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Division of Allergy, La Jolla Institute for Allergy and Immunology, San Diego, CA 92121;
Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037;
First Department of Internal Medicine, Kumamoto University School of Medicine, Kumamoto, Japan; and
§
Department of Immunology and Immunopathology, Kagawa Medical School, Kagawa, Japan
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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, and stromal
cell-derived factor-1. Cultured human macrophages and alveolar
macrophages also migrated toward galectin-3, but not monocyte
chemotactic protein-1. Finally, galectin-3 was found to cause monocyte
accumulation in vivo in mouse air pouches. These results indicate that
galectin-3 is a novel chemoattractant for monocytes and macrophages and
suggest that the effect is mediated at least in part through a
PTX-sensitive (G protein-coupled) pathway. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Galectin-3 has been shown to function through both intracellular and extracellular actions. Related to its intracellular functions, galectin-3 has been identified as a component of heterogeneous nuclear ribonuclear protein (hnRNP) (26), a factor in pre-mRNA splicing (27), and has been found to control cell cycle (28) and prevent T cell apoptosis; the latter is probably mediated through interaction with the Bcl-2 family members (29, 30). On the other hand, this protein, which is secreted from monocytes/macrophages (31, 32) and epithelial cells (33, 34), has also been demonstrated to function as an extracellular molecule in activating various types of cells, including monocytes/macrophages (13), mast cells (35, 36), neutrophils (37), and lymphocytes (15, 38). Galectin-3 has been shown to mediate cell-cell and cell-extracellular matrix interactions (39, 40, 41).
Recently, it has been shown that galectin-9, another member of the galectin family with two CRDs, is a selective chemoattractant for eosinophils (42). The activity is dependent on both CRDs, suggesting that cross-linking of cell surface molecules is involved in the chemoattraction (43). Galectin-3 is known to form dimers through the amino-terminal nonlectin domain and thus has the potential to cross-link appropriate cell surface glycoproteins (9). Therefore, it is conceivable that galectin-3 also has similar activity for certain cell types.
In the present study we focused on an extracellular function of galectin-3 on monocyte/macrophage, because 1) galectin-3 is known to bind to the cell surfaces of monocytes/macrophages (44), and 2) high expression of galectin-3 is seen in human and rat lungs (45, 46), where macrophages are one of the dominant cell types. Moreover, we recently found that macrophage recruitment in peritonitis is attenuated in galectin-3-deficient mice (47). As inflammatory macrophages are known to result from the recruitment of peripheral blood monocytes, these facts led us to examine whether galectin-3 attracts monocytes and macrophages. We found that galectin-3 induced migration of monocytes and macrophages both in vitro and in vivo through a pertussis toxin (PTX)-sensitive pathway. Our present study suggests a novel role for galectin-3 in inflammation and malignancy through its ability to recruit inflammatory cells and APCs into the affected sites to modify the biological responses. These results together with the finding that galectin-9 is a potent eosinophil chemoattractant suggest that galectins are a new class of chemoattractants.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Recombinant human galectin-3 (48), the C-terminal
domain fragment of galectin-3 (galectin-3C) (29), a mouse
mAb against galectin-3 (B2C10) (49), and mouse monoclonal
anti-DNP IgG1 (50) were prepared as described
previously. Recombinant MCP-1, MIP-1, and SDF-1 were obtained
from PeproTech (Rocky Hill, NJ). Indo-1/AM was purchased from Molecular
Probes (Eugene, OR). HBSS and RPMI 1640 were purchased from Life
Technologies (Grand Island, NY). Ficoll Paque and Percoll solution were
obtained from Amersham Pharmacia Biotech (Uppsala, Sweden). Unless
otherwise stated, all other reagents were purchased from Sigma (St.
Louis, MO).
Preparation of human monocytes
Human monocytes were purified from venous blood of normal volunteers essentially as described previously (51). In brief, after erythrocytes were sedimented by addition of 6% dextran saline solution (1 part to 5 parts heparinized blood), and the leukocytes were collected, washed twice, and resuspended in Ca2+ and Mg2+-free HBSS containing 5% autologous serum. Mononuclear cells were acquired by centrifugation of the leukocyte suspension on Ficoll Paque at 800 x g for 15 min. The cells were resuspended in RPMI 1640 containing 10% autologous serum and allowed to adhere to sterile tissue culture plates for 30 min in a humidified incubator at 5% CO2 and 37°C. After incubation, nonadherent cells were removed by washing the plates three times with PBS at 37°C. Greater than 98% of the adherent cells showed the characteristic appearance of monocytes when examined by light microscopy following Wright staining or neutral red staining. To detach and harvest the adhered monocytes, 1 mM EDTA-PBS containing 5% serum was added, and the plates were incubated on ice for 30 min. The monocytes were washed twice with HBSS and resuspended in RPMI 1640 with 0.1% autologous serum for the migration assay. The viability of monocytes was determined by trypan blue exclusion and was >98%. In some experiments monocytes were purified according to another method using a Percoll discontinuous gradient described previously (52). No difference was noted in the purity and viability of the cells prepared by these two different methods.
Preparation of human cultured peripheral blood macrophages and alveolar macrophages
Human macrophages were obtained by culturing peripheral blood monocytes in vitro for 7 days as previously described (53). Human alveolar macrophages were obtained from bronchoalveolar lavage fluid according to a previously described protocol (54). The purity of the macrophages was >90%, and the viability was >99%.
