Although ICAM-3 is implicated in both adhesion and signal transduction events of leukocytes, its low affinity for LFA-1 compared to other ligands of LFA-1 has puzzled many investigators. Here we investigated the role of ICAM-3 in supporting LFA-1-mediated ICAM-1 binding and subsequently cell signaling. We observed that although ICAM-3 binds poorly to LFA-1 expressed on resting T cells, it specifically facilitates and increases LFA-1-mediated adhesion to the high affinity ligand of LFA-1, ICAM-1. We demonstrate that low-affinity binding of LFA-1 to ICAM-3 together with ICAM-1 alters the cell surface distribution of LFA-1 dramatically, inducing large clusters of LFA-1 that facilitate ICAM-1 binding after LFA-1 activation. We found that LFA-1-mediated ICAM-1 cell-cell interactions such as T cell proliferation greatly depend on low affinity LFA-1/ICAM-3 interactions that enhance stable LFA-1/ICAM-1 cell-cell contact. Taken together, these data demonstrate that low affinity LFA-1 binding to ICAM-3 regulates strong LFA-1/ICAM-1-mediated adhesion by driving LFA-1 into clusters to facilitate cell-cell interactions that take place in the immune system.

REFERENCES

Acevedo
A.
,
del Pozo
M. A.
,
Arroyo
A. G.
,
Sanchez-Mateos
P.
,
Gonzales-Amaro
R.
,
Sanchez-Madrid
F.
(
1993
).
Distribution of ICAM-3 bearing cells in normal human tissues.
Am. J. Pathol
143
,
774
783
Andrew
D.
,
Shock
A.
,
Ball
E.
,
Ortlepp
S.
,
Bell
J.
,
Robinson
M.
(
1993
).
KIM185, a monoclonal antibody to CD18 which induces a change in the conformation of CD18 and promotes both LFA-1-and CR3-dependent adhesion.
Eur. J. Immunol
23
,
2217
2222
Arroyo
A. G.
,
Campanero
M. R.
,
Sanchez-Mateos
P.
,
Zapata
J. M.
,
Ursa
M. A.
,
Delpozo
M. A.
,
Sanchez-Madrid
F.
(
1994
).
Induction of tyrosine phosphorylation during ICAM-3 and LFA-1-mediated intercellular adhesion and its regulation by the CD45 tyrosine phosphatase.
J. Cell Biol
126
,
1277
1286
Binnerts
M. E.
,
van Kooyk
Y.
,
Simmons
D. L.
,
Figdor
C. G.
(
1994
).
Distinct binding of T lymphocytes to ICAM-1,-2 or-3 upon activation of LFA-1.
Eur. J. Immunol
24
,
2155
2160
Binnerts
M. E.
,
Van kooyk
Y.
,
Edwards
C. P.
,
Champe
M.
,
Presta
L.
,
Bodary
S. C.
,
Figdor
C. G.
,
Berman
P. W.
(
1996
).
Antibodies that selectively inhibit leukocyte function-associated antigen 1 binding to intercellular adhesion molecule-3 recognize a unique epitope within the CD11a I domain.
J. Biol. Chem
271
,
9962
9968
Binnerts
M. E.
,
van Kooyk
Y.
(
1999
).
How LFA-1 binds to different ligands.
Immunol. Today
20
,
240
245
Bleijs
D. A.
,
de Waal-Malefyt
R.
,
Figdor
C. G.
,
van Kooyk
Y.
(
1999
).
Co-stimulation of T cells results in distinct IL-10 and TNF-cytokine profiles dependent on binding to ICAM-1, ICAM-2 or ICAM-3.
Eur. J Immunol
29
,
2248
2258
Buckley
C. D.
,
Ferguson
E. D.
,
Littler
A. J.
,
Bossy
D.
,
Simmons
D. L.
(
1997
).
Role of ligands in the activation of LFA-1.
Eur. J. Immunol
27
,
957
962
Burridge
K.
,
Fath
K.
(
1989
).
Focal contacts: transmembrane links between the extracellular matrix and the cytoskeleton.
