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Science 18 December 1998: Vol. 282. no. 5397, pp. 2266 - 2269 DOI: 10.1126/science.282.5397.2266
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Reports
A Receptor/Cytoskeletal Movement Triggered by Costimulation During T Cell Activation
Christoph Wülfing,
Mark M. Davis
*
During T cell activation, the engagement of costimulatory molecules
is often crucial to the development of an effective immune response,
but the mechanism by which this is achieved is not known. Here, it is
shown that beads attached to the surface of a T cell translocate toward
the interface shortly after the start of T cell activation. This
movement appears to depend on myosin motor proteins and requires the
engagement of the major costimulatory receptor pairs, B7-CD28 and
ICAM-1-LFA-1. This suggests that the engagement of costimulatory
receptors triggers an active accumulation of molecules at the interface
of the T cell and the antigen-presenting cell, which then increases the
overall amplitude and duration of T cell signaling.
Howard Hughes Medical Institute and Department of Microbiology and
Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
*
To whom correspondence should be addressed.
The central event in T cell
activation is the interaction of the T cell receptor (TCR) with the
antigenic peptide presented by the major histocompatibility complex
(MHC) of the antigen-presenting cell (APC). However, because the number
of agonist peptide-MHC complexes can be very low, in the range of 10 to 100 per APC (1), and because the TCR is continuously
modulated from the T cell surface (2), sustained T cell
activation is likely to require signal amplification (3,
4). An important component of this amplification is thought
to be provided by costimulatory molecules on the T cell, although the
mechanism by which they accomplish it is unclear (5).
The most important of the costimulatory receptors on T cells and their
ligand on APCs are CD28-B7 (6) and LFA-1-ICAM-1
(7). Many different receptor couples, including
TCR-peptide-MHC and LFA-1-ICAM-1, accumulate at the T cell-APC
interface (8, 9). This accumulation has been assumed to be a
passive, diffusion-limited cocapping mechanism (10). Here,
we describe an active, cytoskeletal mechanism that appears to drive
receptor accumulation at the T cell-APC interface. This mechanism
requires the APC to express B7 and ICAM-1 and is independent of TCR
signaling. We suggest that this mechanism is a central part of
costimulation, as it would effectively amplify any TCR-mediated
signals.
To study whether receptor accumulation at the T cell-APC interface
could be actively driven by the T cell cytoskeleton, we monitored the
general movement of the cortical actin cytoskeleton and linked
receptors (11) using the classical technique of attaching
large beads to the surfaces of antigen-specific T cells. We
coated 4.5-µm beads with an antibody to the T cell surface antigen
ICAM-1 (12). Cross-linking ICAM-1 by beads in this way is
not expected to influence T cell function (13). As has been
observed with fibroblasts (11, 14), we find that in
migrating 5C.C7 transgenic T cells (9, 15) the beads
translocate from the anterior to the posterior end of the cell
(16). We then mixed bead-loaded 5C.C7 T cells
with B cell lymphoma cells that express the appropriate MHC molecule
(I-Ek) and have been pulsed with the moth cytochrome c
peptide 88-103. After contact with the APC, the T cells rapidly become
activated (9, 17) and the beads move from the
posterior end of the T cell to the newly formed interface with the B
cells (Fig. 1, movie 1) beginning 4 ± 1 min after the first rise in intracellular calcium
(n = 13). This suggests that the T cell cortical actin
cytoskeleton reorients toward the T cell-APC interface soon after the
start of T cell activation. The ensuing cytoskeletal flow would allow
receptors that are linked to the actin cytoskeleton to be transported
to the newly formed T cell-APC interface.
Fig. 1.
Bead movement toward the
interface. Single frames of a video microscopy experiment
(36) of the interaction of 5C.C7 T cells, loaded with
4.5-µm anti-ICAM-1 (YN1) beads, with peptide-loaded CH27 B
cell lymphoma cells (35) are shown. The CH27 cell is
substantially larger than the T cells. The T cell intracellular calcium
concentration is overlaid in a false color scale from blue (low
concentration) to red (high concentration) to mark the onset of T cell
activation, set to time 0:00 min. Although the beads, one of
which is marked with an arrow, are bound to the posterior end of the
central T cell before and at the time of its activation, they can be
seen to move toward the T cell-B cell interface in subsequent frames.
The still frames have been excerpted from movie 1 (37),
which can be viewed at Science Online
(http://www.sciencemag.org/feature/data/984937.shl).
[View Larger Version of this Image (80K GIF file)]
To rule out ICAM-1-specific effects, we also attached beads to T
cells in two other ways. First, we surface-biotinylated the 5C.C7
T cells with an amine-reactive form of biotin and used 2.8-µm streptavidin beads (18). Second, we exchanged a lipid that
is biotinylated at its head into the cell surface of the T cells from
liposomes (19) and used the same 2.8-µm streptavidin beads
(20). In both cases, we found a behavior identical to that
of the antibody-coated beads (Table 1). This confirms that the beads monitor the movement of the cortical actin
cytoskeleton and not that of a specific receptor. We have also obtained
identical results using monocytes and dendritic cells prepared directly
ex vivo as APCs (16). The time to travel half a cell
circumference is 6 ± 2 min (n = 14). Given that
the diameter of our activated T cells is ~10 µm, this speed of ~3
µm/min is similar to that reported in comparable studies on migrating
fibroblasts (1 to 5 µm/min) (21). This behavior is
independent of the peptide concentration on the APC as long as enough
peptide is presented to activate the T cell (16).
Beads bound to the B cell do not move, consistent with the lack of
involvement of the B cell actin cytoskeleton in the redistribution of
specific receptors (9, 22).
Table 1.
Occurrence of different types of T cell bead movement
(39) during the interaction of a 5C.C7 T cell with APCs
loaded with an activating peptide. Unless mentioned, the APCs were CH27
B cell lymphomas. Bead movement was analyzed using three different
types of beads (12, 18, 20); only two of these bead types
(12, 18) were used for the rest of the
experiments. Blocking antibodies and pharmacological agents were used
under standard conditions. (40). The number of cells
analyzed during wortmannin treatment was small because the majority of
the T cells did not establish a polarized phenotype (41).
