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Blood, Vol. 92 No. 8 (October 15), 1998:
pp. 2951-2958
Sickle Cell Adhesion to Laminin: Potential Role for the  5 Chain
By
Sheritha P. Lee,
Michelle L. Cunningham,
Patrick C. Hines,
Christopher C. Joneckis,
Eugene P. Orringer, and
Leslie V. Parise
From the Department of Biology, North Carolina Central University,
Durham, NC; and the Departments of Pharmacology and Medicine, and The
Center for Thrombosis and Hemostasis, The University of North Carolina
at Chapel Hill, Chapel Hill, NC.
 |
ABSTRACT |
Sickle red blood cell (RBC) adhesion to the endothelium and to
exposed, underlying subendothelial proteins is believed to contribute
to vascular occlusion in sickle cell disease. Laminin, a major
component of the subendothelium, supports significant adhesion of
sickle, but not normal RBCs. The purpose of this study was to define
the adhesive region for sickle RBCs within a human laminin preparation
using a flow adhesion assay designed to mimic physiologic flow through
postcapillary venules. Because sickle RBCs did not adhere to the common
laminin contaminants entactin or collagen type IV, neither of these
proteins are likely to contribute to the observed adhesion to laminin.
Known adhesive regions of laminin neither supported nor inhibited
sickle RBC adhesion to laminin, suggesting a mechanism of adhesion
previously uncharacterized in other laminin adhesion studies. Moreover,
sickle RBCs did not adhere to mouse EHS laminin or to human laminin-2
(merosin), eliminating the  1, 2,  1, and  1 chains as
mediators of sickle cell adhesion. The monoclonal antibody 4C7, which
binds at or near the G-domain of the laminin 5 chain, significantly
inhibited sickle RBC adhesion. These results suggest that an adhesive
region for sickle RBCs is contained within the laminin 5 chain.
© 1998 by The American Society of Hematology.
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INTRODUCTION |
THE HALLMARKS OF sickle cell disease are
vascular occlusion and painful crises.1 Although crises
vary among patients in severity, frequency, and duration, they are all
periodic, acute episodes of pain believed to be caused by local
disruptions of blood flow with possible tissue ischemia or
infarction.1 The homozygous state for the sickle mutation
(SS) generally has the most severe clinical course,2 which
may ultimately lead to organ failure and death.3
The disruption of blood flow that leads to vascular occlusion may occur
in large vessels4,5 but is most commonly associated with
the microvasculature. Whereas erythrostasis due to the rigidity of
polymerized S hemoglobin (Hb S) contributes to
vaso-occlusion,6 erythrostasis alone is probably
insufficient to explain the extent of vascular occlusion seen in
patients.7,8 Sickle red blood cells (RBCs) have been shown
to adhere to the endothelium in vitro9,10 and may
contribute to vessel occlusion in vivo by delaying RBC travel time,
leading to greater decreases in oxygen tension, increased cell
sickling,11,12 and endothelial damage.
In addition to endothelial cell adhesion, sickle cells may also adhere
to subendothelial proteins, because factors such as increased levels of
tumor necrosis factor (TNF), fibrin/fibrinogen degradation
products, and thrombin generation exist in sickle cell disease and are
known to retract endothelial cells, causing subendothelial protein
exposure in vitro.13-18 In fact, sickle cell patients have
greater levels of circulating endothelial cells19-21 and
extensive vascular damage upon postmortem examination.22
Once endothelial integrity has been compromised, extracellular matrix
(ECM) components such as laminin, thrombospondin, fibronectin, von
Willebrand factor, vitronectin, and collagen are potentially exposed to
sickle RBCs. Abnormal adhesion between sickle RBCs and thrombospondin
under static and flow conditions has been documented,23,24 as well as even greater levels of adhesion to the ECM protein laminin.24 However, the mechanism of adhesion or the region of laminin that is adhesive to sickle cells is unknown.
