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V
3-integrin
Departments of 1 Physiology and
2 Rheumatology, During inflammation neutrophils are recruited
from the blood onto the surface of microvascular endothelial cells. In
this milieu the presence of soluble chemotactic gradients is disallowed by blood flow. However, directional cues are still required for neutrophils to migrate to the junctions of endothelial cells where extravasation occurs. Shear forces generated by flowing blood provide a
potential alternative guide. In our flow-based adhesion assay
neutrophils preferentially migrated in the direction of flow when
activated after attachment to platelet monolayers. Neutralizing
integrin signaling; adhesion molecule cross talk; regulated
migration
NEUTROPHILS ARE RECRUITED from flowing blood and
localized at inflammatory sites by an orchestrated sequence of adhesive
and activation steps that occur on the surface of microvascular
endothelial cells (see Refs. 17, 30, 35 for review). Briefly,
neutrophils are captured from flow by members of the selectin family of
adhesion molecules that support the rolling adhesion of tethered cells. Rolling adhesion slows the neutrophils down and allows the transmission of chemotactic triggering signals from the endothelium. Neutrophil Activation of neutrophils by chemotactic agents drives the process of
neutrophil migration by inducing actin filament assembly in the
cytoskeleton and promoting the rapid turnover of
Recently, we studied neutrophil adhesion to activated immobilized
platelets in a flow-based assay and reconstructed adhesive phenomena on
immobilized purified adhesion receptors (27, 28). Adhesion to platelet
monolayers may represent the situation during thrombotic pathology, but
it is also relevant to neutrophil migration on endothelium because
adhesion molecules such as P-selectin, CD31 [platelet endothelial
cell adhesion molecule (PECAM)-1], and intercellular adhesion
molecule (ICAM)-2 are expressed on both platelets and endothelium. We
demonstrated that ligation of surface-presented P-selectin or CD31
caused neutrophils to increase the velocity of their
CD11b/CD18-mediated migration (27). While analyzing migration
velocities we noticed that neutrophils tended to move preferentially in
a direction parallel to the fluid flowing over them. We hypothesized
that fluid shear forces were directing migratory effort and that
specific interactions between a surface-presented molecule and a
neutrophil receptor transduced information on the direction of shear
force into the cell. On investigation, the transducer of flow-directed
migration was found to be neutrophil
Neutrophil Isolation
ABSTRACT
Top
Abstract
Introduction
Methods
Results
Discussion
References
V
3-integrin
with monoclonal antibodies or turning the flow off randomized the
direction of migration without affecting migration velocity. Purified,
immobilized
V3-integrin
ligands, CD31 and fibronectin, could both support flow-directed
neutrophil migration in a concentration-dependent manner. Migration
could be randomized by neutralizing
V3-integrin
interactions with the substrate using antibodies or
Arg-Gly-Asp-containing peptide. These results exemplify
mechanical signal transduction through integrin-ligand interactions and
reveal a guidance system that was hitherto unknown in neutrophils. In
more general terms, it demonstrates that cells can use integrin
molecules to "sample" their physical microenvironment through
adhesion and use this information to modulate their behavior.
INTRODUCTION
Top
Abstract
Introduction
Methods
Results
Discussion
References
2-integrins, activated as a
consequence of the triggering signals, then mediate stationary adhesion
by binding to members of the immunoglobulin superfamily (IgSF)
expressed on the endothelial surface. The process culminates with the
integrin-mediated migration of neutrophils into the extravascular tissue.
2-integrin adhesion molecules
that support locomotion (15, 16, 28, 31). These dynamic changes can be
regulated in response to cues such as gradients of soluble chemotactic
agents in the environment (24, 34). Gradients are "sensed" by
specific membrane-spanning receptors that allow cells to orientate and
undertake directed migration. Clearly, neutrophils activated in the
vessel lumen require guidance to the margins of endothelial cells where
the migration to extravascular tissues occurs. However, concentration gradients of soluble activating agents would be disrupted by blood flow
and therefore cannot be utilized in the lumen of blood vessels.
V3-integrin
interacting with surface-anchored ligand(s). Not only does this
represent a novel means of neutrophil guidance, but it is also an
example of a more general mechanism by which cells "sample"
their physical environment and modulate their behavior accordingly.