When transfected CHO cells were used as APCs, we found that GFP fusion
did not inhibit the function of its fusion partner (9, 22).
In the last row, an activating antibody to CD28 was present during the
interaction of the T cell and APC; n indicates the number of
cells studied
(34).
|
| Type of
beads/treatment/APC |
Movement toward interface |
Movement away from
interface |
No movement |
n |
|
| Type of
beads |
| Anti-ICAM |
74% |
0% |
26% |
81 |
| Streptavidin
(surface biotin) |
83% |
7% |
10% |
29 |
| Streptavidin (lipid
biotin) |
64% |
5% |
32% |
22 |
| Antibody
blocking |
| anti-ICAM |
11% |
5% |
84% |
37 |
| anti-B7 |
8% |
0% |
92% |
24 |
| anti-VCAM |
62% |
0% |
38% |
21 |
| anti-CD48 |
75% |
0% |
25% |
20 |
| Pharmacological
agents |
| Wortmannin |
13% |
0% |
78% |
8 |
| Ni/BAPTA |
23% |
0% |
77% |
26 |
| Transfected
CHO cells as
APCs |
| CHO/I-Ek |
4% |
0% |
96% |
47 |
| CHO/I-Ek/ICAM-1-GFP |
27% |
7% |
67% |
45 |
| CHO/I-Ek/B7-2-GFP |
34% |
2% |
64% |
47 |
| CHO/I-Ek/ICAM-1-GFP
(anti-CD28) |
59% |
0% |
41% |
27 |
|
To study the role of costimulatory receptor pairs in the regulation of
the cytoskeletal movement, we added antibodies that block a number of
accessory interactions to this system. We found that blocking ICAM-1 or
B7-1 together with B7-2 abolishes bead movement while leaving
undisturbed other aspects of T cell activation, such as the formation
of a tight interface and the elevation of intracellular calcium (Table
1) (16). Antibodies to CD48 or VCAM had no effect
(Table 1). To determine whether the interactions of ICAM-1 with LFA-1
or of B7 with CD28 are sufficient to trigger bead movement, we used
transfected CHO cells as APCs. When transfected with
I-Ek, the CHO cells were not able to induce bead movement
(Table 1). When the CHO cells were transfected with I-Ek
and either ICAM-1 or B7-2 gene constructs, we observed a partial restoration of the bead movement (Table 1). The best results were
obtained by transfecting ICAM-1 and engaging CD28 on the T cell with an
activating antibody, which allows bead movement to near B cell levels
(Table 1). Preliminary experiments with CHO cells expressing all three
molecules (I-Ek, ICAM-1, and B7-2) showed that 7 of 10 T-CHO couples examined moved beads toward the T cell-APC interface
(16). These experiments show that the interactions
of LFA-1-ICAM-1 and CD28-B7 are both necessary and sufficient to
induce movement of the T cell cortical actin cytoskeleton toward the
newly formed T cell-APC interface.
We have also characterized second messengers that are
synergistically activated by the TCR and the accessory
interactions that are important for bead movement, namely the
intracellular calcium concentration for ICAM-1-LFA-1 (9,
23) and the phosphatidylinositol 3-kinase (PI 3-kinase) for
B7-CD28 (24). Blocking the rise in the intracellular
calcium concentration with
1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) and Ni2+ blocks bead movement (Table 1)
while leaving the formation of a tight interface intact (9).
Using the PI 3-kinase inhibitor wortmannin (25) at
100 nM leads to a distinct phenotype in that most T cells do not
establish a polarized phenotype before activation. For the minority of
T cells that display a polarized phenotype, bead movement is abolished.
These results are consistent with both PI 3-kinase and
intracellular calcium being necessary for bead movement.
The cortical actin cytoskeleton moves either by controlled assembly and
disassembly of actin filaments or by sliding actin filaments past each
other using myosin motor proteins (11, 26). In
particular, myosin motor proteins are thought to be involved in actin
cytoskeleton movement at the cell body (21). Previously, we
and others have shown that blocking actin filament assembly in the T
cell (but not B cell APCs) with cytochalasin D severely disrupts many
aspects of T cell activation (3, 9). Inhibiting the myosin
motors with butanedione monoxime (BDM) (27) gives a
more specific phenotype in this system, as shown in Fig.
2 (movie 2) and Table
2. At high concentrations of BDM, the
diameter of the T cell-B cell interface is reduced, the calcium signal is less sustained, the redistribution of ICAM-1 to the T cell-B cell
interface is impaired, the redistribution of I-Ek is
inhibited (22), and bead movement is partially blocked
(Table 2). At low BDM concentrations (2 mM), the reduction in interface
diameter and the impairment of bead movement remain (Table 2),
suggesting that myosin motors are most involved in these two processes.
Because BDM has also been reported to block T cell potassium uptake
(28), we inhibited voltage-gated potassium channels
specifically with noxius toxin (29) and found no inhibition
of bead movement (Table 2). The small effect we see on early T cell
activation is consistent with the moderate effects reported for
specific potassium channel blockers on later activation events
(30). We conclude that the receptor/cytoskeletal movement
that we describe here is dependent on myosin motor proteins.
Fig. 2.
T cell-APC interaction with
blocked myosin motor proteins. (A) Single frames of a video
microscopy experiment (36) of the interaction of 5C.C7 T
cells, loaded with 4.5-µm anti-ICAM-1 (YN1) beads, with
peptide-loaded, ICAM-1-GFP-transfected CH27 cells are shown. The
bright-field image has been duplicated. In the top panels, the T cell
intracellular calcium concentration is overlaid in a false color scale
as in Fig. 1. In the bottom panels, the ICAM-1-GFP fluorescence is
overlaid in a false color scale from green (low fluorescence) to blue
(high fluorescence) to allow the simultaneous investigation of the
additional cellular parameter. The accumulation of ICAM-1 at the T
cell-APC interface in the absence of all pharmacological agents was
described in (9); here, such an interaction is shown in the
presence of 2 µM BDM. Although the calcium signal is elevated in a
stable manner and ICAM-1-GFP accumulates at the interface, this
interface is narrow (as most easily seen by the tightly concentrated
ICAM-1-GFP accumulation), and a bead that is bound to an activating T
cell, marked with an arrow, does not move. The still frames were
excerpted from movie 2 (38), which can be viewed at
Science Online (http://www.sciencemag.org/feature/data/984937.shl). (B) An even narrower interface at 20 µM BDM, excerpted from a movie of a different experiment.