Laminin is a multidomain, heterotrimeric protein composed of one each
of the five known , three known , and two known chains.25 The chains complex in various combinations to
form a characteristic cross-shape of three short arms and one long arm
with a large globular domain at its C-terminus.26 Human placental laminin preparations likely contain a mixture of laminin isoforms, in addition to common contaminants entactin and collagen type
IV.27
In this study we sought to better characterize the adhesion of sickle
cells to laminin, to identify the component of the laminin preparation
that is adhesive for sickle cells, and to identify the adhesive region
within this component. We not only eliminate many likely adhesive
sites, but also provide evidence that the laminin 5 chain, at or
near the G-domain, contains an adhesive region for sickle cells, based
on the inhibition studies with the 4C7 monoclonal antibody (MoAb).
Because the laminin 5 chain is present in the
subendothelium,25 this chain may contribute to
vaso-occlusion suffered by sickle cell patients.
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MATERIALS AND METHODS |
Reagents.
Human placental laminin and human collagen type IV were obtained from
GIBCO (Grand Island, NY). Recombinant human entactin was generously
provided by R. Timpl (Max-Planck Institute, Martinsried, Germany). The antihuman laminin polyclonal antibody (pAb)
was purchased from Upstate Biotechnology Inc (Lake Placid, NY). The MoAb 4C7 was purchased from GIBCO and Chemicon (Temecula, CA). Protein
G Sepharose beads were obtained from Pharmacia Biotech (Piscataway,
NJ). Synthetic laminin peptides of known adhesive regions of laminin
were kindly provided by H. Kleinman (National Institutes of Health,
Bethesda, MD). Hank's Balanced Salts Solution (HBSS) was obtained from
Sigma Chemical Co (St Louis, MO).
RBC preparation.
Sickle RBCs were obtained from homozygous sickle cell patients (Hb SS)
during clinic visits to the UNC Comprehensive Sickle Cell Center.
Normal RBCs (Hb AA) were obtained from healthy volunteers. All blood
samples were drawn by venipuncture into 0.13 mol/L sodium citrate and
processed immediately upon venipuncture by centrifuging at 150g
for 15 minutes to isolate blood cells from plasma and platelets. RBCs
were then washed three times in CGS (1.29 mmol/L sodium citrate, 3.33 mmol/L glucose, 124 mmol/L NaCl, pH 7.2), then resuspended in
phosphate-buffered saline (PBS; 137 mmol/L NaCl, 10.2 mmol/L
NaH2PO4, 1.76 mmol/L
KH2PO4, 2.68 mmol/L KCl), and packed by
centrifugation at 800g for 10 minutes. A 1% hematocrit was
then prepared by diluting 30 µL of packed RBCs per 1.5 mL of
perfusion media (HBSS supplemented with 0.3% bovine serum albumin [BSA] and 20 mmol/L HEPES, pH 7.4).
For experiments shown in Table 1, RBC
density fractions were prepared from unpacked RBCs using a modified
arabinogalactan density gradient, using a high potassium isotonic
buffer as previously modified23 from Sorette et
al,28 an approach designed to minimize reticulocyte
dehydration. RBCs from the top 20% and bottom 5% of the gradient were
used. We found by methylene blue staining of RNA that reticulocytes
made up 20% to 90% of the top fraction and less than 5% of the
bottom fraction. Nucleated RBCs were not observed in Wrights stain of
the isolated fractions. By trypan-blue exclusion, approximately 90% of
the isolated RBCs were intact. Experiments were performed immediately
after RBC isolation.
Antibody purification.
Protein G Sepharose beads were washed and resuspended in an equal
volume of PBS to create a 50% slurry. This slurry (50 mL) was then
added to the 4C7-ascites solution and inverted slowly at 4°C for 18 hours. After collecting the cleared ascites fluid, the beads were
washed twice in PBS and then treated with 0.1 mol/L glycine  HCl,
pH 2.7, to elute the 4C7 MoAb. Tris, pH 9.0, was added to neutralize
the purified antibody, which was stored at 4°C until use.
