibodies gave concordant results. Epitopes were mapped to two different regions in the N-terminal domain, four regions within the I domain, and two regions between the I domain and the EF hand-like repeats. Monoclonal antibodies to epitopes within the mid- to C-terminal portion of the I domain and the N-terminal portion of the region between the I domain and the EF hand-like repeats gave good inhibition of LFA-1-dependent homotypic aggregation with cells that express either ICAM-1 or ICAM-3 as the major LFA-1 ligand.

%B J. Biol. Chem. %V 270 %P 19008-19016 %G eng %N 32 %1 316 %! J. Biol. Chem. %F 316 %0 Journal Article %J Eur. J. Immunol. %D 1995 %T C-C chemokines, but not the C-X-C chemokines Il-8 and IP-10, stimulate transendothelial chemotaxis of T lymphocytes %A Roth,S.J. %A Carr,M.W. %A Springer, T.A. %X

Eight chemokines were tested for ability to elicit transendothelial chemotaxis of unstimulated peripheral blood T lymphocytes. The C-C chemokines monocyte chemotactic protein (MCP)-2, MCP-3, RANTES, macrophage inflammatory protein (MIP)-1 alpha, MIP-1 beta, and, as previously described, MCP-1 induced significant, dose-dependent transendothelial chemotaxis of CD3+ T lymphocytes. In contrast, the C-X-C chemokines interleukin-8 (IL-8) and interferon-gamma inducible protein-10 (IP-10) failed to induce transendothelial chemotaxis of CD3+ T lymphocytes or T lymphocyte subsets. RANTES, MIP-1 alpha, and MIP-1 beta induced significant transendothelial chemotaxis of CD4+, CD8+, and CD45R0+ T lymphocyte subsets. Phenotyping of mononuclear cells that underwent transendothelial migration to MCP-2, MCP-3, RANTES, or MIP-1 alpha showed both monocytes and activated (CD26 high), memory-type (CD45R0+) T cells. Both CD4+ and CD8+ T lymphocytes were recruited, but not natural killer cells or significant numbers of B cells. MCP-2 was the only C-C chemokine tested that attracted a significant number of naive-type (CD45RA+) T lymphocytes. In the absence of endothelium, IL-8 but not IP-10 promoted modest but significant chemotoxis of CD3+ T lymphocytes. Our data support the hypothesis that C-C, not the C-X-C chemokines IL-8 or IP-10, promote transendothelial chemotaxis of T lymphocytes.

%B Eur. J. Immunol. %V 25 %P 3482-3488 %G eng %N 12 %1 326 %! Eur. J. Immunol. %F 326 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD102 (ICAM-2) cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1550-1552 %G eng %1 297 %F 297 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD103 (αE) cluster report %A Cepek,K.L. %A Wong,D.A. %A Brenner,M.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %K CD103 %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1666-1667 %G eng %1 311 %F 311 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD104 (β4) cluster report %A Wong,D.A. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L.Gilks,W. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1667-1668 %G eng %1 312 %F 312 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD11a cluster report %A Petruzzelli,L. %A Huang, C %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,C. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1586-1587 %G eng %1 301 %F 301 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD11b cluster report %A Luk,J. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1588-1590 %G eng %1 302 %F 302 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD11c cluster report %A Luk,J. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1590-1592 %G eng %1 303 %F 303 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD18 cluster report %A Petruzzelli,L. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1592-1593 %G eng %1 304 %F 304 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD48 cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1472-1473 %G eng %1 284 %F 284 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD50 (ICAM-3) cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1546-1547 %G eng %1 295 %F 295 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD51 (αV) and CD51/CD61 complex cluster report %A Wong,D.A. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1663-1664 %G eng %1 309 %F 309 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD54 (ICAM-1) cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1548-1550 %G eng %1 296 %F 296 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD55 cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1473-1474 %G eng %1 285 %F 285 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD58 cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1475-1476 %G eng %1 286 %F 286 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD59 cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1476-1477 %G eng %1 287 %F 287 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD61 (β3) cluster report %A Wong,D.A. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1664-1665 %G eng %1 310 %F 310 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD62E (E-selectin) cluster report %A Diacovo,T. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1501-1503 %G eng %1 292 %F 292 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD62L (L-selectin) cluster report %A Diacovo,T. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1503-1504 %G eng %1 293 %F 293 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CD62P (P-selectin) cluster report %A Diacovo,T. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1500-1501 %G eng %1 291 %F 291 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T CDw108 cluster report %A Klickstein,L.B. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1477-1478 %G eng %1 288 %F 288 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Characterization of the ICAM-3/LFA-1 interaction and its role in immune responses %A de Fougerolles, A.R. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1557-1559 %G eng %1 298 %F 298 %0 Journal Article %J J. Immunol. %D 1995 %T Characterization of transendothelial chemotaxis of T lymphocytes %A Roth,S.J. %A Carr,M.W. %A Rose,S.S. %A Springer, T.A. %X