[View Larger Version of this Image (54K GIF file)]
Table 2.
Effects of blocking myosin motor proteins. The
analysis of T cell activation by peptide loaded CH27 cells in the
presence of BDM or noxius toxin (NTX) is shown (42).
Narrow interface form denotes activated T cells having a T cell-B cell
interface diameter that is smaller than the diameter of the T cell
(Fig. 2). Calcium signal back to baseline denotes activated T cells
whose intracellular calcium concentration returns to preactivation
levels during an observation period of at least 5 min. ICAM cluster
formation denotes CH27 cells that show accumulation of ICAM-1-GFP at
the T cell-B cell interface after T cell activation (9)
(Fig. 2). Bead movement toward the interface (39) refers to
activated T cells that show anti-ICAM-1 bead movement. In all cases,
n denotes the number of cells analyzed
(43).
|
| Pharmacol.
agent |
Interface form
|
Calcium
signal
|
ICAM clustering
|
Bead
movement
|
| Narrow |
n |
Back to baseline |
n |
Cluster
formation |
n |
Toward interface |
n |
|
| BDM (20 mM) |
93% |
55 |
68% |
108 |
36% |
33 |
19% |
27 |
| BDM (10 mM) |
79% |
58 |
45% |
83 |
51% |
35 |
27% |
15 |
| BDM (2 mM) |
70% |
46 |
23% |
73 |
85% |
32 |
25% |
28 |
| Buffer
alone |
14% |
56 |
2% |
102 |
90% |
38 |
62% |
21 |
| NTX
(100 nM) |
23% |
47 |
9% |
76 |
85% |
40 |
63% |
27 |
|
In vivo, costimulation is usually required for efficient T cell
activation, probably because TCR signaling is limited by small numbers
of agonist peptide-MHC complexes and continuous internalization of the
TCR-CD3 complex. Previously, we have shown that the accumulation of
MHC-peptide-TCR and accessory receptor couples at the T cell-APC interface after T cell activation can amplify the weak TCR signal efficiently (9, 22). Here, we have described an active
movement of the cortical actin cytoskeleton toward the interface,
involving myosin motor proteins, that would move all molecules linked
to the actin cytoskeleton toward the T cell-APC interface. In support
of this possibility, we find that the increase in concentration of
LFA-1-ICAM-1 and other (TCR-peptide-I-Ek and CD2-CD48)
receptor couples at the interface and the movement of the cortical
cytoskeleton are regulated similarly. Both processes are sensitive to
the myosin inhibitor BDM, to anti-B7, and to the choice of APC
(9, 22) (Table 2). Furthermore, we have shown that this
movement of the actin cytoskeleton is regulated by the two most
important costimulatory receptor pairs, CD28-B7 and LFA-1-ICAM-1. We
suggest that this active accumulation of receptor pairs and other
cytoskeleton-linked molecules at the T cell-APC interface, and the
signal amplification that would result from these increased receptor
densities, could be the principal basis of the costimulatory effect.
This is in contrast to models in which a costimulatory signal
integrates with TCR signals in the nucleus to affect gene expression.
REFERENCES AND NOTES
-
S. Demotz,
H. M. Grey,
A. Sette,
Science
249,
1028
(1990)
[Abstract/Free Full Text]
;
C. V. Harding and
E. R. Unanue,
Nature
346,
574
(1990)
[CrossRef] [Medline]
;
R. C. Brower,
et al.,
Mol. Immunol.
31,
1285
(1994)
[CrossRef] [Web of Science] [Medline]
;
Y. Sykulev,
M. Joo,
I. Vturina,
T. J. Tsomides,
H. N. Eisen,
Immunity
4,
565
(1996)
[CrossRef] [Web of Science] [Medline]
;
J. Delon,
N. Bercovici,
G. Raposo,
R. Liblau,
A. Trautmann,
J. Exp. Med.
188,
1473
(1998)
[Abstract/Free Full Text]
.
-
S. Valitutti,
S. Müller,
M. Cella,
E. Padovan,
A. Lanzavecchia,
Nature
375,
148
(1995)
[CrossRef] [Medline]
;
S. Valitutti,
S. Müller,
M. Salio,
A. Lanzavecchia,
J. Exp. Med.
185,
1859
(1997)
[Abstract/Free Full Text]
.
-
S. Valitutti,
M. Dessing,
K. Aktories,
H. Gallati,
A. Lanzavecchia,
J. Exp. Med.
181,
577
(1995)
[Abstract/Free Full Text]
.
-
G. Iezzi,
K. Karjalainen,
A. Lanzavecchia,
Immunity
8,
89
(1998)
[CrossRef] [Web of Science] [Medline]
;
L. J. Holsinger,
et al.,
Curr. Biol.
8,
563
(1998)
[CrossRef] [Web of Science] [Medline]
.
-
C. G. Sagerstrom,
E. M. Kerr,
J. P. Allison,
M. M. Davis,
Proc. Natl. Acad. Sci. U.S.A.
90,
8987
(1993)
[Abstract/Free Full Text]
;
M. Croft and
C. Dubey,
Crit. Rev. Immunol.
17,
89
(1997)
[Web of Science] [Medline]
;
S. L. Swain,
et al.,
Immunol. Rev.
150,
143
(1996)
[CrossRef] [Web of Science] [Medline]
;
M. F. Bachmann,
et al.,
Immunity
7,
549
(1997)
[CrossRef] [Web of Science] [Medline]
.
-
C. A. Chambers and
J. P. Allison,
Curr. Opin. Immunol.