Flow adhesion assay.
The flow adhesion system used for this study was designed by J. Moake
and L. McIntire (Rice University, Houston, TX) to mimic blood flow
through postcapillary venules, as described.23 The protein
of interest (0.75 µg) was immobilized onto each of the two identical
wells in the silicon gasket by incubating at 4°C overnight, thus
allowing two separate conditions to be performed during each
experiment.
A 1% hematocrit solution (1.5 mL) was made to flow over wells at a
rate of 1.0 mL/min and constant shear stress of 1 dyne/cm2.
This shear stress has been measured in postcapillary
venules,29 in which some investigators think vaso-occlusion
occurs29,30 and in which the greatest in vitro sickle cell
adhesion has been measured.31-33 After a 6-minute wash
period, the number of adhered cells from 4 representative areas of the
well were counted from a 0.25-mm2 field, normalized,
averaged, and presented as cells per square millimeter ± SD. Any
white blood cells that may have adhered were not counted.
Inhibition assays.
Potential inhibitors of adhesive regions of laminin or the appropriate
control were added to the wells containing the immobilized protein and
incubated at 37°C for 2 hours. Wells were then washed with PBS
before continuing the flow adhesion assay. Potential inhibitors of
laminin receptors on RBCs were tested by adding the inhibitor to the
1% hematocrit solution and incubating at 37°C for at least 30 minutes before the flow adhesion assay.
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RESULTS |
Using a microvessel flow adhesion system, we measured sickle RBC
adhesion to the ECM protein laminin isolated from human placenta (hLAM). Laminin supported approximately 3,000 adherent sickle cells/mm2, but did not support the adhesion of normal RBCs
(Fig 1A). These results suggest that
laminin contains an adhesive site for RBCs that may be unique to the
sickle cell disease state.
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| Fig 1.
Sickle RBCs adhere to laminin. A 1% hematocrit of sickle
(SS) RBCs or normal (AA) RBCs was flowed across immobilized protein
(0.75 µg). Adherent cells from four representative regions were
counted by microscopy, normalized to cells per square millimeter, and
shown as the mean ± SD. (A) Significant adhesion of sickle, but not
normal, RBCs was observed to human laminin (hLAM). Only background
levels of adhesion were observed with sickle RBCs to the negative
control BSA. Data from 10 sickle cell patients are shown. (B) Sickle
RBCs were flowed across immobilized recombinant human entactin (ENT) or
collagen IV (Coll IV). SS RBCs did not adhere to either entactin or
collagen IV. BSA and hLAM are shown as negative and positive controls,
respectively. Data from 3 patients are shown. (C) Immobilized hLAM was
pretreated with an antilaminin polyclonal antibody (pAb) before the
introduction of the SS 1% hematocrit. The antilaminin pAb
significantly inhibited the adhesion of sickle RBCs to hLAM. Adhesion
to laminin in the absence of an antibody (hLAM) and in the presence of
a nonspecific, isotype-matched IgG antibody are shown as negative
controls. Data shown represent experiments from 4 patients.
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Human placental laminin preparations, regardless of source, are
commonly contaminated with the proteins entactin and/or
collagen type IV.27 To rule these proteins out as the
mediators of sickle cell adhesion to human laminin, recombinant human
entactin and collagen type IV were immobilized and tested for their
ability to support sickle RBC adhesion. Neither protein supported the adhesion of the sickle cells beyond the background levels observed to
the BSA control (Fig 1B). Therefore, it is unlikely that either of
these contaminants is responsible for the observed adhesion of sickle
RBCs. Furthermore, an antilaminin polyclonal antibody (hLAM pAb) was
incubated on laminin-coated wells before the flow adhesion experiment.
