Pulmonary and Critical Care Section, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520
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ABSTRACT |
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The chemokine regulated on activation normal T
cells expressed and secreted (RANTES) has been implicated in eosinophil
chemotaxis in asthma and allergic diseases, which are thought to be T
helper (Th) type 2-dominated diseases. However, adoptive
transfer of Th1 cells in mice upregulates RANTES gene expression in the
lung, and increased RANTES expression has been documented in several Th1 cell-dominated conditions that are associated with neutrophilia. The in vivo role of RANTES in the pathogenesis of disease processes is
not well understood. To determine the effect of RANTES expression alone
in vivo, we generated transgenic mice that overexpress RANTES specifically in the lung in an inducible fashion. The airways of the
transgenic mice overexpressing RANTES displayed a significant increase
in neutrophil infiltration compared with that in control mice. The
increased airway neutrophilia was also evident when the transgenic mice
were tested in a murine model of allergic airway inflammation. RANTES
expression also induced expression of the chemokine genes macrophage
inflammatory protein-2, 10-kDa interferon--inducible protein, and
monocyte chemoattractant protein-1 in the lungs of the transgenic mice.
Our studies highlight a hitherto unappreciated role for RANTES in
neutrophil trafficking during inflammation. Thus increased RANTES
expression, as observed during respiratory viral infections, may play
an important role in the associated neutrophilia and exacerbations of asthma.
chemokine; transgenic mice; lung inflammation; regulated on activation normal T cells expressed and secreted
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INTRODUCTION |
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REGULATED ON ACTIVATION NORMAL T CELLS expressed and secreted (RANTES), a C-C chemokine, was initially shown to be chemotactic for T cells and monocytes (43) but has subsequently been shown to be a potent eosinophil chemoattractant (26, 30, 31, 33, 40). RANTES can be expressed by a variety of cell types including CD8+ T cells (34), endothelial cells (32), and lung epithelial cells (27, 53). RANTES mRNA has been detected in skin biopsies obtained from atopic subjects 6 h after antigen challenge (56). In other studies, an upregulation of RANTES message was observed in the airways of asthmatic patients (51), and increased levels of RANTES have been detected in the nasal aspirates of children with viral exacerbation of asthma, suggesting an important role for RANTES in this process (41, 50).
Many studies (26, 30, 31, 33, 40) have suggested an association between RANTES expression and eosinophilia in asthma and allergic diseases, which are T helper (Th) type 2 cell-dominated diseases. However, RANTES expression has also been reported in several Th1 cell-dominated inflammatory conditions that are accompanied by neutrophilia (13, 23, 38). In mice, adoptive transfer of Th2 cells caused eosinophil influx in the lungs of mice but did not cause increased RANTES expression in the lung (18; Cohn L, Yang L, and Ray P, unpublished observations). In contrast, adoptive transfer of Th1 cells resulted in upregulation of RANTES gene expression (18; Cohn et al., unpublished observations) and increased neutrophils in the lung (10). Reciprocally, a recent report indicated that RANTES may be a chemoattractant for Th1 cells (46). Thus although many studies suggest an association between RANTES and eosinophil chemotaxis in asthma and allergy, diseases orchestrated by Th2 cells, an increase in RANTES expression during Th1 cell transfer in experimental models of inflammation and in conditions of Th1 cell-induced inflammation suggests that RANTES may have an additional important function(s) in vivo.
Because multiple chemokines with overlapping kinetics and function are induced during allergic inflammation, it is difficult to ascertain the specific role of each in leukocyte chemotaxis. To study the effector function of human RANTES (hRANTES) in vivo, we generated transgenic mice that overexpress hRANTES in an inducible fashion in the lung (39). Our studies show that hRANTES overexpression alone promotes neutrophilia in the lung. An increase in lung neutrophila was also a prominent feature of airway inflammation in the context of antigen-induced inflammation in the RANTES-overexpressing mice.
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EXPERIMENTAL PROCEDURES |
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Generation of transgenic mice. To generate transgenic mice that express hRANTES specifically in the lung in an inducible manner, two DNA constructs were used. To make the Clara cell 10-kDa protein (CC10) promoter/reverse tetracycline transactivator (nrtTA) construct (39), a partial 2.1-kb human growth hormone (hGH) gene sequence beginning at the BamH I site in the first exon to the Not I site after the polyadenylation signal isolated from the plasmid p1017 (9) was cloned into pBlueScript II KS+ (Stratagene) between the same sites to provide intronic and polyadenylation sequences. Next, a 3.0-kb Hind III fragment containing the CC10 promoter was isolated (48) and inserted at the Hind III site in the construct in the appropriate orientation. The nrtTA fragment was isolated by digesting pUHD172-neo (19) with EcoR I followed by filling in with Klenow polymerase and digesting with BamH I. This nrtTA fragment was inserted between the EcoR V and BamH I sites in the construct, yielding the final CC10 promoter/nrtTA construct. This transgene construct can express a nuclear localization signal containing a tetracycline-inducible transactivator in a lung-specific fashion. To make the plasmid construct tetracycline operator (TetOp)/hRANTES, the pBlueScript II KS+ plasmid containing the hGH gene (described above) was used as the starting plasmid. An Xho I-Cla I fragment containing the TetOp/human cytomegalovirus minimal promoter (39) was inserted between the same sites in the pBlueScript II KS+ plasmid containing the hGH gene. Next, an Xba I-Hind III fragment containing the full-length hRANTES cDNA, including its own stop codon, was inserted between the same sites in the above plasmid to generate the TetOp/hRANTES construct. The constructs were confirmed by sequencing. Transgenic mice were produced by coinjection of the two constructs into fertilized eggs from C57BL/6-SJL F2 mice.