9,
396
(1997)
[CrossRef] [Web of Science] [Medline]
;
P. J. Blair,
et al.,
Biochem. Soc. Trans.
25,
651
(1997)
[Web of Science] [Medline]
;
A. I. Sperling and
J. A. Bluestone,
Immunol. Rev.
153,
155
(1996)
[CrossRef] [Web of Science] [Medline]
.
-
M. L. Dustin and
T. A. Springer,
Nature
341,
619
(1989)
[CrossRef] [Medline]
;
Y. van Kooyk and
C. G. Figdor,
Biochem. Soc. Trans.
25,
515
(1997)
[Web of Science] [Medline]
;
M. Stewart and
N. Hogg,
J. Cell. Biochem.
61,
554
(1996)
[CrossRef] [Web of Science] [Medline]
.
-
A. Kupfer and
S. J. Singer,
Annu. Rev. Immunol.
7,
309
(1989)
[CrossRef] [Web of Science] [Medline]
;
C. R. Monks,
B. A. Freiberg,
H. Kupfer,
N. Sciaky,
A. Kupfer,
Nature
395,
82
(1998)
[CrossRef] [Medline]
.
-
C. Wülfing,
M. D. Sjaastad,
M. M. Davis,
Proc. Natl. Acad. Sci. U.S.A.
95,
6302
(1998)
[Abstract/Free Full Text]
.
-
S. J. Singer,
Science
255,
1671
(1992)
[Abstract/Free Full Text]
.
-
As reviewed by
M. D. Sheets,
R. Simson,
K. Jacobson,
Curr. Opin. Cell Biol.
7,
707
(1995)
[CrossRef] [Web of Science] [Medline]
;
M. S. Bretscher,
Cell
87,
601
(1996)
[CrossRef] [Web of Science] [Medline]
.
-
Dynabeads (M450; Dynal) coated with sheep antisera to rat
immunoglobulin were coated with YN1 (31) at 4°C at 160 µg per 107 beads. For microscopy, beads were used at a
2:1 ratio over T cells. Half of the beads were preincubated
with the T cells at 4°C; the other half were added to the T cells on
the microscopy stage.
-
H. Xu,
et al.,
J. Exp. Med.
180,
95
(1994)
[Abstract/Free Full Text]
;
J. E. Sligh Jr.,
et al.,
Proc. Natl. Acad. Sci. U.S.A.
90,
8529
(1993)
[Abstract/Free Full Text]
.
-
B. F. Holifield,
A. Ishihara,
K. Jacobson,
J. Cell Biol.
111,
2499
(1990)
[Abstract/Free Full Text]
;
D. A. Lauffenberger and
A. F. Horwitz,
Cell
84,
359
(1996)
[CrossRef] [Web of Science] [Medline]
.
-
R. A. Seder, W. E. Paul, M. M. Davis, B. Fazekas de St. Groth, J. Exp. Med. 176, 1091 (1992).
-
C. Wülfing and M. M. Davis, data not shown.
-
P. A. Negulescu,
T. B. Krasieva,
A. Khan,
H. H. Kerschbaum,
M. D. Cahalan,
Immunity
4,
421
(1996)
[CrossRef] [Web of Science] [Medline]
.
-
T cells were surface-biotinylated after loading them with
Fura-2 by incubating 106 T cells for 30 min at 4°C in
Ringer's buffer [10 mM Hepes (pH 7.4), 154 mM NaCl, 7.2 mM KCl, and
1.8 mM CaCl2] with sulfo-NHS-biotin (200 µg/ml; Pierce,
Rockford, IL). After washing the cells into 10% fetal bovine serum in
phosphate-buffered saline (PBS) with 1 mM CaCl2 and 0.5 mM
MgCl2 at 4°C, they were used within 2 hours.
Streptavidin-coated Dynabeads (M280; Dynal) were used as described for
the YN1-coated beads.
-
A. E. Ting and
R. E. Pagano,
J. Biol. Chem.
265,
5337
(1990)
[Abstract/Free Full Text]
.
-
Biotinylated lipids were incorporated into the T cell
membrane from liposomes consisting of 85% egg phosphatidylcholine
(Sigma), 10%
N-([6-(biotinoyl)amino]hexanoyl)1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine
(Molecular Probes), and 5%
N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine
(Molecular Probes), prepared as described (32). The
biotinylated lipids were incorporated into the T cell
membrane by incubating 106 T cells for 1 hour with the
liposomes at a lipid concentration of 0.1 mg/ml in PBS with 1 mM
CaCl2 and 0.5 mM MgCl2.
-
J. P. Heath and
B. F. Holifield,
Symp. Soc. Exp. Biol.
47,
35
(1993)
[Medline]
.
-
C. Wülfing and M. M. Davis, unpublished data.
-
P. Kuhlman,
V. T. Moy,
B. A. Lollo,
A. A. Brian,
J. Immunol.
146,
1773
(1991)
[Abstract]
;
S. B. Kanner,
L. S. Grosmaire,
J. A. Ledbetter,
N. K. Damle,
Proc. Natl. Acad. Sci. U.S.A.
90,
7099
(1993)
[Abstract/Free Full Text]
.
-
S. G. Ward,
C. H. June,
D. Olive,
Immunol. Today
17,
187
(1996)
[CrossRef] [Web of Science] [Medline]
;
A. Toker and
L. C. Cantley,
Nature
387,
673
(1997)
[CrossRef] [Medline]
.
-
H. Yano,
et al.,
J. Biol. Chem.
268,
25846
(1993)
[Abstract/Free Full Text]
;
R. Woscholski,
T. Kodaki,
M. McKinnon,
M. D. Waterfield,
P. J. Parker,
FEBS Lett.
342,
109
(1994)
[CrossRef] [Web of Science] [Medline]
.
-
As reviewed by
M. F. Carlier and
D. Pantaloni,
J. Mol. Biol.
269,
459
(1997)
[CrossRef] [Web of Science] [Medline]
;
S. K. Maciver,
Bioessays
18,
179
(1996)
[CrossRef] [Web of Science] [Medline]
.