The hLAM pAb inhibited sickle RBC adhesion to laminin by 96% (Fig 1C),
providing further evidence that laminin, not a contaminant of the
preparation, is adhesive for sickle cells.
To further characterize the adhesion of SS RBCs to laminin, sulfated
carbohydrates were tested as potential inhibitors. High molecular
weight dextran sulfate (1 mg/mL) or fucoidan (1 mg/mL) was
incubated in 1% SS RBC hematocrit for 1 hour at 37°C before the
flow adhesion assay. High molecular weight dextran sulfate (HMW DS) and
fucoidan inhibited the adhesion of SS RBCs to laminin by 87% and 80%,
respectively (data not shown), suggesting that these sulfated
carbohydrates may be useful in studies to isolate the SS RBC receptor
and characterize its interaction with laminin.
To identify the most adhesive fraction of SS RBCs, the least dense
(reticulocyte-enriched) or most dense (reticulocyte-depleted) fractions
of SS RBCs were allowed to interact with immobilized laminin under flow
conditions. The most dense SS RBC fraction was significantly more
adherent to laminin than the least dense fraction (Table 1). However,
because these fractions are not completely reticulocyte-depleted or
enriched (see Materials and Methods), we cannot firmly conclude which
age of SS RBCs is most adhesive. Interestingly, these results differ
from our previous observations of SS RBC adhesion to thrombospondin,
which supported higher levels of adhesion of least dense,
reticulocyte-enriched RBCs.23 These results suggest that
the laminin receptor on SS RBCs is more available in the denser
fraction. Furthermore, only 10% of the SS RBCs adhering to laminin had
visibly sickled morphologies (data not shown), suggesting that
extensive distortion of the SS RBC membrane is not required to expose
the laminin receptor on the SS RBC or for the adhesion to occur.
To determine if known adhesive sites of laminin mediate the adhesion of
sickle cells, peptides representing these regions34 were
tested for their activity with sickle RBCs in our flow adhesion system.
These peptides interact with a variety of cell types and are known to
retain their biological activity in peptide form in their respective
adhesion studies35-37 and when immobilized onto
plastic.38,39 The RGD sequence in the laminin chain promotes cellular attachment via integrins, IKVAV of the chain promotes neurite outgrowth, and YIGSR of the chain inhibits tumor
metastases.34 In our flow adhesion system, neither the GRGDSP, IKVAV, nor YIGSR peptides supported sickle cell adhesion beyond
background levels (Fig 2A). These peptides
(1 mmol/L) also did not inhibit the adhesion of sickle cells to laminin
when incubated with sickle RBCs before the flow adhesion assay (data
not shown). Results of this experiment suggest that none of these
regions mediates the interaction between laminin and sickle RBCs.
Moreover, laminin itself in solution did not inhibit sickle cell
adhesion to immobilized laminin (data not shown), which suggests that
adhesive sites of laminin differ in the immobilized and soluble forms.
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| Fig 2.
Sickle RBCs do not adhere to known adhesive regions of
laminin, murine EHS laminin, or human laminin-2. (A) Peptides
representing known adhesive regions of laminin (GRGDSP, IKVAV, and
YIGSR) were immobilized (1 mmol/L) and tested for their ability to
support sickle RBC adhesion. These peptides did not support sickle RBC
adhesion beyond nonspecific background levels. Adhesion to BSA and hLAM
are shown as negative and positive controls, respectively. N = number
of patients tested for each peptide. (B) SS RBCs were flowed across
immobilized laminin-2 or mouse EHS laminin. Only background levels of
sickle cells adhered to laminin-2 (hLAM-2) or mouse laminin (mLAM)
compared with human placental laminin (hLAM). Both positive and
negative controls are presented. Data shown represent the mean ± SD
from six experiments involving 3 patients.