Migration assay in vitro
Monocyte migration was examined by using 96-well micro-Boyden chambers with 5 µm pore size filters (Neuro Probe, Gaithersburg, MD) as described previously (55, 56). Briefly, after the indicated concentrations of galectin-3 or MCP-1, in RPMI 1640 were applied to the lower chambers, purified monocyte suspensions (2.5–5.0 x 104/well) were applied to the upper chambers. After incubation of the chambers for 1 h in a humidified incubator at 5% CO2 and 37°C, the filters were washed once with PBS and processed with Wright stain. The number of monocytes on the bottom side of the filters was counted in 5–10 high power fields. Monocyte migration was calculated from the average numbers of the counted cells and expressed as the percentage of input cells in a well.
In assays using inhibitory reagents, the purified monocytes were pretreated with or without the indicated concentrations of B2C10 (49) or anti-DNP IgG1 (50) as an isotype-matched control mAb, galectin-3C, or PTX at 37°C for 30 min. Then the cells were applied to the upper chambers in the presence of these inhibitors at the same concentrations used in the pretreatment. Galectin-3 or MCP-1 was applied to the lower chambers as described above. In the assays using lactose and sucrose, the sugars were added to the lower chambers at the initiation of the migration assay.
Migration assay in vivo
The mouse air pouch experiments were performed according to a method described previously (57). Briefly, an air pouch was induced on the back of BALB/c mice by injecting 3 ml of air intradermally three times (one time each on 6, 4, and 2 days before the experiments). Then, 1 ml of 0.9% sodium chloride (USP grade saline; Baxter Healthcare, Deerfield, IL) containing 1 µM galectin-3 was injected into the pouch. As positive and negative controls, 100 ng/ml of recombinant MCP-1 and diluent only, respectively, were injected. Four hours afterward, recruited cells were recovered by gently lavaging the pouch with 1 ml of PBS containing 1 mM EDTA. Cell number was determined, and the distribution of leukocyte types was analyzed after cytospin preparation and Wright staining.
Measurement of Ca2+ influx in monocytes
Intracellular concentrations of Ca2+ were measured using indo-1/AM according to a previously described method (58). Purified monocytes were resuspended in HBSS containing 1 mM Ca2+, 1 mM Mg2+, and 5% autologous serum and incubated with 10 mM indo-1/AM for 45 min at 37°C. The cells were washed once and resuspended in the same buffer, and stimuli and inhibitors were added at the time points specified in the figure legends. Intracellular Ca2+ concentration was measured by monitoring light emission at 405 and 485 nm to an excitation wavelength of 355 nm, using an AMINCO-Bowman series 2 luminescence spectrometer (Rochester, NY).
Data analysis
Data are summarized as the mean ± SD. The statistical examination of the results was performed by the variance analysis using Fisher’s protected least significant difference test for multiple comparisons. The analysis of the results from the mouse air pouch experiments was conducted with the Mann-Whitney test. Values of p < 0.05 were considered significant.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Using a micro-Boyden chamber assay, we found that human
recombinant galectin-3 induced monocyte migration in a dose-dependent
manner. Galectin-3 significantly increased monocyte migration at
concentrations >100 nM compared with diluent and showed a bell-shaped
dose-dependent pattern like many chemoattractants (control, 3.54
± 2.2%; 100 nM, 6.25 ± 1.3%; 300 nM, 9.8 ± 0.33%; 1
µM, 12.4 ± 1.2%; 3 µM, 21.6 ± 0.071%; 10 µM,
17.04 ± 0.28%, p < 0.05 vs control;
n = 4 experiments; Fig. 1
). While the difference in the effect
between lower concentrations of galectin and control was not
statistically significant in these initial experiments, in a number of
subsequent ones, 10 nM galectin-3 also significantly increased monocyte
migration (control, 4.26 ± 1.3%; 10 nM, 7.01 ± 2.1%;
p < 0.001 vs control; n = 21). The
effect of 3 µM galectin-3 on monocyte migration was greater than that
of human recombinant MCP-1, a strong chemoattractant for monocytes
(59), at 100 ng/ml (11.6 nM; Fig. 1), which was determined
in dose-response experiments to be the concentration that induced
maximum monocyte migration in this assay (data not shown).
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). B2C10 did
not affect MCP-1-induced monocyte migration significantly under
identical conditions (data not shown). These results indicate that
exogenous galectin-3 induces migration of human monocytes in vitro.
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The observed monocyte migration induced by galectin-3 could be due
to a chemotactic or chemokinetic effect. The former means that the cell
migration is strictly a consequence of the lectin’s chemoattraction
activity, whereas the latter implies that the cell migration is due to
activation of the cells by the lectin that results in an enhanced
movement of the cells in all directions. To differentiate between these
two possibilities, we performed a checkerboard analysis. Various
concentrations of galectin-3 were applied to the upper and/or lower
chambers, and monocyte migration was examined. As shown in Table I and Fig. 3, when 10 or 100 nM galectin-3 was used,
no significant difference in monocyte migration was observed regardless
of whether the protein was added to the lower chambers or to both
chambers. In contrast, when 1 µM galectin-3 was added to both
chambers, no significant increase in monocyte migration over the
background was observed. These results indicate that the effect of
galectin-3 in vitro is chemokinetic at low concentrations (10 and 100
nM), but chemotactic at high concentrations (1 µM).