-
L. P. Cramer and
T. J. Mitchison,
J. Cell Biol.
131,
179
(1995)
[Abstract/Free Full Text]
.
-
L. C. Schlichter,
P. A. Pahapill,
I. Chung,
J. Pharmacol. Exp. Ther.
261,
438
(1992)
[Abstract/Free Full Text]
.
-
S. Grissmer,
et al.,
Mol. Pharmacol.
45,
1227
(1994)
[Abstract]
.
-
J. A. Verheugen,
F. Le Deist,
V. Devignot,
H. Korn,
Cell Calcium
21,
1
(1997)
[CrossRef] [Web of Science] [Medline]
and references therein.
-
F. Takei,
J. Immunol.
134,
1403
(1985)
[Abstract]
.
-
J. T. Groves,
C. Wülfing,
S. G. Boxer,
Biophys. J.
71,
2716
(1996)
[Web of Science] [Medline]
.
-
K. Miyake,
et al.,
J. Cell Biol.
114,
557
(1991)
[Abstract/Free Full Text]
;
K. Miyake,
I. L. Weissman,
J. S. Greenberger,
P. W. Kincade,
J. Exp. Med.
173,
599
(1991)
[Abstract/Free Full Text]
.
-
The results are derived from a composite analysis of at least
seven experiments (11 on average) performed on at least two different
days (four on average) for each condition. We show a composite
analysis because on average only three or four cells per experiment
could be analyzed. To ensure consistency within the data set, at least
one positive control experiment was included during each day of
experiments.
-
The CH27 cells used in this particular experiment were
ICAM-1-GFP transfected. ICAM-1-GFP transfection of the CH27 cells has
no effect on the bead movement (16). The ICAM-1-GFP
fluorescence is not shown.
-
Peptide loading of APCs using 10 µM moth cytochrome c
peptide 82-103, Fura-2 loading of T cells, and microscopy (using the
Zeiss/Attofluor system) were performed as in (9).
-
Movie 1: Bead movement toward the interface. The interaction
of 5C.C7 T cells, loaded with 4.5-µm anti-ICAM-1 (YN1) beads,
with peptide-loaded CH27 B cell lymphoma cells (35) is
shown. The CH27 cell is substantially larger than the T cells. Although
the beads are bound to the posterior end of the top T cell before and
at the time of its activation, they can be seen to move toward the T
cell-B cell interface in subsequent frames. The T cell intracellular
calcium concentration is overlaid in a false color scale. Two
closely related calcium-sensitive dyes that differ in their
calcium dissociation constant, Fura-PE in movie 1 and Fura-2 in movie
2, have been used. Therefore, blue indicates low calcium concentration
in both movies, whereas high calcium concentration is encoded in yellow
in movie 1 and in red in movie 2. For reasons of simplicity, in Fig. 1
the calcium color scale has been changed from the original one of movie
1 to match that of movie 2/Fig. 2. The movie compresses 20 min of
experiment into 1 min of movie.
-
Movie 2: T cell-APC interaction with blocked myosin motor
proteins. The interaction of 5C.C7 T cells, loaded with 4.5-µm
anti-ICAM-1 (YN1) beads, with peptide-loaded,
ICAM-1-GFP-transfected CH27 B cell lymphoma in the presence of 2 mM
BDM is shown. A bright-field series of images has been duplicated and
is overlaid with false-color encoded fluorescence information. The top
panel is overlaid with a false-color representation of the
intracellular calcium concentration of the T cell, ranging from blue
(low concentration) to red (high concentration). In the bottom panel,
the ICAM-1-GFP fluorescence of the B cell lymphoma is overlaid in a
false color scale from green (low fluorescence) to blue (high
fluorescence) to allow the simultaneous investigation of an additional
cellular parameter. The movie shows that in the presence of 2 µM BDM,
although the calcium signal is elevated in a stable manner and
ICAM-1-GFP accumulates at the T cell-B cell interface, this interface
is narrow (as most easily seen by the tightly concentrated ICAM-1-GFP
accumulation) and no bead movement is seen. The movie compresses 20 min
of experiment into 1 min of movie.
-
Three categories of bead movement were observed.
Movement toward the interface was defined as bead movement at least
one-fourth of a T cell circumference toward the interface (as shown in
Fig. 1). Movement away from the interface was defined as bead movement
at least one-fourth of a T cell circumference away from the interface,
indicative of random bead movement. Beads located at the interface at
the moment of T cell activation constituted <25% of all beads bound
to T cells, consistent with the observation that migrating T cells move
the beads toward their posterior. The only exception occurs after
treatment with the PI 3-kinase inhibitor wortmannin, when 75% of the
beads are already at the interface at the time of T cell activation,
indicative of a lack of polarization. In this case, the beads are
likely to initiate the cell-cell interaction by binding to ICAM-1 on
both cells simultaneously. Beads that started at the interface stay
there without exception. Beads that are scored "no movement" are
those that move less than one-fourth of a T cell circumference in at
least 5 min (Fig. 2).
-
Blocking antibodies were used at 10 µg/ml. We used YN1 as
anti-ICAM-1 (31), 16-10A1 (Pharmingen) as anti-B7-1, GL1
(Pharmingen) as anti-B7-2, HM48-1 (Pharmingen) as anti-CD48, 37.51 (Pharmingen) as anti-CD28, and MK2.7 as anti-VCAM (33).
Wortmannin was used at 100 nM, Ni2+ at 5 mM, and both were
added directly to the microscopy dish. T cells were preloaded with
BAPTA in parallel with Fura-2 at 20 µM. The ICAM-1-GFP construct has
been described (9); the B7-2-GFP construct is strictly
analogous, with the GFP attached to the cytoplasmic tail. Stable
transfectants were selected for GFP expression using flow cytometry,
and single cells were cloned.
-
Lack of T cell polarization is indicated by the fact that few
activated cells had beads at the interface, by the lack of uropods and
extended lamellipodia, and by the failure to migrate (16).