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Human placental laminin is likely a mixture of laminin heterotrimers,
containing 1 each of the 5 known chains, 3 chains, and 2 chains. Laminin-2, or merosin, has the 2, 1, 1 configuration. When tested for its ability to support sickle cell adhesion, only background levels of sickle RBCs adhered to the immobilized protein (Fig 2B). In inhibition assays, laminin-2 also did not inhibit the
adhesion of sickle RBCs to immobilized human placental laminin (data
not shown). The lack of interaction between sickle cells and laminin-2
suggests that the 2, 1, and 1 chains are not involved in the
adhesion of sickle RBCs. Similarly, sickle cells did not adhere to
murine EHS laminin (Fig 2B). Although this may be due to disparate
regions between the mouse and human versions of the proteins, this is
less likely, because murine versions of the laminin molecules are
highly homologous40 and often behave similarly.41,42 Murine EHS laminin has been characterized
as containing the 1, 1, and 1 chains,42 further
decreasing the likelihood that the 1 or 1 chains are adhesive and
potentially eliminating the 1 chain as well. It should also be noted
that, relative to results presented below, murine EHS laminin
preparations lack the 5 chain.26
Because the greatest diversity of laminin is between its various chains, the anti- chain MoAb 4C7, which binds to the laminin 5
chain43-45 within or near the G-domain,41 was
tested for its ability to inhibit sickle cell adhesion to laminin.
Significantly less sickle cells adhered to human laminin in the
presence of the antibody (79.6% inhibition), but not in the presence
of an isotype-matched IgG2 control antibody
(Fig 3). When the commercial antibody
preparation was cleared of ascites, sickle RBC adhesion to laminin was
still inhibited by approximately 60%, whereas the ascites fluid alone
did not inhibit the adhesion (Fig 3). Finally, the 4C7 MoAb, relative
to a control MoAb, reacted strongly with immobilized laminin in an
enzyme-linked immunosorbent assay (data not shown), demonstrating that
the 4C7 epitope is present in the human placental laminin preparation
used in these studies. These results suggest that the laminin 5
chain contains an adhesive site for sickle RBCs at or near the G-domain
and are consistent with a lack of sickle cell adhesion to the laminin-2
and murine EHS laminin preparations.
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| Fig 3.
Sickle cell adhesion to laminin is inhibited by the 4C7
MoAb. Immobilized laminin was treated with the 4C7 antilaminin MoAb
(4C7 MoAb) in ascites before the 1% hematocrit flow adhesion.
Concentrations of the 4C7 IgG2 in ascites ranged from 15.0 to 37.5 µg/well. Adhesion of sickle RBCs to laminin was greatly
reduced by the 4C7 MoAb, but not by an isotype-matched IgG2
control antibody at amounts up to 37.5 µg/well or by ascites fluid
isolated from the commercial 4C7 preparation. Adhesion to human laminin
in the absence of antibody is presented as a negative control. Data
shown represent 10 experiments involving 4 patients.
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DISCUSSION |
This study provides evidence that the laminin 5 chain contains an
adhesive region for sickle RBCs. The 1, 2, 1, or 1 chains
of laminin are unlikely to account for the interaction with SS RBCs,
because laminin-2 and EHS laminin did not support sickle cell adhesion.
Furthermore, regions of laminin known to be adhesive to other cell
types are not responsible for sickle cell adhesion, because peptides
representing these regions were not active in our assay. Finally, the
4C7 MoAb, which recognizes the laminin 5 chain43-45 at
or near the G-domain,41 significantly inhibited
sickle cell adhesion to laminin. These results suggest that the laminin
5 chain contains an adhesive site within or near its G-domain to
which sickle RBCs bind under flow conditions. This conclusion is
consistent with a lack of adhesion to laminin preparations lacking the
5 chain, ie, laminin-2 and EHS laminin. The observed adhesion of
sickle cells to laminin may be a contributing factor in vascular
occlusion and the onset of the sickle cell crisis event.