We have adapted a chemotaxis assay using human umbilical vein endothelial cell (HUVEC) monolayers on microporous membranes for studying lymphocyte transendothelial chemotaxis in vitro. Supernatants of peripheral blood mononuclear cells stimulated with phytohemagglutinin (PHA) were identified as an excellent source of lymphocyte chemoattractant activity. The activity in PHA supernatant typically caused 2-6% of peripheral blood lymphocytes (PBL) to transmigrate compared to 0.1-0.3% to media control. Checkerboard analysis demonstrated that transmigration was directional and not attributable to random locomotion. Purified T lymphocytes also underwent transendothelial chemotaxis to PHA supernatant. Using monoclonal antibodies to several human adhesion receptors, we found that the interaction between LFA-1 and ICAM-1/ICAM-2 was more important for transendothelial lymphocyte chemotaxis than the interaction between VLA-4 and VCAM-1. A monoclonal antibody to the beta 1 integrin subunit inhibited chemotaxis more than antibodies to the VLA alpha 2, alpha 3, alpha 4, or alpha 5 subunits. The transendothelial assay was used to guide purification of the lymphocyte chemoattractant activity, which we reported previously to be monocyte chemoattractant protein-1 (MCP-1) (Carr et al., Proc. Natl. Acad. Sci. USA (1994) 91, 3652). The adhesion molecules required for chemotaxis to MCP-1 were similar to those with PHA supernatant. The use of HUVEC in the assay enhances the signal-to-background ratio of chemotaxis and provides a model that is physiologically relevant to lymphocyte emigration from the bloodstream into sites of inflammation.

%B J. Immunol. %V 188 %P 97-116 %G eng %N 1 %1 325 %! J. Immunol. Methods %F 325 %0 Journal Article %J Immunity %D 1995 %T Cloning from purified high endothelial venule cells of hevin, a close relative of the antiadhesive extracellular matrix protein SPARC %A Girard, J-P. %A Springer, T.A. %X

High endothelial venules (HEV) in lymphoid tissues support high levels of lymphocyte extravasion from the blood. We purified high endothelial cells from human tonsils by immunomagnetic selection with MECA-79 MAb to construct an HEV cDNA library. Differential screening of this library using cDNA probes from HEV (plus) or flat-walled vessel (minus) endothelial cells allowed us to characterize a novel human cDNA expressed to high levels in HEV. The cDNA encodes a secreted acidic calcium-binding glycoprotein of 664 aa residues, designated hevin, exhibiting 62% identity with the antiadhesive extracellular matrix protein SPARC, over a region of 232 aa spanning more than four fifths of the SPARC coding sequence. The primary structure and sequence of hevin and similar to SPARC-like proteins from rat and quail, called SC1 or QR1. Hevin could contribute to the induction or maintenance of features of the HEV endothelium that facilitate lymphocyte migration.