-
BDM (Sigma) was added to the T cells at the indicated
concentrations 5 min before the start of the microscopy experiment from
a fresh 200 µM stock in PBS. Noxius toxin (Alomone Labs, Jerusalem,
Israel) was added to the T cells at 100 nM, that is, a 100-fold excess
over the apparent dissociation concentration of blocking peak potassium
channels 5 min before the start of the microscopy experiment.
-
The results were derived from a composite analysis of at least
five experiments (nine on average) performed on at least three
different days for each condition. As discussed in (34),
composite data sets are shown because of the small number of cells per
experiment that could be analyzed for some parameters. Positive
controls were run on each day of the experiments to ensure uniformity.
-
We thank M. D. Sjaastad, W. J. Nelson, R. S. Lewis, and D. A. Lauffenberger for helpful discussions. Supported
by the Howard Hughes Medical Institute and the European Molecular
Biology Organization (C.W.).
31 August 1998; accepted 10 November
1998
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| PDF »
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| Full Text »
| PDF »
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- C. Sumen, M. L. Dustin, and M. M. Davis (2004)
J. Cell Biol.
166, 579-590
| Abstract »
| Full Text »
| PDF »
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J. Immunol.
173, 770-775
| Abstract »
| Full Text »
| PDF »
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- J. P. O'Keefe, K. Blaine, M.-L. Alegre, and T. F. Gajewski (2004)
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| Abstract »
| Full Text »
| PDF »
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- M. V. Maus, B. Kovacs, W. W. Kwok, G. T. Nepom, K. Schlienger, J. L. Riley, D. Allman, T. H. Finkel, and C. H. June (2004)
J. Immunol.
172, 6675-6683
| Abstract »
| Full Text »
| PDF »
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- P. G. Andres, K. C. Howland, D. Dresnek, S. Edmondson, A. K. Abbas, and M. F. Krummel (2004)
J. Immunol.
172, 5880-5886
| Abstract »
| Full Text »
| PDF »
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- B. Marinari, A. Costanzo, V. Marzano, E. Piccolella, and L. Tuosto (2004)
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| Abstract »
| Full Text »
| PDF »
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- S. Le Bras, I. Foucault, A. Foussat, C. Brignone, O. Acuto, and M. Deckert (2004)
J. Biol. Chem.
279, 15550-15560
| Abstract »
| Full Text »
| PDF »
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- O. Umland, H. Heine, M. Miehe, K. Marienfeld, K. H. Staubach, and A. J. Ulmer (2004)
J. Leukoc. Biol.
75, 671-679
| Abstract »
| Full Text »
| PDF »
- PKC{theta} Signals Activation versus Tolerance In Vivo.
- N. N. Berg-Brown, M. A. Gronski, R. G. Jones, A. R. Elford, E. K. Deenick, B. Odermatt, D. R. Littman, and P. S. Ohashi (2004)
J. Exp. Med.
199, 743-752
| Abstract »
| Full Text »
| PDF »
- Cofilin peptide homologs interfere with immunological synapse formation and T cell activation.
- S. M. Eibert, K.-H. Lee, R. Pipkorn, U. Sester, G. H. Wabnitz, T. Giese, S. C. Meuer, and Y. Samstag (2004)
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101, 1957-1962
| Abstract »
| Full Text »
| PDF »
- Actin Cytoskeleton Regulates Calcium Dynamics and NFAT Nuclear Duration.
- F. V. Rivas, J. P. O'Keefe, M.-L. Alegre, and T. F. Gajewski (2004)
Mol. Cell. Biol.
24, 1628-1639
| Abstract »
| Full Text »
| PDF »
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- S. Kobayashi, K. Ohnuma, M. Uchiyama, K. Iino, S. Iwata, N. H. Dang, and C. Morimoto (2004)
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103, 1002-1010
| Abstract »
| Full Text »
| PDF »
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- K. Badour, J. Zhang, F. Shi, Y. Leng, M. Collins, and K. A. Siminovitch (2004)
J. Exp. Med.
199, 99-112
| Abstract »
| Full Text »
| PDF »
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- A. Clatza, L. C. Bonifaz, D. A. A. Vignali, and J. Moreno (2003)
J. Immunol.
171, 6478-6487
| Abstract »
| Full Text »
| PDF »
- Leishmania-Induced Inhibition of Macrophage Antigen Presentation Analyzed at the Single-Cell Level.
- C. L. Meier, M. Svensson, and P. M. Kaye (2003)
J. Immunol.
171, 6706-6713
| Abstract »
| Full Text »
| PDF »
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- K.-H. Lee, A. R. Dinner, C. Tu, G. Campi, S. Raychaudhuri, R. Varma, T. N. Sims, W. R. Burack, H. Wu, J. Wang, et al. (2003)
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302, 1218-1222
| Abstract »
| Full Text »
| PDF »
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- M. M. Al-Alwan, R. S. Liwski, S. M. M. Haeryfar, W. H. Baldridge, D. W. Hoskin, G. Rowden, and K. A. West (2003)
J. Immunol.
171, 4479-4483
| Abstract »
| Full Text »
| PDF »
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- K. J. Hare, J. Pongracz, E. J. Jenkinson, and G. Anderson (2003)
J. Immunol.
171, 2825-2831
| Abstract »
| Full Text »
| PDF »
- TCR Comodulation of Nonengaged TCR Takes Place by a Protein Kinase C and CD3{gamma} Di-Leucine-Based Motif-Dependent Mechanism.
- C. M. Bonefeld, A. B. Rasmussen, J. P. H. Lauritsen, M. von Essen, N. Odum, P. S. Andersen, and C. Geisler (2003)
J. Immunol.
171, 3003-3009
| Abstract »
| Full Text »
| PDF »
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- I. Tskvitaria-Fuller, A. L. Rozelle, H. L. Yin, and C. Wulfing (2003)
J. Immunol.
171, 2287-2295
| Abstract »
| Full Text »
| PDF »
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- J. B. Chung, A. D. Wells, S. Adler, A. Jacob, L. A. Turka, and J. G. Monroe (2003)
J. Immunol.
171, 1758-1767
| Abstract »
| Full Text »
| PDF »
- LFA-1-induced T cell migration on ICAM-1 involves regulation of MLCK-mediated attachment and ROCK-dependent detachment.