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As mentioned above, galectin-3 is composed of a C-terminal lectin
domain and an N-terminal nonlectin part. To determine whether the
chemoattractant activity of galectin-3 is dependent on its lectin
properties, we tested the effect of saccharides on the lectin’s
induction of monocyte migration. As shown in Fig. 4A, 5 mM lactose significantly
decreased monocyte migration induced by 10 nM, 100 nM, and 1 µM
galectin-3 by 63.8, 71.5, and 57.6%, respectively
(p < 0.05; n = 3). Similarly,
10 mM lactose significantly inhibited the migration by 78, 74.1, and
71.1%, respectively (p < 0.05;
n = 3). These concentrations of lactose did not affect
the monocyte migration induced by MCP-1 (100 ng/ml MCP-1, 20.0 ±
3.2%; 100 ng/ml MCP-1 plus 10 mM lactose, 20.2 ± 2.1%;
n = 3). As a negative control, we also tested the
effect of sucrose, which dose not bind to galectin-3. As shown in Fig. 4B, sucrose had no significant effect on monocyte migration.
These results indicate that the C-terminal lectin domain of galectin-3
is involved in the induction of monocyte migration.
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, 1
µM galectin-3C alone did not have
any chemokinetic effect on monocytes, but it significantly inhibited
cell migration induced by 100 nM and 1 µM galectin-3 by 77.4 and
45.0%, respectively (p < 0.05;
n = 3). Galectin-3C pretreated at 100°C showed no
effect on galectin-3-induced monocyte migration (data not shown). We
also tested the effect of 100 nM galectin-3C, but no influence on
monocyte migration was observed (Fig. 5). These results further confirm
the involvement of the lectin domain in the chemoattractant activity
and suggest that the N-terminal domain is also necessary for this
activity.
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We tested the possibility that G proteins might be involved in
galectin-3-induced monocyte migration using the inhibitor PTX, because
it is well known that many chemoattractants, including all chemokines,
use G protein-coupled receptors to transduce signals into the cell
(60). Before the experiments, we confirmed that 1 µg/ml
of PTX did not decrease the viability of monocytes (data not shown). We
found that PTX decreased monocyte migration induced by 1 µM
galectin-3 by 91.2% (p < 0.01;
n = 5; Fig. 6A). However, PTX only
partially inhibited monocyte migration induced by 10 or 100 nM
galectin-3 (p = 0.8501 and 0.3093,
respectively; n = 5). In contrast, 1 µg/ml of PTX
significantly inhibited monocyte migration induced by MCP-1 at all
concentrations examined (Fig. 6B). These results suggest
that a PTX-sensitive G protein-coupled receptor(s) is involved in
monocyte migration induced by galectin-3.
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As mentioned above, galectin-3 can dimerize and cross-link cell
surface receptors. This suggests that the reason why galectin-3 is
chemotactic is because it is able to activate chemokine receptors.
Therefore, to further analyze galectin-3-mediated signaling, we tested
the effect of this lectin to induce a Ca2+ influx
in monocytes, because many chemoattractants are known to cause
Ca2+ influx. We found that 1 µM galectin-3, but
not lower concentrations, induced a Ca2+ influx
in human monocytes similar to MCP-1 (Fig. 7, A and B),
although the extent of the Ca2+ influx caused by
the lectin was lower than that caused by the chemokine in all three
separate experiments. We found that heat-inactivated galectin-3 did not
produce any response (data not shown). The specificity of this activity
was also demonstrated by the complete inhibition of galectin-3-induced,
but not MCP-1-induced, Ca2+ influx by 5 mM
lactose, but not sucrose (Fig. 7, C and D).
Furthermore, both the galectin-3- and the MCP-1-induced
Ca2+ influx were blocked by PTX (Fig. 7, E and F). These results indicate that galectin-3
causes a Ca2+ influx, which is probably mediated
by a PTX-sensitive G protein coupled receptor(s).
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Among various chemoattractants, the monocyte/macrophage-reactive
chemokines, including MCP-1, MIP-1, and SDF-1, are known to cause
a Ca2+ influx in the cells (61, 62, 63)
by binding to their receptors, such as CCR2/9, CCR1/5/9, and CXCR-4,
respectively, all of which are coupled with PTX-sensitive G proteins
(60, 64, 65). To determine the possibility that galectin-3
interacts with these receptors to transduce activation signal(s) into
monocytes, we performed Ca2+ influx experiments
to study cross-desensitization. This method is known to be useful in
identifying the usage of the chemoattractant receptors, although
cross-desensitization occurs at multiple levels and can affect signals
mediated by other receptors (66, 67). We found that all
these chemokines (100 ng/ml) induced a Ca2+
influx in human monocytes (Fig. 8, A, C, and E). We also observed that
such responses were desensitized by pretreatment with the same, but not
other, chemokines (data not shown), consistent with previous results
from other investigators (61, 62, 63). We found, however, that
there was no cross-desensitization between galectin-3 and any of the
above-mentioned monocyte-reactive chemokines (Fig. 8). These results
suggest that galectin-3 does not interact with any of these presently
known chemokine receptors expressed on monocytes for signal
transmission into the cell.
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Unlike monocytes, there are fewer chemokines that have been shown
to attract mature macrophages (65). To determine the
effect of galectin-3 on mature macrophages, we used human macrophages
obtained from culturing peripheral blood monocytes as well as alveolar
macrophages. A previous study showed that cultured human macrophages do
not express a detectable amount of CCR2 and do not respond to its
ligand, MCP-1 (53), which we also confirmed (Fig. 9). In contrast, we found that galectin-3
induced macrophage migration in a dose-dependent manner, and 1 µM
galectin-3 enhanced migration by 190% over that induced by the control
medium (p < 0.05; n = 3; Fig. 9). Similarly, human alveolar macrophages migrated toward galectin-3 in
two separate experiments (Fig. 10). In
these experiments, bell-shaped dose-response curves were obtained,
which is commonly observed for many chemokines. In contrast, MCP-1 had
no effect (Fig. 10, Expt 1) or a negligible effect (Fig. 10, Expt 2) on
macrophage migration. These results indicate that galectin-3, but not
MCP-1, is a chemoattractant for macrophages. The results also
corroborate the conclusion made above that the signaling pathway
induced by galectin-3 is not mediated through CCR2.