%B Immunity %V 2 %P 113-122 %G eng %N 1 %1 267 %! Immunity %F 267 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Domain localization and correlation with inhibition of function of workshop CD11a mAb %A Huang, C %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1595-1597 %G eng %1 306 %F 306 %0 Journal Article %J J. Leukoc. Biol. %D 1995 %T Expression of glycophosphatidylinositol (GPI)-anchored and non-anchored isoforms of vascular cell adhesion molecule 1 in murine stromal and endothelial cells %A Kinashi,T. %A St.Pierre,Y. %A Springer, T.A. %K CD106 %K CD29 %K CD49d %X

Monoclonal antibodies to murine vascular cell adhesion molecule-1 (VCAM-1, CD106) revealed not only the expected VCAM-1 molecule with an apparent molecular weight of 100 kDa, but also a molecule with a smaller size of 46 kDa in stromal cells and stimulated endothelial cells. Peptide mapping suggested the 46 kDa and 100 kDa proteins were closely related. The 46 kDa, but not 100 kDa protein, was cleaved from the cell surface with phosphatidylinositol-specific phospholipase C (PI-PLC), showing that the 46 kDa protein was a GPI-linked molecule. The 46 kDa and 100 kDa isoforms of VCAM-1 were shown to be N-glycosylated, have similar kinetics of biosynthesis, and to be partially shed from the cell surface with a slight reduction of size. TNF-alpha induced both isoforms of VCAM-1 with a similar time course of appearance on the surface of endothelial cells. The relative amounts of the 46 kDa and 100 kDa isoforms depended on the cell type examined. The GPI-anchored isoform is functionally important, because on a cell on which it was expressed almost as well as the 100 kDa isoform, treatment with PI-PLC reduced VLA-4-dependent conjugate formation.

%B J. Leukoc. Biol. %V 57 %P 168-173 %G eng %N 1 %1 266 %! J. Leukoc. Biol. %F 9920 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Expression of sialomucin CD34 by high endothelial venules (HEV) in human tonsils %A Girard, J-P. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J.M. %E Kishimoto,T. %E Morimoto,C. %E Ritz,J. %E Shaw,S. %E Silverstein,R.L. %E Springer, T.A. %E Tedder,T.F. %E Todd,R.F. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1801-1803 %G eng %1 314 %F 314 %0 Journal Article %J J. Immunol. %D 1995 %T Glycolipid ligands for selectins support leukocyte tethering and rolling under physiologic flow conditions %A Alon, R. %A Feizi,T. %A Yuen,C-T. %A Fuhlbrigge,R.C. %A Springer, T.A. %X

Selectin interactions with glycolipids have been examined previously under static conditions, whereas physiologic interactions mediated by selectins take place under flow. We find that under physiologic flow conditions, sialyl Lewis(x) (sLe(x)) glycolipid and sialyl Lewisa (sLe(a)) neoglycolipid support tethering and rolling adhesions of Chinese hamster ovary (CHO) cells expressing E-selectin and lymphoid and myeloid cells expressing L-selectin. These selectin-mediated adhesions persist at the highest shear stresses that occur in postcapillary venules in vivo and occur at lower site densities than found for sLe(x) on neutrophils. The interactions are Ca(2+)-dependent and can be specifically and completely blocked with anti-selectin mAbs. Asialo nonfucosylated glycolipids are inactive, and sulfatide supports weak tethering, but not rolling, of L-selectin-expressing cells. Rolling velocities and resistance to detachment are related to the glycolipid site density and fall within the range measured for neutrophil and myeloid cell rolling on substrates containing purified selectins. These observations are the first indication that glycolipids can interact with selectins in physiologic flow conditions, and can contribute to rolling adhesions.

%B J. Immunol. %V 154 %P 5356-5366 %G eng %N 10 %1 275 %! J. Immunol. %F 275 %0 Journal Article %J J. Cell Biol. %D 1995 %T Heparin is an adhesive ligand for the leukocyte integrin Mac-1 (CD11b/CD18) %A Diamond,M.S. %A Alon, R. %A Parkos,C.A. %A Quinn,M.T. %A Springer, T.A. %X

Previous studies have demonstrated that the leukocyte integrin Mac-1 adheres to several cell surface and soluble ligands including intercellular adhesion molecule-1, fibrinogen, iC3b, and factor X. However, experiments with Mac-1-expressing transfectants, purified Mac-1, and mAbs to Mac-1 indicate the existence of additional ligands. In this paper, we demonstrate a direct interaction between Mac-1 and heparan sulfate glycans. Heparin affinity resins immunoprecipitate Mac-1, and neutrophils and transfectant cells that express Mac-1 bind to heparin and heparan sulfate, but not to other sulfated glycosaminoglycans. Inhibition studies with mAbs and chemically modified forms of heparin suggest the I domain as a recognition site on Mac-1 for heparin, and suggest that either N- or O-sulfation is sufficient for heparin to bind efficiently to Mac-1. Under conditions of continuous flow in which heparins and E-selectin are cosubstrates, neutrophils tether to E-selectin and form firm adhesions through a Mac-1-heparin interaction.