The transgenic founder mice were identified by Southern blot analysis. The probes used were a Hind III-BamH I fragment containing the nrtTA sequence from the CC10 promoter/nrtTA construct and a BamH I fragment containing the hRANTES region from the TetOp/hRANTES construct. These founder mice were crossed with either BALB/c or C57BL/6 mice, and the transgenic mice were identified by PCR. The upper primer for detecting nrtTA was 5'-GTCGCTAAAGAAAGGGAAACAA-3', and the lower primer was 5'-TTCCAAGGGCATCGGTAAACATCTG-3'. One of the primers for detecting the hRANTES transgene (upper primer) was derived from the hRANTES cDNA and the sequence was 5'-CGGCACGCCTCGCTGTCATCCT-3', whereas the lower primer was derived from the hGH sequences and its sequence was 5'-GGGCTTAGATGGCGATACTCAC-3'. Five transgenic lines that were positive for both transgenes were obtained.Induction of hRANTES expression in transgenic mice. All animals were 8-10 wk old and were maintained on normal water until used. For the induction of hRANTES expression, transgenic mice and nontransgenic littermates were given water containing doxycycline (Dox) hydrochloride (1 mg/ml Dox in 5% sucrose) in aluminum-wrapped bottles for 4-9 days or throughout the course of the experiment in the ovalbumin (Ova) antigen model. The water was changed every 3 days. Transgenic mice that did not receive Dox were supplied with sucrose (5%) water.
Northern blotting and RNase protection assay. Total cellular RNA was obtained from different tissues of transgenic mice by homogenization in TRIzol reagent (GIBCO BRL, Life Technologies, Grand Island, NY) and purification (55). Fifteen micrograms of RNA were analyzed from each tissue. Northern blotting was carried out with QuikHyb (Stratagene). The probe was made with the random labeling kit (Boehringer Mannheim, Indianapolis, IN), and the template was a BamH I fragment containing the hRANTES cDNA isolated from the TetOp/hRANTES construct.
RNA isolated from lungs was analyzed for chemokine or cytokine gene expression by RNase protection assay (RPA) with the RPA III kit (Ambion) in accordance with the manufacturer's instructions (55). The probes were made with the murine mCK-5 set for chemokine expression and the murine mCK-1 set for cytokine expression (both from PharMingen, San Diego, CA). Ten micrograms of RNA were used in each case.Bronchoalveolar lavage and lung histology.
Bronchoalveolar lavage (BAL) was performed as previously described
(55). Briefly, lungs were lavaged with 1 ml of Dulbecco's phosphate-buffered saline (DPBS), cells were pelleted, and the supernatant was divided into aliquots and stored at 70°C. The pelleted cells were resuspended in DPBS, counted, cytospun, and stained
with Diff-Quik (Baxter HealthCare, Miami, FL). Cell differentials were
enumerated based on morphology and staining profiles. For histology,
the lungs were fixed in Streck tissue fixative (Streck Laboratories)
after lavage and stained with hematoxylin and eosin.
ELISA. The BAL fluids were analyzed for RANTES protein levels by ELISA (Endogen).
Antigen sensitization and challenge of mice.
Mice were sensitized and challenged as previously described
(55). Briefly, mice were sensitized by intraperitoneal
injection of 10 µg of Ova (Sigma, St. Louis, MO) and 1 mg of alum
(Resorptar, Intergen, New York, NY) on days 0 and
5. The transgenic mice were given either 5% sucrose water
without Dox (Dox) or 5% sucrose water with 1 mg/ml of Dox (+Dox)
24 h before the first intraperitoneal injection. On
day 12, mice were aerosol challenged by nebulization with
1% Ova (0.01% Tween 20 and DPBS) twice, for 1 h each time, with
an interval period of 4 h between the challenges. Mice were anesthetized and analyzed 24 h after the second aerosol challenge.
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RESULTS |
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Inducible lung-specific expression of RANTES.
To determine the specific role of RANTES in leukocyte chemotaxis during
lung inflammation, we generated transgenic mice that overexpress RANTES
in the lung in an inducible fashion with the use of a lung-specific
tetracycline-inducible system previously developed in our laboratory
(39) (Fig. 1A).