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Lastly, we examined the effect of galectin-3 on cell recruitment
into mouse air pouches to determine whether galectin-3 induces
migration of cells in vivo. As shown in Fig. 11, galectin-3 increased the numbers of
monocytes, neutrophils, and eosinophils in the air pouch by 15.7, 5.30,
and 4.63 times, respectively, over those induced by vehicle (saline)
only (p < 0.05; n = 6). In
contrast, the number of lymphocytes was not augmented significantly by
the treatment (p = 0.309). We tested galectin-3
samples pretreated at 100°C for 5 min and found that these samples
caused a background level of inflammatory cell accumulation comparable
to that induced by saline (data not shown). These results indicate that
galectin-3 can recruit monocytes in vivo.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The finding that galectin-3 is a chemoattractant for macrophages in addition to monocytes is noteworthy, because unlike monocytes, there are fewer chemokines that have been shown to attract mature macrophages (65). The major monocyte chemoattractant, MCP-1, for example, is inactive in this respect. Therefore, it is possible that galectin-3 is one of the major factors involved in the influx of macrophages to inflammatory sites. Based on the results from the present study, we propose that galectin-3 produced by epithelial cells, a major source of this lectin, can contribute to the attraction of monocytes and macrophages during inflammation. Because monocytes and macrophages also produce galectin-3, this lectin may mediate a continued influx of these cell types once the inflammatory process is initiated. Recently, we found that galectin-3-deficient mice developed significantly reduced numbers of peritoneal macrophages compared with wild-type mice when treated with thioglycolate i.p. (47). These results are highly consistent with the present findings, and together they suggest that galectin-3 released by the peritoneal cells in thioglycolate-treated mice is responsible at least in part for recruiting monocytes and macrophages to the peritoneal cavity, and that galectin-3-deficient mice exhibit a lower macrophage response due to the absence of this chemoattractant.
It has been shown previously that galectin-3 can activate various cell types, including induction of superoxide production by monocytes/macrophages (13). Although the precise mechanisms remain to be determined, these activities are probably related to the dimerization or oligomerization of galectin-3 through intermolecular interactions involving the amino-terminal domain (48). The lectin thereby becomes bivalent or multivalent functionally and capable of activating cells by effectively cross-linking cell surface glycoproteins (3, 4, 5, 6, 7). We believe that this process also contributes to the monocyte chemoattractant activity of galectin-3, and this possibility is supported by our finding that both the N-terminal and C-terminal domains of galectin-3 are required for this activity. However, an unusual feature of galectin-3’s chemoattractant activity is that the response is both qualitatively and quantitatively dependent on the concentration of the lectin. First, galectin-3 appears to be chemokinetic at low concentrations, but chemotactic at high concentrations. One possible explanation is that galectin-3 at high concentrations can cause cell aggregation, and thus in the checkerboard analysis, when galectin-3 was added to the upper chambers together with the cells, the cells were prevented from migrating toward the lower chambers because they were aggregated. Therefore, it might be possible that galectin-3 is actually chemokinetic for monocytes at both high and low concentrations.
However, only monocyte migration induced by high concentrations of galectin-3 was completely inhibited by PTX. Also, only high concentrations of galectin-3 caused a Ca2+ influx in monocytes, and this occurred through a PTX-sensitive mechanism(s). The most likely explanation for these findings is that galectin-3 binds to and activates different (or different sets of) cell surface molecules depending on its concentration. At lower concentrations, it preferentially binds to glycoproteins that interact with the lectin relatively strongly, while only after reaching a certain threshold concentration does it begin to recognize other cell surface glycoproteins that interact with the lectin relatively weakly. Our data suggest that the latter include a PTX-sensitive, G protein-coupled receptor(s). Galectin-3 has already been shown to bind to a number of different cell surface glycoproteins on macrophages (68), and based on a recent study with galectin-1 (69), it is likely that the lectin can cause segregation of these different glycoproteins. It is entirely possible that the lectin binds to these different glycoproteins with variable affinity, because they are differentially glycosylated, and the lectin exhibits a fine specificity for oligosaccharides (45, 46, 70).
Relatively high concentrations of galectin-3 are needed for the
demonstration of optimal chemoattractant activity. The situation is
analogous to other activities demonstrated for this lectin previously,
such as activation of inflammatory cells (13, 35, 37), and
is probably related to the concentrations that are required for the
dimerization or oligomerization of the lectin to take place. However,
galectin-3 is known to exist at relatively high concentrations in the
cytosol of many cell types (e.g., 5 µM in a human colon
adenocarcinoma cell line, T84 (71)). Therefore, a high
local concentration of the lectin may be achieved when there is a burst
release of the protein from these cells. In fact, galectin-3 has been
found to be present in significant amounts in biological fluids. For
example, the concentrations of galectin-3 in bronchoalveolar lavage
fluid from mice with airway inflammation were >20 nM (our unpublished
observation). Considering the dilution factor introduced in obtaining
the lavage fluid, it is easily conceivable that the initial local
concentrations of the lectin are in the micromolar range. On the other
hand, the effective concentrations of galectin-3 for attracting
alveolar macrophages are much lower (Fig. 10), approaching those
typically found for many chemokines. It is possible that the putative
receptor for galectin-3 on these cells either exists in higher numbers
or interacts with the lectin more strongly. Alternatively, the putative
receptor on these cells transmits signals more effectively upon
interacting with the lectin.