%B J. Cell Biol. %V 130 %P 1473-1482 %G eng %N 6 %1 318 %! J. Cell Biol. %F 318 %0 Journal Article %J Immunol. Today %D 1995 %T High endothelial venules (HEVs): Specialized endothelium for lymphocyte migration %A Girard, J-P. %A Springer, T.A. %X

High endothelial venules (HEVs) are specialized postcapillary venules found in lymphoid tissues that support high levels of lymphocyte extravasation from the blood. Here, Jean-Philippe Girard and Timothy Springer highlight the unique properties of HEV endothelium, discuss the molecular mechanisms controlling HEV specialization and review evidence suggesting that HEVs could play an important role in the pathogenesis of chronic inflammatory diseases.

%B Immunol. Today %V 16 %P 449-457 %G eng %N 9 %1 317 %! Immunol. Today %F 317 %0 Journal Article %J Int. Immunol. %D 1995 %T ICAM-1 is required for T cell proliferation but not for anergy or apoptosis induced by Staphylococcus aureus enterotoxin B in vivo %A Gonzalo,J.A. %A Martinez-A,C. %A Springer, T.A. %A Gutierrez-Ramos,J-C. %X

The response of T lymphocytes to superantigens requires expression of the appropriate TCR V beta gene products as well as the establishment of cellular interactions mediated by adhesion molecules. To study the role of intercellular adhesion molecule (ICAM)-1 in the response in vivo to superantigens, we have analyzed the effects induced by the bacterial superantigen Staphylococcus aureus enterotoxin B (SEB) in mice which have been made genetically deficient in ICAM-1. SEB treatment of wild-type mice causes proliferation, deletion and anergy of the SEB-reactive V beta 8+ T cell population. Here we show that cellular interactions mediated by ICAM-1 are not essential for the induction of anergy or for the deletion of CD4+ V beta 8+ or CD8+ V beta 8+ T cells, but are required for the proliferation of these peripheral T lymphocytes. This is the first demonstration in vivo that the absence of the co-stimulatory signals provided by the interaction of ICAM-1 with its specific ligands impairs the proliferation of SEB-reactive T cells. Interestingly, our study showed that SEB-induced proliferation of CD8+ V beta 8+ T cells from lymph nodes (not from spleen) is independent of the interactions mediated by ICAM-1.

%B Int. Immunol. %V 7 %P 1691-1698 %G eng %N 10 %1 322 %! Int. Immunol. %F 322 %0 Journal Article %J Chem. Biol. %D 1995 %T Ideas crystallized on immunoglobulin superfamily-integrin interactions %A de Fougerolles,A. %A Springer, T.A. %X

Interactions between immunoglobulin superfamily (IgSF) members and integrins are central to lymphocyte homing, leukocyte emigration into tissues at inflammatory sites, and in cell-cell interactions that lead to immune responses. Recent X-ray crystal structures reveal that the interaction of a divalent cation found in the integrin structure with an acidic residue from the IgSF partner may be important in binding.

%B Chem. Biol. %V 2 %P 639-43 %G eng %N 10 %! Chem. Biol. %F 3643 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Identification of novel GPI-anchored antigens by analysis of GPI-anchor-deficient cells with mAb in the blind panel %A Klickstein,L.B. %A York,M.R. %A Luther,E. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1478-1481 %G eng %1 289 %F 289 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Identification of workshop endothelial section mAb that recognize novel glycosylphosphatidylinositol-anchored antigens %A Klickstein,L.B. %A York,M.R. %A Luther,E. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1850-1852 %G eng %1 315 %F 315 %0 Journal Article %J J. Cell Biol. %D 1995 %T The integrin VLA-4 supports tethering and rolling in flow on VCAM-1 %A Alon, R. %A Kassner,P.D. %A Carr,M.W. %A Finger,E.B. %A Hemler,M.E. %A Springer, T.A. %K 451 %X