METHODS
Top
Abstract
Introduction
Methods
Results
Discussion
References
Immobilization of Platelet Monolayers in Microslides
Microslides (Cam Lab, Cambridge, UK) are glass capillary tubes with a rectangular cross section of 0.3 × 3 mm, a length of 5 cm, and good optical qualities. To provide a substrate that readily binds platelets and purified adhesion receptors, microslides were coated with 3-aminopropyltriethoxysilane (APES; Sigma) as previously described (7, 27). Blood for platelet preparation was collected into heparin sodium (CP Pharmaceuticals, Wrexham, UK). Platelet-rich plasma was derived by spinning the blood at 400 g for 5 min and removing the platelet-rich supernatant. Platelets were counted, adjusted to 2 × 108/ml with PBS, loaded into microslides, and incubated at room temperature for 40 min to allow them to sediment, adhere, and form a confluent monolayer (27, 28). Unbound platelets were removed by washing before assay. Experiments were always conducted using autologous platelets and neutrophils.Immobilization of Purified Adhesion Receptors in Microslides
Affinity-purified recombinant CD31, P-selectin, ICAM-1 (all produced from stably transfected Chinese hamster ovary cells; gift of Ian Collins, R and D Systems, Abingdon, UK) and fibronectin (derived from human fibroblasts; Calbiochem), were dissolved in PBS. The proteins were immobilized by incubating the desired concentration in an APES-treated microslide for 60 min at 37°C. The microslides were washed, and unoccupied protein-binding sites were blocked with 1% BSA in PBS. The surface density of purified CD31 and P-selectin immobilized in microslides has been previously quantified and ~3 µg/ml found to give a level similar to expression on platelet monolayers (27).Adhesion and Migration Under Conditions of Flow
The adhesion assay was based on that recently described (7, 27, 28). Microslides containing confluent platelet monolayers or purified adhesion receptors were attached to a Harvard syringe pump by means of flexible silicon tubing, and cell-free buffer or a suspension of neutrophils was drawn through the microslides via a microelectronic switching valve (Lee Products, Gerards Cross, UK). A constant wall shear stress (0.05 Pa) was maintained in the microslide by choice of the appropriate flow rate. This equates to 0.5 dyn/cm2 and is at the low end of the physiological range of wall shear stress in postcapillary venules (21).Microslides were mounted on the stage of a microscope fitted with a
video camera, monitor, and recorder. Neutrophil suspension was flowed
over the adhesive substrate for 5 min. Nonadherent cells were washed
from the microslide by perfusing cell-free buffer (0.1% BSA in PBS)
for 2 min. Rolling neutrophils were then activated on the adhesive
surface by the continuous perfusion of
N-formylmethionyl-leucyl-phenylalanine (FMLP)
(10
7 M). On exposure to
chemotactic stimuli, neutrophils stopped rolling, underwent shape
change, and spread on the platelet surface (27, 28). After the initial
period of spreading, neutrophils began to migrate across the platelet
surface (27).
The direction and velocity of migration was analyzed. A single field of
rolling neutrophils was chosen and the response to chemotactic
stimulation recorded by time-lapse videomicroscopy (using a ninefold
time condensation) for a duration of 10 min. In initial experiments
conducted on immobilized platelets, the rate of migration was assessed
offline for 20 cells in each microscope field; however, in later
experiments we found that analysis of 10 migrating cells provided
results that were not quantitatively different from data from 20 cells.
Analysis of migration velocities (in µm/min) has been described in
detail previously (27). The distance migrated in the direction of flow
over the 10-min period was measured for each cell (Fig.
1; note that both positive and negative
values are possible depending on whether individual cells moved with or
against flow). Because the migration velocity of neutrophils has
previously been demonstrated to vary with the composition of the
adhesive substrate (27), the distance moved in the direction of flow
was normalized for migration velocity. Thus the distance moved in the
direction of flow was expressed as a percentage of the total distance
migrated during the 10 min for each cell to give a directional index of
migration.
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When P-selectin was not present on the microslide (i.e., when comparing
directed migration on purified CD31 without purified P-selectin), there
was no rolling adhesion, so it was necessary to activate neutrophils
with FMLP (107 M)
immediately before perfusion. To allow the direct adhesion of flowing
neutrophils via activated integrins, low wall shear stress (0.0125 Pa)
was applied for 1.5 min of cell perfusion. The wall shear stress was
then returned to 0.05 Pa, and a continuous video record of migration
was made from exactly 2 min after initial activation. In this way we
were able to follow migration from 2 to 10 min after activation. The
velocity and the index of directional migration were analyzed as before.