Distinct topological features of the sickle RBC membrane and increased
concentrations of certain plasma components combine to make sickle
cells abnormally adherent to the endothelium.46 Sickle RBCs
may also adhere to the subendothelium when subendothelial proteins are
abnormally exposed. Endothelial damage22 and increased levels of circulating endothelial cells19-21 have both been
documented in sickle cell disease. The ECM proteins thrombospondin and
laminin are both adhesive for sickle RBCs,23,24 but laminin
supports higher levels of sickle RBC adhesion than any other reported
adhesogenic molecule. Our assay involves the examination of sickle cell
adhesion to immobilized laminin, which is most likely to mimic the
conformation of laminin in its biologically relevant form, ie,
immobilized in the ECM. With the strong correlation between levels of
sickle cell adhesion to the endothelium and clinical vaso-occlusive
severity,47 the interaction between sickle cells and
laminin may be an important determinant of crisis onset and severity.
Laminin-mediated adhesion of sickle RBCs appears to be the result of a
receptor distinct from that which mediates SS RBC binding to
thrombospondin, because the less dense, reticulocyte-enriched RBC
population is associated with a higher level of adhesion to thrombospondin.23 In contrast, the denser,
reticulocyte-depleted fraction of SS RBCs is more adhesive to laminin.
This SS RBC population contains less deformable RBCs that are more
likely to cause erythrostasis, possibly suggesting a greater likelihood
of decreased flow rates and increased opportunity to interact with the
subendothelium. Our experimental protocol precludes specific
identification of the age of cells that actually adhere, preventing a
firm conclusion of whether older cells are actually more adherent.
However, these results do suggest that the laminin receptor is more
readily available in the denser population of SS RBCs. An acidic
glycolipid has been proposed as the SS RBC receptor for both
thrombospondin and laminin.24 If so, our results suggest
that density-related conformational differences in the SS RBC membrane
make the lipid more or less adhesive for a given matrix protein.
Alternatively, the denser SS RBC may feature a separate and distinct
receptor for laminin. The protein B-CAM/LU of the lutheran blood group
antigens is also a likely receptor candidate for the adhesion of SS
RBCs to laminin, because it is uniquely adhesive to laminin in the
sickle cell disease state.48 Whichever receptor is
involved, our results with sulfated carbohydrates suggest that its
ability to bind to laminin should be inhibited by HMW DS or fucoidan.
Having been studied widely for its interactions with various cell
types, specific laminin regions have been identified as adhesive.
Peptides representing known laminin adhesive regions were tested for
their interaction with sickle RBCs. Although these peptides are
reported to support and/or inhibit the adhesion of other cell
types,35-37 we found that none had activity for sickle cells in our flow adhesion system. These results suggest that sickle
RBC adhesion to laminin is mediated by a mechanism not previously
described in other laminin adhesion studies. The adhesion also appears
to be specific to the sickle cell disease state, because normal RBCs
showed little affinity for immobilized laminin and did not adhere
beyond background levels (Fig 1A). Similarly, RBCs with high
reticulocyte counts do not adhere to laminin in the absence of the Hb S
hemoglobinopathy.24
When the MoAb 4C7 was allowed to interact with the immobilized laminin
before the introduction of sickle RBCs in the flow adhesion assay, 4C7
significantly inhibited sickle cell adhesion to laminin. Although the
antibody's epitope has not been precisely mapped, it has been shown to
be present in a basal membrane component that is expressed in
capillaries.49 Electron micrograph and immunohistochemical
studies indicate that 4C7 likely recognizes the G-domain of the laminin
5 chain.41,43-45 The inhibition of sickle cell adhesion
to laminin by 4C7 therefore suggests that the laminin 5 chain,
within or near the G-domain, is the adhesive region of laminin for
sickle cells. Because 5 forms both laminin-10 and
laminin-11,25 it is likely that these two molecules are the
laminin isoforms that are adhesive for sickle RBCs.