BioEssays
10
,
104
108
Campanero
M. R.
,
del Pozo
M. A.
,
Arroyo
A. G.
,
Sanchez-Mateos
P.
,
Hernandez-Caselles
T.
,
Craig
A.
,
Pulido
R.
,
Sanchez-Madrid
F.
(
1993
).
ICAM-3 interacts with LFA-1 and regulates the LFA-1/ICAM-1 cell adhesion pathway.
J. Cell Biol
123
,
1007
1016
Cid
M. C.
,
Esparza
J.
,
Juan
M.
,
Miralles
A.
,
Ordi
J.
,
Vilella
R.
,
Urbanomarquez
A.
,
Gaya
A.
,
Vives
J.
,
Yague
J.
(
1994
).
Signaling through CD50 (ICAM-3) stimulates T lymphocyte binding to human umbilical vein endothelial cells and extracellular matrix proteins via an increase in beta 1 and beta 2 integrin function.
Eur. J. Immunol
24
,
1377
1382
Clark
E. A.
,
Brugge
J. S.
(
1995
).
Integrins and signal transduction pathways: the road taken.
Science
268
,
233
239
Cordell
J. L.
,
Pulford
K.
,
Turley
H.
,
Jones
M.
,
Micklem
K.
,
Doussis
I. A.
,
Tyler
X.
,
Mayne
K.
,
Gatter
K. C.
,
Mason
D. Y.
(
1994
).
Cellular distribution of human leukocyte adhesion molecule ICAM-3.
J. Clin. Pathol
47
,
143
147
Craig
S. W.
,
Johnson
R. P.
(
1996
).
Assembly of focal adhesions: progress, paradigms and potents.
Curr. Opin. Cell Biol
8
,
74
85
de Fougerolles
A. R.
,
Stacker
S. A.
,
Schwarting
R.
,
Springer
T. A.
(
1991
).
Characterization of ICAM-2 and evidence for a third counter-receptor for LFA-1.
J. Exp. Med
174
,
253
267
de Fougerolles
A. R.
,
Springer
T. A.
(
1992
).
Intercellular adhesion molecule 3, a third adhesion counter-receptor for lymphocyte function-associated molecule 1 on resting lymphocytes.
J. Exp. Med
175
,
185
190
de Fougerolles
A. R.
,
Klickstein
L. B.
,
Springer
T. A.
(
1993
).
Cloning and expression of intercellular adhesion molecule 3 reveals strong homology to other immunoglobulin family counter-receptors for lymphocyte function-associated antigen 1.
J. Exp. Med
177
,
1187
1192
de Fougerolles
A. R.
,
Qin
X.
,
Springer
T. A.
(
1994
).
Characterization of the Function of Intercellular Adhesion Molecule (ICAM)-3 and Comparison with ICAM-1 and ICAM-2 in Immune Responses.
J. Exp. Med
179
,
619
629
Delpozo
M. A.
,
Campanero
M. R.
,
Sanchez-Mateos
P.
,
Arroyo
A. G.
,
Pulido
R.
,
Munoz
C.
,
Hernandez-Caselles
T.
,
Aparicio
P.
,
Sanchez-Madrid
F.
(
1994
).
Role of ICAM-3 in intercellular adhesion and activation of T lymphocytes.
Cell Adhes. Commun
2
,
211
218
Diacovo
T. G.
,
Defougerolles
A. R.
,
Bainton
D. F.
,
Springer
T. A.
(
1994
).
A functional integrin ligand on the surface of platelets: Intercellular adhesion molecule-2.
J. Clin. Invest
94
,
1243
1251
Dransfield
I.
,
Hogg
N.
(
1989
).
Regulated expression of Mg2+binding epitope on leukocyte integrin alpha subunits.
EMBO J
8
,
3759
3765
Dransfield
I.
,
Cabanas
C.
,
Craig
A.
,
Hogg
N.
(
1992
).
Divalent cation regulation of the function of the leukocyte integrin LFA-1.
J. Cell Biol
116
,
219
226
Dustin
M. L.
,
Rothlein
R.
,
Bhan
A. K.
,
Dinarello
C. A.
,
Springer
T. A.
(
1986
).