- A. Smith, M. Bracke, B. Leitinger, J. C. Porter, and N. Hogg (2003)
J. Cell Sci.
116, 3123-3133
| Abstract »
| Full Text »
| PDF »
- Linker for Activation of T Cells, {zeta}-Associated Protein-70, and Src Homology 2 Domain-Containing Leukocyte Protein-76 are Required for TCR-Induced Microtubule-Organizing Center Polarization.
- M. R. Kuhne, J. Lin, D. Yablonski, M. N. Mollenauer, L. I. R. Ehrlich, J. Huppa, M. M. Davis, and A. Weiss (2003)
J. Immunol.
171, 860-866
| Abstract »
| Full Text »
| PDF »
- Composition of MHC class II-enriched lipid microdomains is modified during maturation of primary dendritic cells.
- N. Setterblad, C. Roucard, C. Bocaccio, J.-P. Abastado, D. Charron, and N. Mooney (2003)
J. Leukoc. Biol.
74, 40-48
| Abstract »
| Full Text »
| PDF »
- T Cell Glycolipid-Enriched Membrane Domains Are Constitutively Assembled as Membrane Patches That Translocate to Immune Synapses.
- S. Jordan and W. Rodgers (2003)
J. Immunol.
171, 78-87
| Abstract »
| Full Text »
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- Topological Requirements and Signaling Properties of T Cell-activating, Anti-CD28 Antibody Superagonists.
- F. Luhder, Y. Huang, K. M. Dennehy, C. Guntermann, I. Muller, E. Winkler, T. Kerkau, S. Ikemizu, S. J. Davis, T. Hanke, et al. (2003)
J. Exp. Med.
197, 955-966
| Abstract »
| Full Text »
| PDF »
- Low T cell receptor expression and thermal fluctuations contribute to formation of dynamic multifocal synapses in thymocytes.
- S.-J. E. Lee, Y. Hori, and A. K. Chakraborty (2003)
PNAS
100, 4383-4388
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| Full Text »
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- Coxiella burnetii Avoids Macrophage Phagocytosis by Interfering with Spatial Distribution of Complement Receptor 3.
- C. Capo, A. Moynault, Y. Collette, D. Olive, E. J. Brown, D. Raoult, and J.-L. Mege (2003)
J. Immunol.
170, 4217-4225
| Abstract »
| Full Text »
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- The Size of the Synaptic Cleft and Distinct Distributions of Filamentous Actin, Ezrin, CD43, and CD45 at Activating and Inhibitory Human NK Cell Immune Synapses.
- F. E. McCann, B. Vanherberghen, K. Eleme, L. M. Carlin, R. J. Newsam, D. Goulding, and D. M. Davis (2003)
J. Immunol.
170, 2862-2870
| Abstract »
| Full Text »
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- Vav-1 and the IKK{alpha} Subunit of I{kappa}B Kinase Functionally Associate to Induce NF-{kappa}B Activation in Response to CD28 Engagement.
- E. Piccolella, F. Spadaro, C. Ramoni, B. Marinari, A. Costanzo, M. Levrero, L. Thomson, R. T. Abraham, and L. Tuosto (2003)
J. Immunol.
170, 2895-2903
| Abstract »
| Full Text »
| PDF »
- The EphB6 Receptor Inhibits JNK Activation in T Lymphocytes and Modulates T Cell Receptor-mediated Responses.
- A. Freywald, N. Sharfe, C. Rashotte, T. Grunberger, and C. M. Roifman (2003)
J. Biol. Chem.
278, 10150-10156
| Abstract »
| Full Text »
| PDF »
- Involvement of CD70 and CD80 intracytoplasmic domains in the co-stimulatory signal required to provide an antitumor immune response.
- V. Douin-Echinard, J.-M. Peron, V. Lauwers-Cances, G. Favre, and B. Couderc (2003)
Int. Immunol.
15, 359-372
| Abstract »
| Full Text »
| PDF »
- T Cell Receptor Can Be Recruited to a Subset of Plasma Membrane Rafts, Independently of Cell Signaling and Attendantly to Raft Clustering.
- E. Giurisato, D. P. McIntosh, M. Tassi, A. Gamberucci, and A. Benedetti (2003)
J. Biol. Chem.
278, 6771-6778
| Abstract »
| Full Text »
| PDF »
- Naturally Occurring Human IgM Antibody That Binds B7-DC and Potentiates T Cell Stimulation by Dendritic Cells.
- S. Radhakrishnan, L. T. Nguyen, B. Ciric, D. R. Ure, B. Zhou, K. Tamada, H. Dong, S.-Y. Tseng, T. Shin, D. M. Pardoll, et al. (2003)
J. Immunol.
170, 1830-1838
| Abstract »
| Full Text »
| PDF »
- Regulation of Lymphocyte Apoptosis by Interferon Regulatory Factor 4 (IRF-4).
- J. C. Fanzo, C.-M. Hu, S. Y. Jang, and A. B. Pernis (2003)
J. Exp. Med.
197, 303-314
| Abstract »
| Full Text »
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- MHC Class II-Peptide Complexes and APC Lipid Rafts Accumulate at the Immunological Synapse.
- E. M. Hiltbold, N. J. Poloso, and P. A. Roche (2003)
J. Immunol.
170, 1329-1338
| Abstract »
| Full Text »
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- Actin cytoskeletal dynamics in T lymphocyte activation and migration.
- Y. Samstag, S. M. Eibert, M. Klemke, and G. H. Wabnitz (2003)
J. Leukoc. Biol.
73, 30-48
| Abstract »
| Full Text »
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- Live-Cell Dynamics and the Role of Costimulation in Immunological Synapse Formation.
- S. A. Wetzel, T. W. McKeithan, and D. C. Parker (2002)
J. Immunol.
169, 6092-6101
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- Cutting Edge: Association of the Motor Protein Nonmuscle Myosin Heavy Chain-IIA with the C Terminus of the Chemokine Receptor CXCR4 in T Lymphocytes.