The laminin 5 chain is believed to have the widest expression of the
chains, with its transcripts being found in nearly all
tissues,25 making it largely available for sickle cell
interaction wherever the endothelium is damaged. The laminin 5 chain
is also found in areas of regeneration43 and is likely to
be abundant around sites of endothelial damage where higher
concentrations of plasma proteins like fibronectin also exist and
increase the adhesiveness of sickle cells.46 Furthermore,
topological orientation of the laminin molecule in the basal lamina
indicates that the G-domain is adjacent to the
endothelium,50 making it potentially available for RBCs
upon exposure of the ECM.
In our study, sickle cells did not adhere to laminin-2. This laminin
isoform (2, 1, 1) has characteristically conserved chain
regions, with homology to the other chains exceeding 80% in some
domains. However, the 2 G-domain is only 20% homologous to the 5
G-domain,51 suggesting likely differences in properties and
functions. The inability of laminin-2 to support sickle RBC adhesion
provides additional evidence that the highly conserved, known adhesive
regions of laminin are not active in sickle cell adhesion and suggest
the role of a laminin domain that is not highly conserved among the
isoforms. Of the known chains, all have similarly low levels of
homology among the G-domains. Therefore, adhesive interactions of the
G-domains are likely to be chain-specific. This may explain why sickle
cells did not adhere to the highly homologous murine laminin, which
also does not contain the 5 chain.25
It is interesting to note that, although the G-domain has been
described as responsible for most of the cell binding activity of
laminin,52 few specific regions or sequences from this
domain have been characterized for adhesion. This may be due to the
high level of variability between chain G-domains51 and
the fact that the 3,53 4,54 and
555 chains were only recently identified. Adding to the
complexity is the possible existence of novel isoforms of individual
chains, as is the case with the 3 chain.25 Moreover,
the widely used 4C7 antibody was until recently thought to interact
with the 1 G-domain,43-45 therefore causing often
conflicting data about chain expression.
Presumably, each chain has specific as well as overlapping
functions. Unique phenotypes have linked 2 to a muscular dystrophy subset56 and 3 to epidermolysis bullosa.57
This study suggests that 5 may play a role in sickle cell disease.
The recent identification of laminin 5 in an endothelial cell
line58 used widely in previous studies of sickle cell
adhesion to the endothelium9,59 suggests that laminin could
be involved in endothelial as well as subendothelial adhesion of sickle
cells. Higher levels of 5 expression in the lungs and
kidneys58 may correspond to the clinical features of acute
chest syndrome and kidney complications that are common in sickle cell
patients.2 Increased levels of 5 expression at the onset
of sexual maturity58 could possibly explain why some
patients do not experience painful crises until adolescence or early
adulthood.60 Given the amount of adhesion in vitro in the
absence of the many adhesogenic factors present in patients, sickle RBC
adhesion to laminin may prove to be an important factor in the
initiation of vaso-occlusion and the onset of crises. A therapeutic
agent designed to inhibit sickle cell adhesion to laminin may aid in
preventing vaso-occlusion, thus preserving health and improving the
quality of life of sickle cell patients.
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FOOTNOTES |
Submitted March 31, 1998;
accepted June 11, 1998.
Supported by National Institutes of Health Grants No. HL58939, HL45923,
HL45100, and RR00046.
Address reprint requests to Leslie V. Parise, PhD, Department of
Pharmacology, CB#7365, University of North Carolina, Chapel Hill, NC
27599-7365.
The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" is accordance with 18 U.S.C. section 1734 solely to indicate this fact.
| |
ACKNOWLEDGMENT |
Many thanks to David Shock for his technical assistance and to Susan
Jones, Wendy O'Kelly, B.J. Lee, Anne Criss, and the UNC Comprehensive
Sickle Cell Center for their help recruiting patients and obtaining
samples for this study.
| |
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Abstract
Introduction
Abstract