Selectins have previously been shown to tether a flowing leukocyte to a vessel wall and mediate rolling. Here, we report that an intergrin, VLA-4, can also support tethering and rolling. Blood T lymphocytes and alpha 4 integrin-transfected cells can tether in shear flow, and then roll, through binding of the intergrin VLA-4 to purified VCAM-1 on the wall of a flow chamber. VLA-4 transfectants showed similar tethering and rolling on TNF-stimulated endothelium. Tethering efficiency, rolling velocity, and resistance to detachment are related to VCAM-1 density. Tethering and rolling did not occur on ICAM-1, fibronectin, or fibronectin fragments, and tethering did not require integrin activation or the presence of an alpha 4 cytoplasmic domain. Arrest of rolling cells on VCAM-1 occurred spontaneously, and/or was triggered by integrin activating agents Mn2+, phorbol ester, and mAb TS2/16. These agents, and the alpha 4 cytoplasmic domain, promoted increased resistance to detachment. Together the results show that VLA-4 is a versatile integrin that can mediate tethering, rolling, and firm arrest on VCAM-1.

%B J. Cell Biol. %V 128 %P 1243-1253 %G eng %N 6 %1 271 %! J. Cell Biol. %F 271 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Integrin β7 pre-CD report %A Wong,D.A. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1669-1670 %G eng %1 313 %F 313 %0 Journal Article %J J. Biol. Chem. %D 1995 %T Kinetics and thermodynamics of virus binding to receptor: Studies with rhinovirus, intercellular adhesion molecule-1 (ICAM-1), and surface plasmon resonance %A Casasnovas,J.M. %A Springer, T.A. %K ICAM1 %X

We have studied the kinetics and thermodynamics of a virus interacting with its receptor using human rhinovirus serotype 3 (HRV3), soluble intercellular adhesion molecule-1 (ICAM-1, CD54) containing Ig superfamily domains 1-5 (sICAM-1), and surface plasmon resonance. There were two classes of binding sites for sICAM-1 on HRV3, each comprising about 50% of the total sites, with association rate constants of 2450 +/- 300 and 134 +/- 11 M-1 s-1. These rates are low, consistent with binding to a relatively inaccessible site in the rhinovirus canyon. By contrast, three monoclonal antibodies bound to sICAM-1 with a single rate constant of 17,000-48,000 M-1 s-1. The dissociation rate constant for HRV3 was 1.7 +/- 0.1 x 10(-3) s-1, giving calculated dissociation constants of 0.7 +/- 0.1 and 12.5 +/- 1.2 microM. Agreement was good with saturation binding in solution, which showed two sites of similar abundance with KD of 0.55 +/- 0.2 and 5.7 +/- 2.0 microM. A bivalent chimera of ICAM-1 with the IgA1 Fc region bound with KD = 50 and 410 nM, showing 17-fold enhanced affinity. Lowering pH from 8.0 to 6.0 reduced affinity by approximately 50-fold, primarily by reducing the on rate. Thermodynamic measurements showed that binding of ICAM-1 to HRV3 is endothermic, by contrast to binding to monoclonal antibody. The heat that is absorbed of 3.5 and 6.3 kcal/mol for the two classes of ICAM-1 binding sites may contribute to receptor-mediated disruption of virions, which has an activation energy of about 42 kcal/mol.

%B J. Biol. Chem. %V 270 %P 13216-13224 %G eng %N 22 %1 277 %! J. Biol. Chem. %F 277 %0 Journal Article %J Nature %D 1995 %T Lifetime of the P-selectin: carbohydrate bond and its response to tensile force in hydrodynamic flow %A Alon, R. %A Hammer, D. A. %A Springer, T.A. %X