Galectin-3 probably activates PTX-sensitive, G protein-coupled
receptors similar to those recognized by many known chemokines
(60, 64). This lectin does not have significant sequence
similarity with any of these chemokines, and thus it appears unlikely
that it recognizes these receptors through protein-protein
interactions, but it could do so via lectin-carbohydrate interactions.
Chemokine receptors expressed on monocytes include CCR-1, CCR-2, CCR-5,
and CXCR-4 (60, 64). However, we found that there was no
cross-desensitization between galectin-3 and any of the
monocyte-reactive chemokines that use these receptors, including MCP-1
for CCR-2, MIP-1 for CCR-1 and CCR-5, and SDF-1 for CXCR-4
(60, 61, 62, 63, 64). We also did not detect interactions between
galectin-3 and these four chemokine receptors by immunoprecipitation
and immunoblotting using specific Abs (data not shown). It has been
reported that CCR-3 may also be expressed on human monocytes and
macrophages (53). However, we did not analyze the usage of
this receptor, because we observed that galectin-3 does not attract
eosinophils (which are known to express CCR-3) in vitro (our
unpublished data), and these cells are known to selectively express
CCR-3, suggesting that galectin-3 does not act on this receptor.
Therefore, in this study we could not determine the exact receptor(s)
for galectin-3, but it is not any of the known receptors, such as
CCR-1, CCR-2, CCR-3, CCR-5, and CXCR-4.
Other types of chemoattractant receptors, including those for fMLP, platelet-activating factor, leukotrienes, and C5a, could mediate the effects of galectin-3. Galectin-3 is also known to recognize CD11b, LAMP1 and -2, Mac-3, and CD98 on thioglycolate-stimulated mouse peritoneal macrophages (68). Stimulation and/or cross-linking of CD11b and CD98 could enhance adhesion and transendothelial migration of monocytes (72, 73). However, we tested the effects of anit-CD11b and anti-CD98 Abs in our monocyte migration assay and found that they do not inhibit galectin-3’s activity (data not shown), suggesting that these two do not mediate the observed chemotaxis function of galectin-3.
It should be noted that while galectin-3 is likely to bind to a number
of different cell types through lectin-carbohydrate interactions, its
chemoattractant activity is cell type-specific, as it does not induce
migration of lymphocytes and eosinophils in vitro (our unpublished
data) and lymphocytes in vivo (Fig. 11). This selectivity could be
explained by the differential expression of the putative galectin-3
receptor on different cell types. For example, galectin-3 is known to
cause a Ca2+ influx in Jurkat T cells, but the
effect was sustained and insensitive to PTX (38), in
contrast to the effect in monocytes (Fig. 7). Thus, this lectin can use
different receptors on different cell types, resulting in the
activation of selected types of cells, or cause a similar effect(s) on
different types of cells by alternative pathways. On the other hand,
although this study was focused on monocytes/macrophages, preliminary
in vitro data suggest that galectin-3 may be a chemoattractant for
neutrophils as well (data not shown). We also found that lower
concentrations of this lectin were required for maximum migration of
neutrophils compared with monocytes. In addition, galectin-3-induced
recruitment of neutrophils in the mouse air pouch experiments (Fig. 11). The neutrophil chemoattractant activity of galectin-3 is also
consistent with the results obtained from studies of
galectin-3-deficient mice by other investigators (74), who
noted that galectin-3 deficiency results in a significantly lower
degree of neutrophil response in the peritoneal cavity following
thioglycolate stimulation. Finally, galectin-3 was shown to also
attract eosinophils in vivo (Fig. 11). However, it is possible that the
lectin stimulates certain resident or recruited cells, such as
monocytes, macrophages, and neutrophils, to release eosinophil
chemoattractant(s). This proposed mechanism appears to be operative for
MCP-1 also, which is known not to be an eosinophil chemoattractant, but
appears to recruit this cell type into the air pouches (Fig. 11).
In conclusion, galectin-3 is a novel chemoattractant for monocytes and macrophages as well as neutrophils by a PTX-sensitive pathway, which is probably different from that used by the presently known monocyte-reactive chemokines. The expression of this lectin at the sites of inflammation and malignancy may modify the biological responses through the control of recruitment and activation of monocytes/macrophages, which act as inflammatory effector cells and APCs. It is interesting to note that galectin-3 has multiple functions in addition to chemoattraction, much like many of the chemokines, which possess a broad spectrum of other activities (75, 76, 77). Finally, our present work together with the recent studies by other investigators on the eosinophil chemoattractant activity of galectin-9 (42) suggest the possibility that galectins in general could act as cross-linkers of cell surface molecules, through their structurally conserved CRDs, to induce chemoattraction. While the underlying mechanisms await elucidation, a picture that emerges is that the galectin family is a new class of chemoattractants.
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Fu-Tong Liu, La Jolla Institute for Allergy and Immunology, Division of Allergy, 10355 Science Center Drive, San Diego, CA 92121.
3 Abbreviations used in this paper: CRD, carbohydrate-recognition domain; galectin-3C, the C-terminal domain fragment of galectin-3; MCP-1, monocyte chemotactic protein-1; MIP-1, macrophage inflammatory protein-1; PTX, pertussis toxin; SDF-1, stromal cell-derived factor-1.
Received for publication January 11, 2000. Accepted for publication May 31, 2000.