In this system, the nrtTA driven by the lung-specific CC10 promoter
is constitutively expressed in lung epithelial cells. When Dox
is introduced through the drinking water, it induces the binding of the
transactivator nrtTA to Tet Op/promoter sequences, resulting in
induction of expression of the linked transgene (hRANTES) from a second
construct (39). In this study, five transgenic lines were
obtained. In the presence of Dox, RANTES expression was detected only
in the lung but not in the other tissues examined (Fig. 1B).
Because of the high degree of homology between human and murine RANTES, the antibodies used in the ELISA to detect RANTES protein expression detected both endogenous and transgene expression. As shown in Fig.
2, nontransgenic mice displayed a low
level of expression (mean = 16.8 pg/ml). In the absence of Dox,
the transgenic lines 1-3 showed slightly higher levels
of RANTES expression, possibly resulting from a low level of leaky
transgene expression. Transgenic mice in line 4 showed low transgene expression in the presence and absence of Dox.
Mice in line 2 showed the highest level of Dox-induced
RANTES expression (average = 181.5 pg/ml). The basal expression of
RANTES protein in transgenic mice in line 5 was comparable
to that in their nontransgenic littermates, and there was a fourfold
increase in expression upon induction with Dox (average = 69.9 pg/ml; Fig. 2). Therefore, animals from both lines 2 and
5 were used for further analysis.
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Lung-specific expression of hRANTES preferentially recruits
neutrophils into the lung.
To determine the effect of RANTES overexpression on leukocyte
chemotaxis in the lung, mice were supplied with Dox-containing water,
the lungs of the transgenic mice and their nontransgenic littermates
were examined histologically, and cell recruitment in the airways was
assessed by BAL. As expected, in nontransgenic mice, the majority of
BAL fluid cells were macrophages (Fig.
3A). When RANTES expression
was induced in the transgenic mice, a significant increase in the
number of neutrophils was observed in the BAL fluid (10-30% of
total cells; Fig. 3, B and D). The
increase in neutrophils was observed within 24 h of the
induction of transgene expression and could still be observed after 12 days, the extent to which it was studied (Table
1). Consistent with the described role of
RANTES in T-cell chemotaxis (44), the lymphocyte
population in the BAL fluid of the transgenic mice was also found to
increase, from ~5% after 24 h to ~20% of total cells after 7 days, and was maintained at 7-10% of the total population at 12 days after induction of RANTES expression (Fig. 3E).
Surprisingly, no appreciable eosinophil influx could be observed in the
airways of the RANTES-overexpressing mice at any of the time points
tested. Although neutrophils were increased in the airways of the mice,
the lung parenchyma did not show increased neutrophil numbers or any
inflammation (Fig. 3, C and D); therefore, the
chemotactic gradient setup in these mice favored neutrophil migration
into the airways. Similarly, in a study by Gunn et al.
(21), monocyte chemoattractant protein (MCP)-1
overexpression in the lung did not cause any lung inflammation but
resulted in increased monocyte and lymphocyte infiltration into the
airways. Curiously, the increased neutrophil chemotaxis induced by
RANTES was much more prominent in transgenic mice on a BALB/c
background than in those on a C57BL/6 background (data not shown). We
therefore estimated the peripheral blood neutrophil and eosinophil
numbers in the BALB/c and C57BL/6 mice. As shown in Table
2, neutrophils account for ~25% of the
circulating leukocytes in BALB/c mice but only 10% of the total cells
in C57BL/6 mice. Thus under basal conditions, BALB/c mice have more
circulating neutrophils, which may explain the difference in neutrophil
recruitment seen in the two strains of mice.
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Lung-specific expression of hRANTES can activate the expression of
macrophage inflammatory protein-2, 10-kDa interferon--inducible
protein, and MCP-1 in the lung.
Because an association between RANTES and neutrophils is not well
described in the literature, we were interested in determining whether
RANTES overexpression caused the induction of other cytokines and/or
chemokines that might explain the increased neutrophil recruitment to
the lungs of the mice. To determine this, RPAs were performed. As
observed by us (55) and by others (29), a low
level of RANTES is constitutively expressed in the lungs of mice. As
shown in Fig. 4, the message for
macrophage inflammatory protein (MIP)-2, a potent neutrophil
chemoattractant, was detectable in the lungs of the transgenic mice. In
addition to MIP-2 mRNA, a prominent increase in expression of 10-kDa
interferon-
-inducible protein (IP-10) and MCP-1 mRNAs was also
evident within 24 h of Dox induction in the lungs of the
transgenic mice (Fig. 4, A and B). The expression
of eotaxin, lymphotactin, MIP-1
, MIP-1
, or T-cell activation gene
mRNA was not increased by RANTES overexpression (Fig. 4). RPA
did not detect increased expression of any of the cytokines tested,
including interleukin (IL)-4, IL-5, IL-10, IL-13, IL-9, IL-15, IL-2,
IL-6, and interferon-
(data not shown).