Selectins tether to the blood vessel wall leukocytes that are flowing in the bloodstream and support subsequent labile rolling interactions as the leukocytes are subjected to hydrodynamic drag forces. To support this rolling, selectins have been proposed to have rapid bond association and dissociation rate constants, and special mechanical properties linking tensile forces and bond dissociation. We have visualized transient tethering and release of neutrophils in hydrodynamic flow on lipid bilayers containing densities of P-selectin below those required to support rolling. We report here that transient tethers had first-order kinetics and other characteristics suggesting a unimolecular interaction between P-selectin and its glycoprotein ligand (PSGL-1). The unstressed dissociation constant (off rate) was 1 s-1. Hydrodynamic shear stresses of up to 1.1 dyn cm-2, corresponding to a force on the bond of up to 110 pN, increased the off rate only modestly, to 3.5 s-1. The data was adequately matched by a proposed equation relating off rate to the exponential of tensile force on the bond and the bond interaction distance, and gave a bond interaction distance of 0.5 A. This distance is compatible with hydrogen and metal coordination bonds between P-selectin and PSGL-1. Fast on and off rates, together with the high tensile strength of the selectin bond, appear necessary to support rolling at physiological shear stresses.

%B Nature %V 374 %P 539 %G eng %N 6522 %1 273 %! Nature %F 273 %0 Journal Article %J Blood %D 1995 %T Receptor tyrosine kinase stimulates cell-matrix adhesion by phosphatidylinositol 3 kinase and phospholipase C-γ1 pathways %A Kinashi,T. %A Escobedo,J.A. %A Williams,L.T. %A Takatsu,K. %A Springer, T.A. %X

Receptor tyrosine kinases are known to be important in growth and differentiation. We have recently found that c-kit, the tyrosine kinase receptor for steel factor, also regulates cell-matrix adhesion. Because Steel factor helps regulate cell migration and localization, this may be an important biologic function. Integrin adhesiveness is regulated within minutes by c-kit. The signaling pathways for tyrosine kinase stimulation of integrin adhesiveness and their relation to pathways that regulate growth and differentiation over much longer time periods remain uncharacterized. We have studied the effector pathways by which receptor tyrosine kinases regulate cell-matrix adhesion using wild-type and mutant forms of the platelet-derived growth factor (PDGF) receptor, which is closely related to c-kit. The PDGF receptor expressed in mast cells is as potent as c-kit in stimulating adhesion to fibronectin. We show that induction of adhesion is regulated through two independent pathways of phosphatidylinositol 3 kinase (PI3K) and phospholipase C-gamma 1 (PLC gamma)-protein kinase C by elimination of autophosphorylation sites required for activation of PI3K and PLC gamma or in combination with downregulation of protein kinase C or wortmannin. By contrast, a receptor mutated in both the PI3K and PLC gamma association sites can still stimulate mast cell growth, indicating a crucial role of these effector molecules in regulating adhesion rather than cell growth.

%B Blood %V 86 %P 2086-2090 %G eng %N 6 %1 319 %! Blood %F 319 %0 Journal Article %J Leuk. Lymphoma %D 1995 %T Regulation of cell-matrix adhesion by receptor tyrosine kinases %A Kinashi,T. %A Springer, T.A. %X

Cell-cell and cell-matrix adhesive interactions mediated by integrins play crucial roles in leukocyte migration to inflamed tissues, and also in cell migration during embryogenesis. Much remains to be learned about the molecular mechanisms of regulation of adhesion mediated by integrins. Recently we found that steel factor and c-kit induce adhesion to fibronectin by VLA-5 in mast cells. Activation of adhesiveness is transient, and occurs at concentrations of steel factor 100-fold lower than required for growth stimulation. This suggests that regulation of adhesion is an important biological function of steel factor and c-kit. Other receptor tyrosine kinases such as the PDGF receptor can substitute for c-kit. Signaling through receptor tyrosine kinases may offer a general mechanism for the regulation of integrin avidity.

%B Leuk. Lymphoma %V 18 %P 203-208 %G eng %N 3-4 %1 269 %! Leuk. Lymphoma %F 269 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Report on the CD34 cluster workshop %A Greaves,M.F. %A Titley,I. %A Colman,S.M. %A Buhring,H.-J. %A Campos,L. %A Castoldi,G.L. %A Garrido,F. %A Gaudernack,G. %A Girard, J-P. %A Ingles-Esteve,J. %A Invernizzi,R. %A Knapp,W. %A Lansdorp,P.M. %A Lanza,F. %A Merle-Beral,H. %A Parravicini,C. %A Razak,K. %A Ruiz-Cabello,F. %A Springer, T.A. %A Van Der Schoot,C.E. %A Sutherland,D.R. %E Schlossman,S. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,C. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %K CD34 %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 840-846 %G eng %1 281 %F 281 %0 Journal Article %J Eur. J. Immunol. %D 1995 %T Rolling of lymphocytes and neutrophils on peripheral node addressin and subsequent arrest on ICAM-1 in shear flow %A Lawrence,M.B. %A Berg,E.L. %A Butcher,E.C. %A Springer, T.A. %X