Antibody and Peptide Interventions
Neutrophils were treated with monoclonal antibodies (MAbs), Arg-Gly-Asp (RGD) peptide, or control peptide for 15 min before perfusion. Peptides were also perfused with the buffer containing the FMLP. All monoclonal antibodies were isotype IgG1 unless otherwise stated. MAbs against CD31 were function-blocking, L133.1-recognizing domain 2 (Beckton Dickinson, Oxford, UK) and 7E4-recognizing domain 3 (designated A082, from the 6th International CD Antigen Workshop; isotype IgG2), and nonblocking controls 10B8-recognizing domain 5 (gift of Ian Collins, R and D Systems) and 9G11-recognizing domain 1 (10); all were used at 10 µg/ml. MAb againstV-integrin was L230
(from ATCC), which blocked interaction with CD31 and fibronectin (10)
and was used at 18 µg/ml. Antibody against 3-integrin was clone RUU-PL
7F12 (Becton Dickinson) used at 10 µg/ml. Antibody against
V3-integrin
was 6C2 used at 10 µg/ml (4). As a control we used an isotype-matched
antibody against CD11a (MAb DA36 was a gift of Martyn Robinson
Celltech, Slough, UK), another neutrophil-borne integrin molecule. RGD
and control peptide were Arg-Gly-Asp-Ser and Arg-Gly-Glu-Ser,
respectively (Sigma), and were used at 500 µM. To ensure that
treatment of neutrophils with antibody or RGD peptide did not itself
activate or modify activation of neutrophils, the expression of the
neutrophil integrin CD11b (which supports migration) was measured by
fluorescence-activated cell sorting (FACS 440, Becton Dickinson). Cells
were either unstimulated or activated by FMLP
(107 M) in the presence of
the anti-V-integrin antibody
L230 or RGD peptide and labeled with
R-phycoerythrin-conjugated anti-CD11b MAb (Dako,
High Wycombe, UK). In two experiments, neither reagent significantly
altered the levels of expression of CD11b in resting or FMLP-stimulated
cells (values within 5% of controls without L230 or RGD).
Statistics
Effects of different treatments were tested using analysis of covariance, and individual treatments were tested by one-way analysis of variance. Comparison of individual treatments were made by paired t-test or unpaired (Student's) t-test as appropriate.| |
RESULTS |
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Top
Abstract
Introduction
Methods
Results
Discussion
References
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Flow-Directed Neutrophil Migration on Platelets
Using a flow-based adhesion assay and neutrophils adherent to platelet monolayers, we investigated the direction of neutrophil migration. When neutrophils were perfused over platelet monolayers for 5 min, ~1,500/mm2 became adherent. Of these, >90% were rolling with a mean velocity of 1.9 ± 0.15 µm/s (mean ± SE of 60 cells from 3 experiments). When they were activated with FMLP all the neutrophils rapidly came to halt, changed shape, spread, and migrated over the surface of the platelets (27, 28). Neutrophils, however, were not migrating at random. Rather, the direction of their locomotion was influenced by the shear forces generated by flow (Fig. 1). For example, detailed analysis showed that the 10 neutrophils in Fig. 1 had a mean migration velocity (averaged over 10 min at 1-min intervals) of 10.0 ± 1.1 µm/min and an average movement downstream of 4.0 ± 1.3 µm/min. Thus the cells had a mean directional index of migration of 40%. The response of individual cells varied widely. For instance, neutrophil 3 (Fig. 1) had a directional index of 73% (downstream movement = 9.5 µm/min, velocity = 13.1 µm/min), whereas neutrophil 6 had a directional index of migration of
17% (downstream movement = 1.6 µm/min, velocity = 9.2 µm/min) and demonstrated a net movement in the opposite direction
to flow. On average over 10 experiments, samples of neutrophils
migrating on platelets in the presence of flow showed a directional
index of migration between 30 and 50%, with an average of 85 ± 12%
of migrating cells showing net displacement in the direction of flow.