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Increased neutrophilia in RANTES-overexpressing mice in a murine
model of airway inflammation.
To determine the effect of RANTES overexpression on leukocyte
chemotaxis during inflammation, transgenic mice and nontransgenic littermate controls were treated with Dox, sensitized with Ova, and
then challenged with Ova by inhalation after 12 days. Twenty-four hours
after challenge with Ova, mice were anesthetized, BAL fluid was
recovered from the animals, and the cell differential in the BAL fluid
was determined. The total cell count in the BAL fluid recovered from
the transgenic mice on antigen challenge was three- to fourfold greater
than that in the BAL fluid derived from the nontransgenic mice (Fig.
5). After antigen provocation, although neutrophils comprised only 1-2% of the total cells present in the
airways of the nontransgenic littermates, they accounted for ~30-40% of the total cells in the airways of the
RANTES-overexpressing mice (Fig. 5). This effect was seen in both
transgenic lines 2 and 5 and was also observed in
mice on both C57BL/6 and BALB/c backgrounds. However, it was more
easily distinguishable in C57BL/6 mice, mice that typically show lower
neutrophil numbers in the lung than in BALB/c mice on antigen
provocation in this model of allergic inflammation. Also, compared with
Ova-challenged nontransgenic littermate controls, the transgenic mice
that received Dox water also had slightly more eosinophils,
lymphocytes, and macrophages in their airways. Our experiments suggest
that although RANTES alone cannot promote neutrophilia in naive C57BL/6
mice (which have lower basal neutrophil counts compared with BALB/c
mice), when neutrophilia is induced in the context of allergic
inflammation, RANTES is a strong stimulus for the recruitment of these
neutrophils into the lung. It will be interesting to determine whether
the difference in lung neutrophilia seen in the BALB/c and C57BL/6 mice
in allergic inflammation is at least partly because of differential RANTES production in the mice.
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DISCUSSION |
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Studies of chemokine function in vivo with the use of constitutive overexpression transgenic systems have been hampered by problems associated with receptor downmodulation that results from chronic stimulation by the ligand. In the present studies, we used an inducible lung-specific transgenic system to study the function of the chemokine RANTES in vivo. This system can be adapted to many cell types, including lymphocytes, with appropriate cell-specific promoters. RANTES, a member of the C-C chemokine family, is best known as a chemotactic factor for eosinophils (26, 30, 31, 33, 40) and memory T cells (43). However, recent studies (1, 3) suggest that RANTES may have biological functions that are distinct from its chemotactic effects on eosinophils. Our results show that RANTES overexpression causes increased neutrophil infiltration into the airways of mice in both the presence and absence of antigen-induced airway inflammation. Furthermore, the RANTES-overexpressing mice displayed increased expression of MIP-2, IP-10, and MCP-1 mRNA in their lungs.
Previously, RANTES was implicated in eosinophil chemotaxis based on blocking studies with antibodies (28, 54) as well as on studies that used the RANTES receptor antagonist Met-RANTES (15, 18). A surprising finding in our study is the presence of lung neutrophilia instead of eosinophilia in the RANTES-expressing mice. The principal RANTES-binding receptors are CCR1 (17, 35, 36), CCR3 (12, 36), and CCR5 (11, 37, 42). A recent study (20) highlighted an important role for CCR3 in the recruitment of eosinophils in the lung and airways of mice during eosinophilic inflammation. Although human and murine RANTES have a high degree of homology, in vitro studies (20, 36) have shown that, compared with murine CCR3, hRANTES has a low affinity for murine CCR3, the principal chemokine receptor expressed by eosinophils. Because we have expressed hRANTES in the transgenic mice, this may explain the lack of a substantial increase in eosinophils in the airways of the transgenic mice during antigen-induced inflammation. Thus our studies confirm that hRANTES is unable to cause increased chemotaxis of murine eosinophils, probably because of a low affinity for CCR3. On the other hand, because increased lymphocyte recruitment was observed in these mice, hRANTES may not discriminate between human or murine CCR1 and CCR5 in vivo. Indeed, human and murine RANTES have been shown to have similar affinities for CCR1 (36).
The increased lung neutrophilia observed in our study may be explained
as follows. First, the effect on neutrophil chemotaxis may not be
direct but may be mediated by other chemokines or mediators because
chemokines have been shown to regulate the expression of other
chemokines (18). Of the chemokines tested, RANTES
increased the expression of the chemokine genes MIP-2, IP-10, and MCP-1 in vivo. Of these chemokines, MIP-2 is a potent neutrophil
chemoattractant (2, 14, 16). In a model of bacterial
meningitis (14), a condition that involves excessive
neutrophilia in the meninges, coordinate expression of MIP-2, RANTES,
MCP-1, and MIP-1 was noted, with the kinetics of expression
paralleling disease severity. Also, in mice in which Th1 cells were
transferred or in mice infected with influenza virus, the expression of
RANTES, MCP-1, and IP-10 mRNAs was increased (29); the
effect on MIP-2 expression was not analyzed in this study.