We studied leukocyte interactions in shear flow with peripheral lymph node addressin (PNAd), a mixture of glycoproteins expressed on high endothelial venules (HEV) that is required for lymphocyte homing and has been shown to contain a ligand for L-selectin. T lymphocytes and neutrophils tether and roll on plastic-immobilized PNAd and E-selectin at 1.8 dyn/cm2 wall shear stress, but fail to interact with immobilized ICAM-1, a ligand for LFA-1 and Mac-1, at the same flow rate. Cells roll faster on PNAd than on P-selectin or E-selectin. L-selectin mAb inhibit T lymphocyte and neutrophil tethering to PNAd, but do not inhibit T lymphocyte tethering to purified E-selectin. If allowed to interact with ICAM-1 under static conditions, phorbol ester-treated T lymphocytes, but not resting T lymphocytes, are able to form stationary adhesions that withstand the detachment force generated by 36 dyn/cm2 wall shear stress. In contrast, a wall shear stress of 7.3 dyn/cm2 detaches 50% of resting T lymphocytes bound to PNAd. Incubating T lymphocytes on PNAd and ICAM-1 does not result in adhesion strengthening, suggesting that adhesion through PNAd by L-selectin does not stimulate lymphocyte LFA-1 avidity for ICAM-1. Chemoattractant stimulation of neutrophils or phorbol ester stimulation of lymphoblasts rolling on coimmobilized PNAd and ICAM-1 results in rapid arrest and firm sticking, extending the model of sequential selectin-mediated rolling and subsequent integrin-mediated firm arrest to lymphocytes and ligands expressed on HEV.

%B Eur. J. Immunol. %V 25 %P 1025-1031 %G eng %N 4 %1 270 %! Eur. J. Immunol. %F 270 %0 Journal Article %J J. Cell Biol. %D 1995 %T Sialomucin CD34 is the major L-selectin ligand in human tonsil high endothelial venules %A Puri,K.D. %A Finger,E.B. %A Gaudernack,G. %A Springer, T.A. %X

Peripheral node addressin (PNAd) is a complex mixture of glycoproteins with L-selectin ligand activity that functions in lymphocyte homing. We have investigated the contribution of the sialomucin CD34 relative to other components of PNAd in lymphocyte tethering and rolling in in vitro laminar flow assays. PNAd was isolated with MECA-79 mAb-Sepharose from tonsillar stroma, and the CD34 component (PNAd,CD34+) and CD34-negative component (PNAd,CD34-) separated on CD34 mAb-Sepharose. Lymphocytes on the PNAd,CD34- fraction tether less efficiently, roll faster and are less resistant to shear detachment than on PNAd. The PNAd,CD34+ fraction constitutes about half the total functional activity. These studies show that CD34 is a major functional component of PNAd. Ligand activity in both the PNAd,CD34+ and PNAd,CD34- fractions is expressed on mucin-like domains, as shown with O-sialoglycoprotease. The CD34 component of PNAd has about four times higher tethering efficiency than total tonsillar CD34. CD34 from spleen shows no lymphocyte tethering. Although less efficient than the PNAd,CD34+ fraction from tonsil, CD34 from the KG1a hematopoietic cell line is functionally active as an L-selectin ligand despite lack of reactivity with MECA-79 mAb, which binds to a sulfation-dependent epitope. All four forms of CD34 are active in binding to E-selectin. KG1a CD34 but not spleen CD34 are active as L-selectin ligands, yet both lack MECA-79 reactivity and possess E-selectin ligand activity. This suggests that L-selectin ligands and E-selectin ligands differ in more respects than presence of the MECA-79 epitope.