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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 A. Karlsson, K. Christenson, M. Matlak, A. Bjorstad, K. L Brown, E. Telemo, E. Salomonsson, H. Leffler, and J. Bylund Galectin-3 functions as an opsonin and enhances the macrophage clearance of apoptotic neutrophils Glycobiology, January 1, 2009; 19(1): 16 - 20. [Abstract] [Full Text] [PDF]
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Y. Li, M. Komai-Koma, D. S. Gilchrist, D. K. Hsu, F.-T. Liu, T. Springall, and D. Xu Galectin-3 Is a Negative Regulator of Lipopolysaccharide-Mediated Inflammation J. Immunol., August 15, 2008; 181(4): 2781 - 2789. [Abstract] [Full Text] [PDF]
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 S. R. Stowell, C. M. Arthur, K. A. Slanina, J. R. Horton, D. F. Smith, and R. D. Cummings Dimeric Galectin-8 Induces Phosphatidylserine Exposure in Leukocytes through Polylactosamine Recognition by the C-terminal Domain J. Biol. Chem., July 18, 2008; 283(29): 20547 - 20559. [Abstract] [Full Text] [PDF]
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S. R. Stowell, C. M. Arthur, P. Mehta, K. A. Slanina, O. Blixt, H. Leffler, D. F. Smith, and R. D. Cummings Galectin-1, -2, and -3 Exhibit Differential Recognition of Sialylated Glycans and Blood Group Antigens J. Biol. Chem., April 11, 2008; 283(15): 10109 - 10123. [Abstract] [Full Text] [PDF]
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 M. Papaspyridonos, E. McNeill, J. P. de Bono, A. Smith, K. G. Burnand, K. M. Channon, and D. R. Greaves Galectin-3 Is an Amplifier of Inflammation in Atherosclerotic Plaque Progression Through Macrophage Activation And Monocyte Chemoattraction Arterioscler. Thromb. Vasc. Biol., March 1, 2008; 28(3): 433 - 440. [Abstract] [Full Text] [PDF]
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J. Nieminen, C. St-Pierre, P. Bhaumik, F. Poirier, and S. Sato Role of Galectin-3 in Leukocyte Recruitment in a Murine Model of Lung Infection by Streptococcus pneumoniae J. Immunol., February 15, 2008; 180(4): 2466 - 2473. [Abstract] [Full Text] [PDF]
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A. C. MacKinnon, S. L. Farnworth, P. S. Hodkinson, N. C. Henderson, K. M. Atkinson, H. Leffler, U. J. Nilsson, C. Haslett, S. J. Forbes, and T. Sethi Regulation of Alternative Macrophage Activation by Galectin-3 J. Immunol., February 15, 2008; 180(4): 2650 - 2658. [Abstract] [Full Text] [PDF]
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S. L. Farnworth, N. C. Henderson, A. C. MacKinnon, K. M. Atkinson, T. Wilkinson, K. Dhaliwal, K. Hayashi, A. J. Simpson, A. G. Rossi, C. Haslett, et al. Galectin-3 Reduces the Severity of Pneumococcal Pneumonia by Augmenting Neutrophil Function Am. J. Pathol., February 1, 2008; 172(2): 395 - 405. [Abstract] [Full Text] [PDF]
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M. Nachtigal, A. Ghaffar, and E. P. Mayer Galectin-3 Gene Inactivation Reduces Atherosclerotic Lesions and Adventitial Inflammation in ApoE-Deficient Mice Am. J. Pathol., January 1, 2008; 172(1): 247 - 255. [Abstract] [Full Text] [PDF]
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S. P. Rao, Z. Wang, R. I. Zuberi, L. Sikora, N. S. Bahaie, B. L. Zuraw, F.-T. Liu, and P. Sriramarao Galectin-3 Functions as an Adhesion Molecule to Support Eosinophil Rolling and Adhesion under Conditions of Flow J. Immunol., December 1, 2007; 179(11): 7800 - 7807. [Abstract] [Full Text] [PDF]
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 L. Breuilh, F. Vanhoutte, J. Fontaine, C. M. W. van Stijn, I. Tillie-Leblond, M. Capron, C. Faveeuw, T. Jouault, I. van Die, P. Gosset, et al. Galectin-3 Modulates Immune and Inflammatory Responses during Helminthic Infection: Impact of Galectin-3 Deficiency on the Functions of Dendritic Cells Infect. Immun., November 1, 2007; 75(11): 5148 - 5157. [Abstract] [Full Text] [PDF]
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 F. H.M. de Melo, D. Butera, R. S. Medeiros, L. N. d. S. Andrade, S. Nonogaki, F. A. Soares, R. A. Alvarez, A. M. Moura da Silva, and R. Chammas Biological Applications of a Chimeric Probe for the Assessment of Galectin-3 Ligands J. Histochem. Cytochem., October 1, 2007; 55(10): 1015 - 1026. [Abstract] [Full Text] [PDF]
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 F. L. Oliveira, P. Frazao, R. Chammas, D. K. Hsu, F. T. Liu, R. Borojevic, C. M. Takiya, and M. C. El-Cheikh Kinetics of mobilization and differentiation of lymphohematopoietic cells during experimental murine schistosomiasis in galectin-3 / mice J. Leukoc. Biol., August 1, 2007; 82(2): 300 - 310. [Abstract] [Full Text] [PDF]
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J. Nieminen, A. Kuno, J. Hirabayashi, and S. Sato Visualization of Galectin-3 Oligomerization on the Surface of Neutrophils and Endothelial Cells Using Fluorescence Resonance Energy Transfer J. Biol. Chem., January 12, 2007; 282(2): 1374 - 1383. [Abstract] [Full Text] [PDF]
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 E Lippert, W Falk, F Bataille, T Kaehne, M Naumann, M Goeke, H Herfarth, J Schoelmerich, and G Rogler Soluble galectin-3 is a strong, colonic epithelial-cell-derived, lamina propria fibroblast-stimulating factor Gut, January 1, 2007; 56(1): 43 - 51. [Abstract] [Full Text] [PDF]