Interestingly, both IP-10 and MCP-1 have been implicated in neutrophil
trafficking. Subcutaneous injection of IP-10 in BALB/c mice was
associated with infiltration of both mononuclear cells and neutrophils
in the subcutaneous tissue within 4 h of injection
(49). MCP-1 was shown to behave as an efficient neutrophil
chemoattractant in mice in the context of chronic inflammation (25). Similar to the potent effect of low doses of MCP-1
on neutrophils in the presence of inflammation, the local expression of
multiple chemokines, RANTES, IP-10, and MCP-1 may have the ability to
set up an efficient chemotactic gradient for neutrophils as observed in
the RANTES transgenic mice. When viewed in combination, in a Th1
cell-dominated inflammation, RANTES may be the primary chemokine
activated by Th1 cells, which, in turn, sets up a network of other
chemokines such as MIP-2, IP-10, and MCP-1. To the best of our
knowledge, this is the first report that shows that RANTES can induce
the expression of other chemokine genes. It will be interesting to
determine which chemokine receptor is responsible for the increased
neutrophil chemotaxis by breeding the RANTES transgenic mice to
specific chemokine receptor-deficient mice.
Second, several biological effects of RANTES have been suggested to occur in an aggregation-dependent manner. For example, in a recently described study (1), aggregated but not disaggregated hRANTES was shown to activate human neutrophils with a substantial increase in CD11b expression. Thus aggregation of RANTES in vivo may also be responsible for its neutrophil chemoattractant properties. Because resting neutrophils were shown to express CCR1 (6), it is possible that this basal level of CCR1 expression is sufficient for responsiveness to aggregated RANTES.
In the lung, Th1 cell-dominated inflammation induces neutrophilia,
whereas Th2 cell-dominated inflammation promotes eosinophilia, as has
been clearly demonstrated by adoptive transfer experiments (10). RANTES expression has been demonstrated in several
Th1 cell-dominated conditions such as rheumatoid arthritis
(38), delayed-type hypersensitivity (13), and
sarcoidosis (23). In other studies, Schrum et al.
(45) showed a strong expression of RANTES in a Th1
cell-type response to a gram-negative bacterium but a low expression in
the Th2-type response to a nematode infection. Recently, in a
murine model of rheumatoid arthritis (3), neutralization of RANTES, but not of other chemokines, greatly attenuated symptoms of
the disease. In adoptive transfer experiments, Th1, but not Th2, cells
were found to upregulate RANTES mRNA expression in the lung,
suggesting an association between RANTES and Th1 cell-mediated effects in the lung (18; Cohn et al., unpublished observations). Also,
RANTES causes chemotaxis of Th1 cells (46) and,
reciprocally, interferon- was shown to increase RANTES
expression, whereas IL-4 and IL-13 decreased expression (5, 24,
32). Although several studies show a close association between
RANTES expression and a Th1 cell-type response, the
specific role of RANTES in these conditions is unclear.
Our studies suggest that directly or indirectly RANTES may be involved
in the neutrophilia associated with Th1 cell-dominated diseases. Thus
increased RANTES expression, as observed during respiratory viral
infections, may play an important role in the associated neutrophilia
and exacerbation of asthma (4, 7, 8, 22, 41, 47, 52). The
detailed mechanism of RANTES-induced neutrophilia will be carried out
in the next phase of this study. With antibody neutralization
experiments and appropriate receptor-deficient mice, the direct or
indirect effect of RANTES on neutrophil biology will be addressed. The use of receptor knockout mice will be particularly useful in
determining the differential role of RANTES in vivo.
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ACKNOWLEDGEMENTS |
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We thank Drs. M. Gossen and H. Bujard for the gift of the plasmid pUHD172-neo containing the reverse tetracycline transactivator, Drs. J. Whitsett and B. Stripp for the gift of the plasmid pCC10CAT-2300 containing the Clara cell 10-kDa protein (CC10) promoter, R. Homer for expert help with the cell differentials, and R. Swinsick for the maintenance of the mouse colonies and assistance with the mouse tail biopsies.
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FOOTNOTES |
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This work was supported by National Heart, Lung, and Blood Institute Grants HL-60207 (to P. Ray) and HL-60995 (to A. Ray and P. Ray).
Address for reprint requests and other correspondence: P. Ray, Dept. of Internal Medicine, Pulmonary and Critical Care Section, Yale Univ. School of Medicine, 333 Cedar St., LCI 105, New Haven, CT 06520 (E-mail: Prabir.Ray{at}yale.edu).
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. Section 1734 solely to indicate this fact.
Received 3 March 2000; accepted in final form 19 April 2000.