We demonstrated that directional migration was due to flow by turning
off the perfusion of buffer. This randomized the direction of migration
so that 41 ± 7% of neutrophils moved in what was previously the
direction of flow. Turning the flow off did not affect the velocity of
neutrophil migration. Thus, in a series of three comparative
experiments, neutrophils migrated with a mean velocity of 9.6 ± 0.2 µm/min in flow and 9.8 ± 0.5 µm/min in the absence of
flow, while the directional index of migration was 48 ± 3% and
3 ± 1%, respectively (all data are means ± SE of means from
3 experiments). In two experiments we turned flow back on after cells
had migrated in random directions for 10 min, and flow-directed
migration was reestablished (data not shown). We also investigated the
effect of increasing wall shear stress from 0.05 to 0.2 Pa and found
that the directional index of migration was unaffected, averaging 25%
at 0.05 Pa and 29% at 0.2 Pa (means from 3 experiments).
Role of
V-Integrin in
Directing Neutrophil Migration on Platelets
V-integrin, migration
was no longer directional (Fig. 2).
Blockade of another neutrophil integrin CD11a
(L-integrin) had no effect on
direction. Thus an integrin containing the
V-subunit was essential for
flow-directed neutrophil migration. More specifically, V3-integrin
was the probable heterodimer utilized, because the reported alternative
associations of V- with the
1-,
5-, and 6-subunits (see Ref. 25 for
review) have not been demonstrated on neutrophils whereas
V3-integrin
has (11). Activation of platelets brings fibrinogen, fibronectin,
thrombospondin, and von Willebrand factor to the platelet surface (12),
all of which are potential ligands for
V3-integrin
(25). CD31 (PECAM-1), an IgSF adhesion molecule presented on resting
and stimulated platelets, is also a ligand for
V3-integrin
(5, 26). The multiplicity of platelet ligands for
V3-integrin
made functional blockade studies on platelets problematic. Thus, for
instance, in the presence of a panel of function-blocking (A082 and
L133.1) and control (9G11 and 10B8) MAbs against CD31, there was no
consistent effect on directional migration (Fig.
3). Either CD31 was not a ligand for
V3-integrin
in this system or other ligands could be efficiently utilized
simultaneously.
V3-Integrin
binds fibrinogen and fibronectin by recognition of RGD amino acid
motifs in the protein structure (14, 25). However, when we perfused the adhesion-blocking peptide containing an RGD motif along with
neutrophils, the platelet monolayer was destabilized, leading to the
loss of the adhesive substrate.
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Directed Neutrophil Migration on Purified Adhesion Receptors
Migration on purified P-selectin. Because monolayers of activated platelets were too complex a substrate for investigating the adhesive requirements for flow-directed migration, we used combinations of purified adhesion receptors immobilized in microslides. When we immobilized 1 µg/ml of purified recombinant P-selectin in the microslide and saturated all additional protein binding sites with albumin, neutrophils were captured from flow and on activation migrated over the surface of the immobilized protein utilizing albumin as a ligand (13, 27) with a velocity of 7.8 ± 0.3 µm/min. Importantly, there was no directional component to migration (directional index of migration = 3 ± 5%; mean ± SE of means from 3 experiments).Migration on CD31 and P-selectin. In
matched experiments in which 5 µg/ml of CD31 was coimmobilized with
the 1 µg/ml P-selectin, neutrophils migrated with a velocity of 11.8 ± 0.4 µm/min and directional index of migration of 38 ± 2%. Thus
CD31 not only increased the velocity of neutrophil migration (27) but
also enabled transduction of a directional cue. In the presence of antibody against V-integrin,
neutrophils could not utilize purified CD31 to orientate migration
(directional index of migration = 3 ± 4%; mean ± SE of means from
3 experiments).
Migration on CD31 alone. In a separate
series of experiments the amount of CD31 immobilized in microslides was
varied between 1 and 5 µg/ml, and it was found that the directional
index of migration increased in parallel with CD31 concentration (Fig. 4). The ability of the neutrophil to
utilize CD31 for directional migration in combination with the ability
of anti-V MAb to ablate the
phenomenon strongly supports the conclusion that
V3-integrin is the sensing receptor because other integrin heterodimers containing the V-subunit do not utilize
CD31 as a counter ligand (25).