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P. Barrionuevo, M. Beigier-Bompadre, J. M. Ilarregui, M. A. Toscano, G. A. Bianco, M. A. Isturiz, and G. A. Rabinovich A Novel Function for Galectin-1 at the Crossroad of Innate and Adaptive Immunity: Galectin-1 Regulates Monocyte/Macrophage Physiology through a Nonapoptotic ERK-Dependent Pathway J. Immunol., January 1, 2007; 178(1): 436 - 445. [Abstract] [Full Text] [PDF]
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 M. Nasirikenari, B. H. Segal, J. R. Ostberg, A. Urbasic, and J. T. Lau Altered granulopoietic profile and exaggerated acute neutrophilic inflammation in mice with targeted deficiency in the sialyltransferase ST6Gal I Blood, November 15, 2006; 108(10): 3397 - 3405. [Abstract] [Full Text] [PDF]
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R. Jin, A. Greenwald, M. D. Peterson, and T. K. Waddell Human Monocytes Recognize Porcine Endothelium via the Interaction of Galectin 3 and {alpha}-GAL J. Immunol., July 15, 2006; 177(2): 1289 - 1295. [Abstract] [Full Text] [PDF]
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 S. Cinti, G. Mitchell, G. Barbatelli, I. Murano, E. Ceresi, E. Faloia, S. Wang, M. Fortier, A. S. Greenberg, and M. S. Obin Adipocyte death defines macrophage localization and function in adipose tissue of obese mice and humans J. Lipid Res., November 1, 2005; 46(11): 2347 - 2355. [Abstract] [Full Text] [PDF]
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J. Nieminen, C. St-Pierre, and S. Sato Galectin-3 interacts with naive and primed neutrophils, inducing innate immune responses J. Leukoc. Biol., November 1, 2005; 78(5): 1127 - 1135. [Abstract] [Full Text] [PDF]
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 J M Ilarregui, G A Bianco, M A Toscano, and G A Rabinovich The coming of age of galectins as immunomodulatory agents: impact of these carbohydrate binding proteins in T cell physiology and chronic inflammatory disorders Ann Rheum Dis, November 1, 2005; 64(suppl_4): iv96 - iv103. [Abstract] [Full Text] [PDF]
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S.-Y. Dai, R. Nakagawa, A. Itoh, H. Murakami, Y. Kashio, H. Abe, S. Katoh, K. Kontani, M. Kihara, S.-L. Zhang, et al. Galectin-9 Induces Maturation of Human Monocyte-Derived Dendritic Cells J. Immunol., September 1, 2005; 175(5): 2974 - 2981. [Abstract] [Full Text] [PDF]
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A. N. Moreno, G. Pereira-da-Silva, C. Oliver, M. C. Jamur, A. Panunto-Castelo, and M. C. Roque-Barreira The Macrophage-derived Lectin, MNCF, Activates Neutrophil Migration through a Pertussis Toxin-sensitive Pathway J. Histochem. Cytochem., June 1, 2005; 53(6): 715 - 723. [Abstract] [Full Text] [PDF]
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 N. Ortega, D. J. Behonick, C. Colnot, D. N.W. Cooper, and Z. Werb Galectin-3 Is a Downstream Regulator of Matrix Metalloproteinase-9 Function during Endochondral Bone Formation Mol. Biol. Cell, June 1, 2005; 16(6): 3028 - 3039. [Abstract] [Full Text] [PDF]
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G. C. Fernandez, J. M. Ilarregui, C. J. Rubel, M. A. Toscano, S. A. Gomez, M. Beigier Bompadre, M. A. Isturiz, G. A. Rabinovich, and M. S. Palermo Galectin-3 and soluble fibrinogen act in concert to modulate neutrophil activation and survival: involvement of alternative MAPK pathways Glycobiology, May 1, 2005; 15(5): 519 - 527. [Abstract] [Full Text] [PDF]
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 M. von Wolff, X. Wang, H.-J. Gabius, and T. Strowitzki Galectin fingerprinting in human endometrium and decidua during the menstrual cycle and in early gestation Mol. Hum. Reprod., March 1, 2005; 11(3): 189 - 194. [Abstract] [Full Text] [PDF]
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R. I. Zuberi, D. K. Hsu, O. Kalayci, H.-Y. Chen, H. K. Sheldon, L. Yu, J. R. Apgar, T. Kawakami, C. M. Lilly, and F.-T. Liu Critical Role for Galectin-3 in Airway Inflammation and Bronchial Hyperresponsiveness in a Murine Model of Asthma Am. J. Pathol., December 1, 2004; 165(6): 2045 - 2053. [Abstract] [Full Text] [PDF]
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Y. Y. Kleshchenko, T. N. Moody, V. A. Furtak, J. Ochieng, M. F. Lima, and F. Villalta Human Galectin-3 Promotes Trypanosoma cruzi Adhesion to Human Coronary Artery Smooth Muscle Cells Infect. Immun., November 1, 2004; 72(11): 6717 - 6721. [Abstract] [Full Text] [PDF]
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 D. Houzelstein, I. R. Goncalves, A. J. Fadden, S. S. Sidhu, D. N. W. Cooper, K. Drickamer, H. Leffler, and F. Poirier Phylogenetic Analysis of the Vertebrate Galectin Family Mol. Biol. Evol., July 1, 2004; 21(7): 1177 - 1187. [Abstract] [Full Text] [PDF]
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T. Shimaoka, T. Nakayama, K. Hieshima, N. Kume, N. Fukumoto, M. Minami, K. Hayashida, T. Kita, O. Yoshie, and S. Yonehara Chemokines Generally Exhibit Scavenger Receptor Activity through Their Receptor-binding Domain J. Biol. Chem., June 25, 2004; 279(26): 26807 - 26810. [Abstract] [Full Text] [PDF]