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REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Appay, V,
Brown A,
Cribbes S,
Randle E,
and
Czaplewski LG.
Aggregation of RANTES is responsible for its inflammatory properties. Characterization of nonaggregating, noninflammatory RANTES mutants.
J Biol Chem
274:
27505-27512,
1999
2.
Appelberg, R.
Macrophage inflammatory proteins MIP-1 and MIP-2 are involved in T-cell-mediated neutrophil recruitment.
J Leukoc Biol
52:
303-306,
1992[Abstract].
3.
Barnes, DA,
Tse J,
Kaufhold M,
Owen M,
Hesselgesser J,
Strieter R,
Horuk R,
and
Perez HD.
Polyclonal antibody directed against human RANTES ameliorates disease in the Lewis rat adjuvant-induced arthritis model.
J Clin Invest
101:
2910-2919,
1998
4.
Becker, S,
Reed W,
Henderson FW,
and
Noah TL.
RSV infection of human airway epithelial cells causes production of the -chemokine RANTES.
Am J Physiol Lung Cell Mol Physiol
272:
L512-L520,
1997
5.
Berkman, N,
Krishnan VL,
Gilbey T,
Newton R,
O'Connor B,
Barnes PJ,
and
Chung KF.
Expression of RANTES mRNA and protein in airways of patients with mild asthma.
Am J Respir Crit Care Med
154:
1804-1811,
1996[Abstract].
6.
Bonecchi, R,
Polentarutti N,
Luini W,
Borsatti A,
Bernasconi S,
Locati M,
Power C,
Proudfoot A,
Wells TN,
Mackay C,
Mantovani A,
and
Sozzani S.
Up-regulation of CCR1 and CCR3 and induction of chemotaxis to CC chemokines by IFN- in human neutrophils.
J Immunol
162:
474-479,
1999
7.
Busse, WW,
Vrtis RF,
Steiner R,
and
Dick EC.
In vitro incubation with influenza virus primes human polymorphonuclear leukocyte generation of superoxide.
Am J Respir Cell Mol Biol
4:
347-354,
1991[ISI][Medline].
8.
Calhoun, WJ,
Dick EC,
Schwartz LB,
and
Busse WW.
A common cold virus, rhinovirus 16, potentiates airway inflammation after segmental antigen bronchoprovocation in allergic subjects.
J Clin Invest
94:
2200-2208,
1994[ISI][Medline].
9.
Chaffin, KE,
Beals CR,
Wilkie TM,
Forbush KA,
Simon MI,
and
Perlmutter RM.
Dissection of thymocyte signaling pathways by in vivo expression of pertussis toxin ADP-ribosyltransferase.
EMBO J
9:
3821-3829,
1990[Abstract].
10.
Cohn, L,
Homer RJ,
Marinov A,
Rankin J,
and
Bottomly K.
Induction of airway mucus production by T helper 2 (Th2) cells: a critical role for interleukin 4 in cell recruitment but not mucus production.
J Exp Med
186:
1737-1747,
1997
11.
Combadiere, C,
Ahuja SK,
Tiffany HL,
and
Murphy PM.
Cloning and functional expression of CC CKR5, a human monocyte CC chemokine receptor selective for MIP-1(), MIP-1(
), and RANTES.
J Leukoc Biol
60:
147-152,
1996[Abstract].
12.
Daugherty, BL,
Siciliano SJ,
DeMartino JA,
Malkowitz L,
Sirotina A,
and
Springer MS.
Cloning, expression, and characterization of the human eosinophil eotaxin receptor.
J Exp Med
183:
2349-2354,
1996[Abstract].
13.
Devergne, O,
Marfaing-Koka A,
Schall TJ,
Leger-Ravet MB,
Sadick M,
Peuchmaur M,
Crevon MC,
Kim KJ,
Schall TT,
Kim T,
Galanaud P,
and
Emilie D.
Production of the RANTES chemokine in delayed-type hypersensitivity reactions: involvement of macrophages and endothelial cells.
J Exp Med
179:
1689-1694,
1994[Abstract].
14.
Diab, A,
Abdalla H,
Li HL,
Shi FD,
Zhu J,
Hojberg B,
Lindquist L,
Wretlind B,
Bakhiet M,
and
Link H.
Neutralization of macrophage inflammatory protein 2 (MIP-2) and MIP-1 attenuates neutrophil recruitment in the central nervous system during experimental bacterial meningitis.
Infect Immun
67:
2590-2601,
1999
15.
Elsner, J,
Petering H,
Kimmig D,
Wells TN,
Proudfoot AE,
and
Kapp A.
The C-C chemokine receptor antagonist met-RANTES inhibits eosinophil effector functions.
Int Arch Allergy Immunol
118:
462-465,
1999[ISI][Medline].
16.
Feng, L,
Xia Y,
Yoshimura T,
and
Wilson CB.
Modulation of neutrophil influx in glomerulonephritis in the rat with anti-macrophage inflammatory protein-2 (MIP-2) antibody.