Induction by IL1 and interferon-gamma: tissue distribution, biochemistry and function of a natural adherence molecule (ICAM-1).
J. Immunol
137
,
245
254
Dustin
M. L.
,
Springer
T. A.
(
1989
).
T-cell receptor cross-linking transiently stimulates adhesiveness through LFA-1.
Nature
341
,
619
624
Fawcett
J.
,
Holness
C. L.
,
Needham
L. A.
,
Turley
H.
,
Gatter
K. C.
,
Mason
D. Y.
,
Simmons
D. L.
(
1992
).
Molecular cloning of ICAM-3, a third ligand for LFA-1, constitutively expressed on resting leukocytes.
Nature
360
,
481
484
Figdor
C. G.
,
Van Es
W. L.
,
Leemans
J. M.
,
Bont
W. S.
(
1984
).
A centrifugal elutriation system of separating small numbers of cells.
J. Immunol. Meth
68
,
73
87
Figdor
C. G.
,
van Kooyk
Y.
,
Keizer
G. D.
(
1990
).
On the mode of action of LFA-1.
Immunol. Today
11
,
277
280
Hernandez-Caselles
T.
,
Rubio
G.
,
Campanero
M. R.
,
Angel
d. P.
,
Muro
M.
,
Sanchez-Madrid
F.
,
Aparicio
P.
(
1993
).
ICAM-3, the third LFA-1 counterreceptor, is a co-stimulatory molecule for both resting and activated T lymphocytes.
Eur. J. Immunol
23
,
2799
2806
Hynes
R. O.
(
1992
).
Integrins: Versatility, modulation and signaling in cell adhesion.
Cell
69
,
11
25
Imhof
B. A.
,
Dunon
D.
(
1995
).
Leukocyte migration and adhesion.
Advan. Immunol
58
,
345
416
Juan
M.
,
Vinas
O.
,
Pinootin
M. R.
,
Places
L.
,
Martinezcaceres
E.
,
Barcelo
J. J.
,
Miralles
A.
,
Vilella
R.
,
Delafuente
M. A.
,
Vives
J.
,
Yague
J.
,
Gaya
A.
(
1994
).
CD50 (intercellular adhesion molecule 3) stimulation induces calcium mobilization and tyrosine phosphorylation through p59(fyn) and p56(lck) in Jurkat T cell line.
J. Exp. Med
179
,
1747
1756
Keizer
G. D.
,
Borst
J.
,
Figdor
C. G.
,
Spits
H.
,
Miedema
F.
,
Terhorst
C.
,
De Vries
E.
(
1985
).
Biochemical and functional characteristics of the human leukocyte membrane antigen family LFA-1, Mo-1 and p150, 95.
Eur. J. Immunol
15
,
1142
1147
Keizer
G. D.
,
Visser
W.
,
Vliem
M.
,
Figdor
C. G.
(
1988
).
A monoclonal antibody (NKI-L16) directed against a unique epitope on the alpha-chain of human leukocyte function-associated antigen 1 induces homotypic cell-cell interactions.
J. Immunol
140
,
1393
1400
Krensky
A. M.
,
Sanchez-Madrid
F.
,
Robbins
E.
,
Nagy
J. A.
,
Springer
T. A.
,
Burakoff
S. J.
(
1983
).
The functional significance, distribution and structure of LFA-1, LFA-2 and LFA-3: cell surface antigens associated with CTL-target interactions.
J. Immunol
131
,
611
616
Kucik
D. F.
,
Dustin
M. L.
,
Miller
J. M.
,
Brown
E. J.
(
1996
).
Adhesion-activating phorbol ester increases the mobility of leukocyte integrin LFA-1 in cultured lymphocytes.
J. Clin. Invest
97
,
2139
2144
Landis
R. C.
,
Bennett
R. I.
,
Hogg
N.
(
1993
).
A novel LFA-1 activation epitope maps to the I domain.
J. Cell Biol
120
,
1519
1527
Landis
R. C.
,
McDowall
A.
,
Holness
C. L. L.
,
Littler
A. J.
,
Simmons
D. L.
,
Hogg
N.