- M. Rey, M. Vicente-Manzanares, F. Viedma, M. Yanez-Mo, A. Urzainqui, O. Barreiro, J. Vazquez, and F. Sanchez-Madrid (2002)
J. Immunol.
169, 5410-5414
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- Quantifying signaling-induced reorientation of T cell receptors during immunological synapse formation.
- W. C. Moss, D. J. Irvine, M. M. Davis, and M. F. Krummel (2002)
PNAS
99, 15024-15029
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- Neural and Immunological Synaptic Relations.
- M. L. Dustin and D. R. Colman (2002)
Science
298, 785-789
| Abstract »
| Full Text »
| PDF »
- T cell receptor ligation induces the formation of dynamically regulated signaling assemblies.
- S. C. Bunnell, D. I. Hong, J. R. Kardon, T. Yamazaki, C. J. McGlade, V. A. Barr, and L. E. Samelson (2002)
J. Cell Biol.
158, 1263-1275
| Abstract »
| Full Text »
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- Engagement of the inhibitory receptor CD158a interrupts TCR signaling, preventing dynamic membrane reorganization in CTL/tumor cell interaction.
- N. Guerra, F. Michel, A. Gati, C. Gaudin, Z. Mishal, B. Escudier, O. Acuto, S. Chouaib, and A. Caignard (2002)
Blood
100, 2874-2881
| Abstract »
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- Cutting Edge: Quantitative Imaging of Raft Accumulation in the Immunological Synapse.
- W. R. Burack, K.-H. Lee, A. D. Holdorf, M. L. Dustin, and A. S. Shaw (2002)
J. Immunol.
169, 2837-2841
| Abstract »
| Full Text »
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- Genomic expression programs and the integration of the CD28 costimulatory signal in T cell activation.
- M. Diehn, A. A. Alizadeh, O. J. Rando, C. L. Liu, K. Stankunas, D. Botstein, G. R. Crabtree, and P. O. Brown (2002)
PNAS
99, 11796-11801
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- Essential Role of NF-{kappa}B-Inducing Kinase in T Cell Activation Through the TCR/CD3 Pathway.
- M. Matsumoto, T. Yamada, S. K. Yoshinaga, T. Boone, T. Horan, S. Fujita, Y. Li, and T. Mitani (2002)
J. Immunol.
169, 1151-1158
| Abstract »
| Full Text »
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- TCR Engagement Induces Proline-Rich Tyrosine Kinase-2 (Pyk2) Translocation to the T Cell-APC Interface Independently of Pyk2 Activity and in an Immunoreceptor Tyrosine-Based Activation Motif-Mediated Fashion.
- D. Sancho, M. C. Montoya, A. Monjas, M. Gordon-Alonso, T. Katagiri, D. Gil, R. Tejedor, B. Alarcon, and F. Sanchez-Madrid (2002)
J. Immunol.
169, 292-300
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- Polar Redistribution of the Sialoglycoprotein CD43: Implications for T Cell Function.
- N. D. L. Savage, S. L. Kimzey, S. K. Bromley, K. G. Johnson, M. L. Dustin, and J. M. Green (2002)
J. Immunol.
168, 3740-3746
| Abstract »
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- Intercellular transfer of antigen-presenting cell determinants onto T cells: molecular mechanisms and biological significance.
- D. HUDRISIER and P. BONGRAND (2002)
FASEB J
16, 477-486
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- Cutting Edge: Differential Segregation of the Src Homology 2-Containing Protein Tyrosine Phosphatase-1 Within the Early NK Cell Immune Synapse Distinguishes Noncytolytic from Cytolytic Interactions.
- Y. M. Vyas, H. Maniar, and B. Dupont (2002)
J. Immunol.
168, 3150-3154
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- CD43 polarization in unprimed T cells can be dissociated from raft coalescence by inhibition of HMG CoA reductase.
- E. Gubina, T. Chen, L. Zhang, E. F. Lizzio, and S. Kozlowski (2002)
Blood
99, 2518-2525
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- Focal Localization of Placental Protein 14 Toward Sites of TCR Engagement.
- J. Rachmilewitz, Z. Borovsky, G. Mishan-Eisenberg, E. Yaniv, G. J. Riely, and M. L. Tykocinski (2002)
J. Immunol.
168, 2745-2750
| Abstract »
| Full Text »
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- A Novel Serine-rich Motif in the Intercellular Adhesion Molecule 3 Is Critical for Its Ezrin/Radixin/Moesin-directed Subcellular Targeting.
- J. M. Serrador, M. Vicente-Manzanares, J. Calvo, O. Barreiro, M. C. Montoya, R. Schwartz-Albiez, H. Furthmayr, F. Lozano, and F. Sanchez-Madrid (2002)
J. Biol. Chem.
277, 10400-10409
| Abstract »
| Full Text »
| PDF »
- The role of lipid rafts in signalling and membrane trafficking in T lymphocytes.
- M. A. Alonso and J. Millan (2002)
J. Cell Sci.
114, 3957-3965
| Abstract »
| Full Text »
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- How and Why Does the Immunological Synapse Form? Physical Chemistry Meets Cell Biology.
- A. K. Chakraborty (2002)
Sci. STKE
2002, pe10
| Abstract »
| Full Text »
| PDF »
- Rho GTPases link cytoskeletal rearrangements and activation processes induced via the tetraspanin CD82 in T lymphocytes.
- A. Delaguillaumie, C. Lagaudriere-Gesbert, M. R. Popoff, and H. Conjeaud (2002)
J. Cell Sci.
115, 433-443
| Abstract »
| Full Text »
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- Cutting Edge: Negative Regulation of Immune Synapse Formation by Anchoring Lipid Raft to Cytoskeleton Through Cbp-EBP50-ERM Assembly.
- K. Itoh, M. Sakakibara, S. Yamasaki, A. Takeuchi, H. Arase, M. Miyazaki, N. Nakajima, M. Okada, and T. Saito (2002)
J. Immunol.
168, 541-544
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