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W. Savino, D. A. Mendes-da-Cruz, S. Smaniotto, E. Silva-Monteiro, and D. M. S. Villa-Verde Molecular mechanisms governing thymocyte migration: combined role of chemokines and extracellular matrix J. Leukoc. Biol., June 1, 2004; 75(6): 951 - 961. [Abstract] [Full Text] [PDF]
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K. K. Hoyer, M. Pang, D. Gui, I. P. Shintaku, I. Kuwabara, F.-T. Liu, J. W. Said, L. G. Baum, and M. A. Teitell An Anti-Apoptotic Role for Galectin-3 in Diffuse Large B-Cell Lymphomas Am. J. Pathol., March 1, 2004; 164(3): 893 - 902. [Abstract] [Full Text] [PDF]
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 T. Fukumori, Y. Takenaka, T. Yoshii, H.-R. C. Kim, V. Hogan, H. Inohara, S. Kagawa, and A. Raz CD29 and CD7 Mediate Galectin-3-Induced Type II T-Cell Apoptosis Cancer Res., December 1, 2003; 63(23): 8302 - 8311. [Abstract] [Full Text] [PDF]
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N. Nishi, H. Shoji, M. Seki, A. Itoh, H. Miyanaka, K. Yuube, M. Hirashima, and T. Nakamura Galectin-8 modulates neutrophil function via interaction with integrin {alpha}M Glycobiology, November 1, 2003; 13(11): 755 - 763. [Abstract] [Full Text] [PDF]
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K. Vandal, P. Rouleau, A. Boivin, C. Ryckman, M. Talbot, and P. A. Tessier Blockade of S100A8 and S100A9 Suppresses Neutrophil Migration in Response to Lipopolysaccharide J. Immunol., September 1, 2003; 171(5): 2602 - 2609. [Abstract] [Full Text] [PDF]
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M. J. Abedin, Y. Kashio, M. Seki, K. Nakamura, and M. Hirashima Potential roles of galectins in myeloid differentiation into three different lineages J. Leukoc. Biol., May 1, 2003; 73(5): 650 - 656. [Abstract] [Full Text] [PDF]
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Y. Kashio, K. Nakamura, M. J. Abedin, M. Seki, N. Nishi, N. Yoshida, T. Nakamura, and M. Hirashima Galectin-9 Induces Apoptosis Through the Calcium-Calpain-Caspase-1 Pathway J. Immunol., April 1, 2003; 170(7): 3631 - 3636. [Abstract] [Full Text] [PDF]
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S. G. Correa, C. E. Sotomayor, M. P. Aoki, C. A. Maldonado, and G. A. Rabinovich Opposite effects of galectin-1 on alternative metabolic pathways of L-arginine in resident, inflammatory, and activated macrophages Glycobiology, February 1, 2003; 13(2): 119 - 128. [Abstract] [Full Text] [PDF]
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 T. K. Mandal and C. Mukhopadhyay Binding free energy calculations of galectin-3-ligand interactions Protein Eng. Des. Sel., December 1, 2002; 15(12): 979 - 986. [Abstract] [Full Text] [PDF]
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G. A. Rabinovich, N. Rubinstein, and L. Fainboim Unlocking the secrets of galectins: a challenge at the frontier of glyco-immunology J. Leukoc. Biol., May 1, 2002; 71(5): 741 - 752. [Abstract] [Full Text] [PDF]
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J. L. Dunphy, G. J. Barcham, R. J. Bischof, A. R. Young, A. Nash, and E. N. T. Meeusen Isolation and Characterization of a Novel Eosinophil-specific Galectin Released into the Lungs in Response to Allergen Challenge J. Biol. Chem., April 19, 2002; 277(17): 14916 - 14924. [Abstract] [Full Text] [PDF]
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S. Sato, N. Ouellet, I. Pelletier, M. Simard, A. Rancourt, and M. G. Bergeron Role of Galectin-3 as an Adhesion Molecule for Neutrophil Extravasation During Streptococcal Pneumonia J. Immunol., February 15, 2002; 168(4): 1813 - 1822. [Abstract] [Full Text] [PDF]
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J. Mengwasser, F.-T. Liu, and J. P. Sleeman Galectin-3 is strongly up-regulated in nonapoptosing mammary epithelial cells during rat mammary gland involution Glycobiology, February 1, 2002; 12(2): 129 - 134. [Abstract] [Full Text] [PDF]
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 J. Chillaron, R. Roca, A. Valencia, A. Zorzano, and M. Palacin Heteromeric amino acid transporters: biochemistry, genetics, and physiology Am J Physiol Renal Physiol, December 1, 2001; 281(6): F995 - F1018. [Abstract] [Full Text] [PDF]
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R.-Y. Yang, D. K. Hsu, L. Yu, J. Ni, and F.-T. Liu Cell Cycle Regulation by Galectin-12, a New Member of the Galectin Superfamily J. Biol. Chem., June 1, 2001; 276(23): 20252 - 20260. [Abstract] [Full Text] [PDF]
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| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
) mAb, purified
monocytes were added to the upper chambers, and the migration assay was
performed as described in Materials and Methods. Data
are the mean ± SD of three individual experiments.
, monocyte migration when equal
concentrations of galectin-3 were added to both chambers.
), or
10 mM (
) lactose (A) or sucrose (B)
and placed in the lower chambers. Purified monocytes were added to the
upper chambers, and a standard migration assay was then performed. Data
are the mean ± SD of four individual experiments.