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Migration on fibronectin and
P-selectin. We also demonstrated the ability of an
alternative
V3-integrin
ligand, fibronectin, to direct migration. Figure
5 shows that the directional
index of migration of neutrophils was dependent on the concentration of
fibronectin immobilized with 1 µg/ml P-selectin. The velocity of
neutrophil migration on 2.5 µg/ml of fibronectin was 8.2 ± 1.0 µm/min and did not vary significantly with fibronectin
concentration (mean ± SE of means from 3 experiments). In the
presence of MAb against
V-integrin, directional
migration was abolished (directional index of migration = 3 ± 5% compared with 38 ± 2% for untreated cells; mean ± SE
of 3 experiments using 2.5 µg/ml fibronectin). To verify that
V3-integrin
was the heterodimer used to sense shear stress, we used antibodies
against 3-integrin and
V3-integrin. In two experiments with each antibody and using 2.5 µg/ml
fibronectin, the directional index of migration was reduced from 33%
for control cells to 1% or 6% for cells treated with
anti-3 or
anti-V3, respectively. Additionally, the perfusion of neutrophils with RGD
peptide completely abolished directional migration on fibronectin, whereas a control peptide had no effect (Fig.
6). We also conducted experiments using
ICAM-1 as a ligand for other integrin heterodimers. In two experiments
neutrophils did not show flow-directed migration on ICAM-1 immobilized
with albumin (data not shown). Importantly, in all of the experiments
on purified adhesion receptors, RGD peptide or MAb against
V-integrin had no effect on the
level of neutrophil adhesion to the substrate, neutrophil spreading, or
migration velocity (data not shown), processes that have been demonstrated to be dependent on
2-integrin (24, 27, 28, 34).
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DISCUSSION |
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Top
Abstract
Introduction
Methods
Results
Discussion
References
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In the current study we demonstrate a novel route by which the
V3-integrin
of neutrophils can influence the direction of neutrophil migration,
which is itself supported by
M2-integrin (CD11b/CD18; MAC-1) (9, 27, 28). The direction of flow was
"sensed" by neutrophil
V3-integrin
when it was bound to substrate, presumably by the transmission of
forces exerted on the integrin-ligand bond. Monolayers of activated,
immobilized platelets were able to support directed neutrophil
migration but presented too many potential ligands to allow conclusive
demonstration of the counter-receptor for
V3-integrin.
Using purified, immobilized adhesion receptors, it was apparent that
V3-integrin
could utilize surface-bound CD31 or fibronectin to direct migration.
Furthermore, the degree of alignment between migration and flow was
dependent on the concentration of CD31 or fibronectin immobilized, but
not critically on the magnitude of the shear forces applied to adherent cells, although alignment was rapidly lost when flow was stopped. Thus
a mechanical signal transduced through
V3-integrin
was capable of directing cytoskeletal rearrangement and the adhesive interactions of 2-integrin(s)
that underlie neutrophil migration in this model (9, 27, 28). These
results exemplify the use of integrin binding as a guidance system that
was hitherto unknown in neutrophils.
Separate Roles for
V3-Integrin
and
M2-Integrin
in Migrating Neutrophils
V3-integrin
was not directly supporting the adhesion or the migration of
neutrophils. Thus adhesion-blocking MAb against
V-integrin or adhesion-blocking RGD peptide had no effect on the number of neutrophils bound to or
actively migrating over adhesive substrates, nor did such treatment influence the velocity of migration (Ref. 27 and current study). Conversely, we previously demonstrated that in the presence of MAb
against M-integrin (CD11b) or
2-integrin (CD18), migration was abolished and >90% of neutrophils detached from a surface of
immobilized activated platelets after activation under conditions of
flow (27, 28). Evidently,
V3-integrin
was unable to resist the shear forces generated by flow in the absence
of binding through M2-integrin
adhesion and thus did not function as a primary anchorage molecule.
Interestingly, chelation of intracellular calcium has been shown to
increase
V3-integrin
affinity for vitronectin and disallow onward migration of neutrophils
(11). Thus
V3-integrin
appears capable of mediating strong adhesion to the substrate, but in
the current model "weak" interactions between
V3-integrin
and the substrate fulfilled a signaling rather than adhesive role
during neutrophil migration.
Ligation of
V3-integrin
was essential for efficient migration of avian embryonic neural crest
cells (8), human and rat smooth muscle cells (2, 20, 23), and human
melanoma cells (1). Inhibition of interactions between
V3-integrin
and its ligand(s) ablated migration but not adhesion to the substrate. Thus
V3-integrin
regulated migration in these cells by a process of "cross talk"
with other integrin heterodimers (e.g.,
V1
and V5)
that supported primary adhesion and migration. Interestingly, V3-integrin
appears to retain the capacity to enhance the function of other
integrin molecules when transfected into null cells. Thus signaling
through
V3-integrin
inserted into the monocytic cell line K562 was essential for the
phagocytosis of fibronectin-coated beads via endogenously
expressed
51-integrin
(3). Similarly, signaling via transfected
V3-integrin
was essential for
51-integrin-mediated migration toward a source of fibronectin in human embryonic kidney cell
line HEK-293 (29).