%B J. Cell Biol. %V 131 %P 261-270 %G eng %N 1 %1 320 %! J. Cell Biol. %F 320 %0 Book Section %B The Harvey Lectures, Series 89 %D 1995 %T Signals on endothelium for lymphocyte recirculation and leukocyte emigration: The area code paradigm %A Springer, T.A. %B The Harvey Lectures, Series 89 %I Wiley-Liss, Inc. %C New York %P 53-103 %G eng %1 274 %F 274 %0 Journal Article %J Mol. Biol. Cell %D 1995 %T Specialized functional properties of the integrin α4 cytoplasmic domain %A Kassner,P.D. %A Alon, R. %A Springer, T.A. %A Hemler,M.E. %K CD49d %X

For functional studies of the integrin alpha 4 cytoplasmic domain, we have expressed the following in K562 and Chinese hamster ovary (CHO) cells: 1) wild-type alpha 4 (called X4C4), 2) two chimeric forms of alpha 4 (called X4C2 and X4C5) that contain the cytoplasmic domains of alpha 2 and alpha 5, respectively, and 3) alpha 4 with no cytoplasmic domain (X4C0). Cytoplasmic domain exchange had no effect on VLA-4-dependent static cell adhesion or tethering to VCAM-1 in conditions of shear flow. However, the presence of the alpha 2 or alpha 5 tails markedly enhanced VLA-4-dependent K562 cells spreading (X4C2 > X4C5 > X4C4 > X4C0), increased localization of VLA-4 into focal adhesion-like complexes in CHO cells (X4C2 > X4C5 > X4C4), and strengthened CHO and K562 cell resistance to detachment from VCAM-1 in conditions of shear flow (X4C2 > X4C5 > X4C4 > X4C0). Conversely, the alpha 4 tail supported greater VLA-4-dependent haptotactic and chemotactic cell migration. In the absence of any alpha tail (i.e., X4C0), robust focal adhesions were observed, even though cell spreading and adhesion strengthening were minimal. Thus, such focal adhesions may have relatively little functional importance, and should not be compared with focal adhesions formed when alpha tails are present. Together, these results indicate that all three alpha-chain tails exert defined positive effects (compared with no tail at all), but suggest that the alpha 4 cytoplasmic domain may be specialized to engage in weaker cytoskeletal interactions, leading to diminished focal adhesion formation, cell spreading, and adhesion strengthening, while augmenting cell migration and facilitating rolling under shear flow. These properties of the alpha 4 tail are consistent with the role of alpha 4 integrins on highly motile lymphocytes, monocytes, and eosinophils.

%B Mol. Biol. Cell %V 6 %P 661-74 %G eng %N 6 %1 279 %! Mol. Biol. Cell %F 279 %0 Book Section %B Leucocyte Typing V: White Cell Differentiation Antigens %D 1995 %T Subunit specificity and epitope mapping of Mac-1 and p150,95 mAb using chimeric CD11b x CD11c transfectants %A Luk,J. %A Luther,E. %A Diamond,M.S. %A Springer, T.A. %E Schlossman,S.F. %E Boumsell,L. %E Gilks,W. %E Harlan,J. %E Kishimoto,T. %E Morimoto,T. %E Ritz,J. %E Shaw,S. %E Silverstein,R. %E Springer, T. %E Tedder,T. %E Todd,R. %B Leucocyte Typing V: White Cell Differentiation Antigens %I Oxford University Press %C New York %P 1599-1601 %G eng %1 307 %F 307 %0 Book Section %B Atherosclerosis and coronary artery disease %D 1995 %T Traffic signals on endothelium for leukocytes in health, inflammation, and atherosclerosis %A Springer, T.A. %A Cybulsky,M.I. %E Fuster,V. %E Ross,R. %E Topol,E.J. %B Atherosclerosis and coronary artery disease %I Lippincott-Raven Publishers %C Philadelphia %P 511-537 %G eng %1 324 %F 324 %0 Journal Article %J Annu. Rev. Physiol. %D 1995 %T Traffic signals on endothelium for lymphocyte recirculation and leukocyte emigration %A Springer, T.A. %B Annu. Rev. Physiol. %V 57 %P 827-872 %G eng %1 268 %! Annu. Rev. Physiol. %F 268