(
1994
).
Involvement of the “I” domain of LFA-1 in selective binding to ligands ICAM-1 and ICAM-3.
J. Cell Biol
126
,
529
537
Leerling
M. F.
,
Vaessen
L. M.
,
Reubsaet
C. H.
,
Weimar
W.
,
Ettekoven
H.
,
Marsman
F. R.
,
Kreeftenberg
J. G.
(
1990
).
Quality control of anti human CD3 and CD4 monoclonal antibodies.
Dev. Biol. Stand
71
,
191
200
Lub
M.
,
van Kooyk
Y.
,
Figdor
C. G.
(
1995
).
Ins and outs of LFA-1.
Immunol. Today
16
,
479
483
Lub
M.
,
van Kooyk
Y.
,
van Vliet
S. J.
,
Figdor
C. G.
(
1997
).
Dual role of the actin cytoskeleton in regulating cell adhesion mediated by the integrin lymphocyte function-associated molecule-1.
Mol. Biol. Cell
8
,
341
351
Lub
M.
,
van Vliet
S. J.
,
Oomen
S. P.
,
Pieters
R. A.
,
Robinson
M.
,
Figdor
C. G.
,
van Kooyk
Y.
(
1997
).
Cytoplasmic tails of beta 1, beta 2 and beta 7 integrins differentially regulate LFA-1 function in K562 cells.
Mol. Biol Cell
8
,
719
728
Mackay
C.
(
1995
).
Lymphocyte migration: A new spin on lymphocyte homing.
Curr. Biol
5
,
733
736
Marlin
S. D.
,
Springer
T. A.
(
1987
).
Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen 1 (LFA-1).
Cell
51
,
813
819
Martz
E.
(
1987
).
LFA-1 and other accessory molecules functioning in adhesions of T and B lymphocytes.
Hum. Immunol
18
,
3
37
Pavalko
F. M.
,
LaRoche
S. M.
(
1993
).
Activation of human neutrophils induces an interaction between the integrin beta2-subunit (CD18) and the actin binding protein alpha-actinin.
J. Immunol
151
,
3795
3807
Robinson
M. K.
,
Andrew
D.
,
Rosen
H.
,
Brown
D.
,
Ortlepp
S.
,
Stephens
P.
,
Butcher
E. C.
(
1992
).
Antibody against the Leu-CAM beta-chain (CD18) promotes both LFA-1-and CR3-dependent adhesion events.
J. Immunol
148
,
1080
1085
Rothlein
R.
,
Springer
T. A.
(
1986
).
The requirement for lymphocyte function-associated antigen 1 in homotypic leukocyte adhesion simulated by phorbol ester.
J. Exp. Med
163
,
1132
1149
Rothlein
R.
,
Dustin
M. L.
,
Marlin
S. D.
,
Springer
T. A.
(
1986
).
A human intercellular adhesion molecule (ICAM-1) distinct from LFA-1.
J. Immunol
137
,
1270
1274
Sanchez-Madrid
F.
,
Krensky
A. M.
,
Ware
C. F.
,
Robbins
E.
,
Strominger
J. L.
,
Burakoff
S. J.
,
Springer
T. A.
(
1982
).
Three distinct antigens associated with human T-lymphocyte-mediated cytolysis: LFA-1, LFA-2, and LFA-3.
Proc. Nat. Acad. Sci. USA
79
,
7489
7493
Sanchez-Madrid
F.
,
Nagy
J. A.
,
Robbins
E.
,
Simon
P.
,
Springer
T. A.
(
1983
).
A human leukocyte differentiation antigen family with distinct alpha-subunits and a common beta-subunit: the lymphocyte function-associated antigen (LFA-1), the C3bi complement receptor (OKM1/Mac-1) and the p150, 95 molecule.
J. Exp. Med
158
,
1785
1803
Simmons
D.
,
Makgoba
M. W.
,
Seed
B.
(
1988
).
ICAM, an adhesion ligand of LFA-1, is homologous to the neural cell adhesion molecule NCAM.
Nature
331
,
624
627
Spits
H.
,
Keizer
G.
,
Borst
J.