With regard to leukocyte migration, Imhof and Dunon (17) recently
demonstrated that occupancy of lymphocyte
V3-integrin by either CD31 or vitronectin increased the rate of cell motility supported by
41-integrin
(VLA4) using vascular cell adhesion molecule-1 as a counter-ligand
(18). This presents an interesting contrast with neutrophils, which did
not use
V3-integrin
as an "accelerator" when activated on platelet
monolayers (27). Rather, neutrophils regulated the velocity of their
migration using accessory signals from the homophilic interaction of
neutrophil CD31 with surface-presented CD31 and/or neutrophil
P-selectin glycoprotein ligand-1 with surface-presented P-selectin
(27). Thus signals from surface-presented "adhesion" molecules
have the potential to maximize the efficiency of neutrophil migration into inflamed tissues. In combination with our previous demonstration that surface-presented CD31 and P-selectin could modulate the migration
velocity of neutrophils, we have demonstrated the ability of these
cells to simultaneously integrate directional signals from
surface-presented ligands of
V3-integrin.
The ability of such signals to modify cellular responses illustrates
how the physical microenvironment may play a critical role in
regulating cellular physiology. Furthermore, and considering the above
examples, outside-in signaling through
V3-integrin
may be a generalized route by which the motility of cells is regulated.
The exact nature of cellular responses and the identity of the
receptors to which V3-integrin
"talks" appear to be strongly dependent on cell type and the
composition of the adhesive substrate.
V3-Integrin
as a Physiological Sensor of the Physical Environment
V3-integrin
modulated migration by acting as a sensor of mechanical stress. Another
study has shown that cyclical mechanical forces caused mitogenic
changes in smooth muscle cells from rat aorta that were abrogated by
antibodies recognizing the 3-integrin subunit or the
V5-integrin
heterodimer (33). Thus interactions of these integrins with matrix
elements appeared essential for the transduction of mechanical force
into the cells. The concept of the "hard-wired cell" in which a
structural scaffolding linked to surface integrins transmits
information about the physical environment into the cell interior has
received recent attention (see Ref. 19 for review). Direct application
of force to surface integrins has been shown to induce remodeling of
the cytoskeleton and relocalization of intracellular elements such as
the apparatus for protein synthesis (6, 32). Mechanical forces acting
through focal adhesion complexes are thought to be able to modify the response of signaling elements assembled there (19). The present study
is a new example of a highly dynamic physiological response that
conforms to this concept, with rapid and reversible modification of
neutrophil behavior driven by stress applied to an integrin-substrate bond.
Here, the mechanically transduced signal was integrated with that from
the FMLP receptor, presumably to direct cyclic changes in the actin
cytoskeleton and binding of the
2-integrins, which form the
primary migratory apparatus. The signaling routes by which this
integration is achieved are not clear, but modulation of intracellular
calcium levels is important in the function of V3-integrin
(11). On a physical level, a polarized location of
V3-integrin
at the leading edge of neutrophils has been demonstrated for
neutrophils migrating on vitronectin but not on fibronectin (22). Thus,
although it is attractive to suggest that a localized signaling complex
would be well placed to regulate direction of migration (e.g., by
preferentially acting as a focus for actin polymerization), evidence
for such a mechanism in neutrophils is lacking. Regardless of the
actual signal induced by the transduction of a force through
V3-integrin,
and indeed the means by which this modifies migration, the
physiological implication is that neutrophils can align their migration
in parallel with the direction of circulating blood and avoid wasted
effort in random movement before passing between endothelial cells.
Recently, we also demonstrated that the velocity of neutrophil
migration is increased by the presence of CD31 and P-selectin on the
adhesive substrate (27). Thus we suggest that accessory signals derived
from adhesive interaction with the substrate maximize the efficiency of
neutrophil migration by accelerating and directing locomotion.
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FOOTNOTES |
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The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Address reprint requests to G. E. Rainger.
Received 17 July 1998; accepted in final form 3 November 1998.
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REFERENCES |
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Top
Abstract
Introduction
Methods
Results
Discussion
References
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30% (Control). In the presence of monoclonal antibody (MAb)
against 