J Clin Invest
95:
1009-1017,
1995[ISI][Medline].
17.
Gao, JL,
Kuhns DB,
Tiffany HL,
McDermott D,
Li X,
Francke U,
and
Murphy PM.
Structure and functional expression of the human macrophage inflammatory protein 1/RANTES receptor.
J Exp Med
177:
1421-1427,
1993[Abstract].
18.
Gonzalo, JA,
Lloyd CM,
Wen D,
Albar JP,
Wells TN,
Proudfoot A,
Martinez AC,
Dorf M,
Bjerke T,
Coyle AJ,
and
Gutierrez-Ramos JC.
The coordinated action of CC chemokines in the lung orchestrates allergic inflammation and airway hyperresponsiveness.
J Exp Med
188:
157-167,
1998
19.
Gossen, M,
Freundlieb S,
Bender G,
Muller G,
Hillen W,
and
Bujard H.
Transcriptional activation by tetracyclines in mammalian cells.
Science
268:
1766-1769,
1995[ISI][Medline].
20.
Grimaldi, JC,
Yu NX,
Grunig G,
Seymour BW,
Cottrez F,
Robinson DS,
Hosken N,
Ferlin WG,
Wu X,
Soto H,
O'Garra A,
Howard MC,
and
Coffman RL.
Depletion of eosinophils in mice through the use of antibodies specific for C-C chemokine receptor 3 (CCR3).
J Leukoc Biol
65:
846-853,
1999[Abstract].
21.
Gunn, MD,
Nelken NA,
Liao X,
and
Williams LT.
Monocyte chemoattractant protein-1 is sufficient for the chemotaxis of monocytes and lymphocytes in transgenic mice but requires an additional stimulus for inflammatory activation.
J Immunol
158:
376-383,
1997[Abstract].
22.
Holgate, ST,
Bodey KS,
Janezic A,
Frew AJ,
Kaplan AP,
and
Teran LM.
Release of RANTES, MIP-1, and MCP-1 into asthmatic airways following endobronchial allergen challenge.
Am J Respir Crit Care Med
156:
1377-1383,
1997
23.
Iida, K,
Kadota J,
Kawakami K,
Matsubara Y,
Shirai R,
and
Kohno S.
Analysis of T cell subsets and -chemokines in patients with pulmonary sarcoidosis.
Thorax
52:
431-437,
1997[Abstract].
24.
John, M,
Hirst SJ,
Jose PJ,
Robichaud A,
Berkman N,
Witt C,
Twort CH,
Barnes PJ,
and
Chung KF.
Human airway smooth muscle cells express and release RANTES in response to T helper 1 cytokines: regulation by T helper 2 cytokines and corticosteroids.
J Immunol
158:
1841-1847,
1997[Abstract].
25.
Johnston, B,
Burns AR,
Suematsu M,
Issekutz TB,
Woodman RC,
and
Kubes P.
Chronic inflammation upregulates chemokine receptors and induces neutrophil migration to monocyte chemoattractant protein-1.
J Clin Invest
103:
1269-1276,
1999
26.
Kameyoshi, Y,
Dorschner A,
Mallet AI,
Christophers E,
and
Schroder JM.
Cytokine RANTES released by thrombin-stimulated platelets is a potent attractant for human eosinophils.
J Exp Med
176:
587-592,
1992[Abstract].
27.
Kwon, OJ,
Jose PJ,
Robbins RA,
Schall TJ,
Williams TJ,
and
Barnes PJ.
Glucocorticoid inhibition of RANTES expression in human lung epithelial cells.
Am J Respir Cell Mol Biol
12:
488-496,
1995[Abstract].
28.
Lampinen, M,
Rak S,
and
Venge P.
The role of interleukin-5, interleukin-8 and RANTES in the chemotactic attraction of eosinophils to the allergic lung.
Clin Exp Allergy
29:
314-322,
1999[ISI][Medline].
29.
Li, L,
Xia Y,
Nguyen A,
Feng L,
and
Lo D.
Th2-induced eotaxin expression and eosinophilia coexist with Th1 responses at the effector stage of lung inflammation.
J Immunol
161:
3128-3135,
1998
30.
Lukacs, NW,
Standiford TJ,
Chensue SW,
Kunkel RG,
Strieter RM,
and
Kunkel SL.
C-C chemokine-induced eosinophil chemotaxis during allergic airway inflammation.
J Leukoc Biol
60:
573-578,
1996[Abstract].
31.
Lukacs, NW,
Strieter RM,
Warmington K,
Lincoln P,
Chensue SW,
and
Kunkel SL.
Differential recruitment of leukocyte populations and alteration of airway hyperreactivity by C-C family chemokines in allergic airway inflammation.
J Immunol
158:
4398-4404,
1997[Abstract].
32.
Marfaing-Koka, A,
Devergne O,
Gorgone G,
Portier A,
Schall TJ,
Galanaud P,
and
Emilie D.