,
Terhorst
C.
,
Hekman
A.
,
de Vries
J. E.
) (
1983
).
Characterization of monoclonal antibodies against cell surface molecules associated with cytotoxic activity of natural and activated killer cells and cloned CTL lines.
Hybridoma
2
,
423
437
Springer
T. A.
(
1990
).
Adhesion receptors of the immune system.
Nature
346
,
425
434
Starling
G. C.
,
Mclellan
A. D.
,
Egner
W.
,
Sorg
R. V.
,
Fawcett
J.
,
Simmons
D. L.
,
Hart
D. N. J.
(
1995
).
Intercellular adhesion molecule-3 is the predominant co-stimulatory ligand for leukocyte function antigen-1 on human blood dendritic cells.
Eur. J. Immunol
25
,
2528
2532
Staunton
D. E.
,
Marlin
S. D.
,
Stratowa
C.
,
Dustin
M. L.
,
Springer
T. A.
(
1988
).
Primary structure of ICAM-1 demonstrates interaction between members of the immunoglobulin and integrin supergene families.
Cell
52
,
925
933
Staunton
D. E.
,
Dustin
M. L.
,
Springer
T. A.
(
1989
).
Functional cloning of ICAM-2, a cell adhesion ligand for LFA-1 homologous to ICAM-1.
Nature
339
,
61
64
Stewart
M.
,
Hogg
N.
(
1996
).
Regulation of leukocyte integrin function: Affinity vs avidity.
J. Cell Biochem
61
,
554
561
van Kooyk
Y.
,
van de Wiel-van Kemenade
P.
,
Weder
P.
,
Kuijpers
T. W.
,
Figdor
C. G.
(
1989
).
Enhancement of LFA-1-mediated cell adhesion by triggering through CD2 or CD3 on T lymphocytes.
Nature
342
,
811
813
van Kooyk
Y.
,
Weder
P.
,
Hogervorst
F.
,
Verhoeven
A. J.
,
van Seventer
G.
,
te
V.
,
Borst
J.
,
Keizer
G. D.
,
Figdor
C. G.
(
1991
).
Activation of LFA-1 through a Ca2(+)-dependent epitope stimulates lymphocyte adhesion.
J. Cell Biol
112
,
345
354
van Kooyk
Y.
,
Figdor
C. G.
(
1993
).
Lymphocyte adhesion mediated by integrins.
Res. Immunol
144
,
709
722
van Kooyk
Y.
,
van de Wiel-van Kemenade
E.
,
Weder
P.
,
Huijbens
R. J. F.
,
Figdor
C. G.
(
1993
).
Lymphocyte function-associated antigen 1 dominates very late antigen 4 of activated T cells to endothelium.
J. Exp. Med
177
,
185
190
van Kooyk
Y.
,
Weder
P.
,
Heije
K.
,
Figdor
C. G.
(
1994
).
Extracellular Ca2+ modulates leukocyte function-associated antigen-1 cell surfacedistribution on T lymphocytes and consequently affects cell adhesion.
J. Cell Biol
124
,
1061
1070
van Kooyk
Y.
,
Binnerts
M. E.
,
Edwards
C. P.
,
Champe
M.
,
Berman
P. W.
,
Figdor
C. G.
,
Bodary
S. C.
(
1996
).
Critical amino acids in the lymphocyte function-associated antigen-1 I domain mediate intercellular adhesion molecule 3 binding and immune function.
J. Exp. Med
183
,
1247
1252
Vandervieren
M.
,
Letrong
H.
,
Wood
C. L.
,
Moore
P. F.
,
Stjohn
T.
,
Staunton
D. E.
,
Gallatin
W. M.
(
1995
).
A novel leukointegrin, alpha d beta 2, binds preferentially to ICAM-3.
Immunity
3
,
683
690
Vazeux
R.
,
Hoffman
P. A.
,
Tomita
J. K.
,
Dickinson
E. S.
,
Jasman
R. L.
,
Stjohn
T.
,
Gallatin
W. M.
(
1992
).
Cloning and characterization of a new intercellular adhesion molecule ICAM-R.
Nature
360
,
485
488
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