Regulation of the production of the RANTES chemokine by endothelial cells. Synergistic induction by IFN- plus TNF-
and inhibition by IL-4 and IL-13.
J Immunol
154:
1870-1878,
1995
33.
Meurer, R,
Van Riper G,
Feeney W,
Cunningham P,
Hora D, Jr,
Springer MS,
MacIntyre DE,
and
Rosen H.
Formation of eosinophilic and monocytic intradermal inflammatory sites in the dog by injection of human RANTES but not human monocyte chemoattractant protein 1, human macrophage inflammatory protein 1, or human interleukin 8.
J Exp Med
178:
1913-1921,
1993[Abstract].
34.
Nelson, PJ,
Kim HT,
Manning WC,
Goralski TJ,
and
Krensky AM.
Genomic organization and transcriptional regulation of the RANTES chemokine gene.
J Immunol
151:
2601-2612,
1993
35.
Neote, K,
DiGregorio D,
Mak JY,
Horuk R,
and
Schall TJ.
Molecular cloning, functional expression, and signaling characteristics of a C-C chemokine receptor.
Cell
72:
415-425,
1993[ISI][Medline].
36.
Post, TW,
Bozic CR,
Rothenberg ME,
Luster AD,
Gerard N,
and
Gerard C.
Molecular characterization of two murine eosinophil -chemokine receptors.
J Immunol
155:
5299-5305,
1995[Abstract].
37.
Raport, CJ,
Gosling J,
Schweickart VL,
Gray PW,
and
Charo IF.
Molecular cloning and functional characterization of a novel human CC chemokine receptor (CCR5) for RANTES, MIP-1, and MIP-1
.
J Biol Chem
271:
17161-17166,
1996
38.
Rathanaswami, P,
Hachicha M,
Sadick M,
Schall TJ,
and
McColl SR.
Expression of the cytokine RANTES in human rheumatoid synovial fibroblasts. Differential regulation of RANTES and interleukin-8 genes by inflammatory cytokines.
J Biol Chem
268:
5834-5839,
1993
39.
Ray, P,
Tang W,
Wang P,
Homer R,
Kuhn C, III,
Flavell RA,
and
Elias JA.
Regulated overexpression of interleukin 11 in the lung. Use to dissociate development-dependent and -independent phenotypes.
J Clin Invest
100:
2501-2511,
1997
40.
Rot, A,
Krieger M,
Brunner T,
Bischoff SC,
Schall TJ,
and
Dahinden CA.
RANTES and macrophage inflammatory protein 1 induce the migration and activation of normal human eosinophil granulocytes.
J Exp Med
176:
1489-1495,
1992[Abstract].
41.
Saito, T,
Deskin RW,
Casola A,
Haeberle H,
Olszewska B,
Ernst PB,
Alam R,
Ogra PL,
and
Garofalo R.
Respiratory syncytial virus induces selective production of the chemokine RANTES by upper airway epithelial cells.
J Infect Dis
175:
497-504,
1997[ISI][Medline].
42.
Samson, M,
Labbe O,
Mollereau C,
Vassart G,
and
Parmentier M.
Molecular cloning and functional expression of a new human CC-chemokine receptor gene.
Biochemistry
35:
3362-3367,
1996[ISI][Medline].
43.
Schall, TJ,
Bacon K,
Toy KJ,
and
Goeddel DV.
Selective attraction of monocytes and T lymphocytes of the memory phenotype by cytokine RANTES.
Nature
347:
669-671,
1990[ISI][Medline].
44.
Schall, TJ,
Simpson NJ,
and
Mak JY.
Molecular cloning and expression of the murine RANTES cytokine: structural and functional conservation between mouse and man.
Eur J Immunol
22:
1477-1481,
1992[ISI][Medline].
45.
Schrum, S,
Probst P,
Fleischer B,
and
Zipfel PF.
Synthesis of the CC-chemokines MIP-1, MIP-1
, and RANTES is associated with a type 1 immune response.
J Immunol
157:
3598-3604,
1996[Abstract].
46.
Siveke, JT,
and
Hamann A.
T helper 1 and T helper 2 cells respond differentially to chemokines.
J Immunol
160:
550-554,
1998
47.
Stark, JM,
Godding V,
Sedgwick JB,
and
Busse WW.
Respiratory syncytial virus infection enhances neutrophil and eosinophil adhesion to cultured respiratory epithelial cells. Roles of CD18 and intercellular adhesion molecule-1.
J Immunol
156:
4774-4782,
1996
48.
Stripp, BR,
Sawaya PL,
Luse DS,
Wikenheiser KA,
Wert SE,
Huffman JA,
Lattier DL,
Singh G,
Katyal SL,
and
Whitsett JA.
cis-acting elements that confer lung epithelial cell expression of the CC10 gene.
J Biol Chem
267:
14703-14712,
1992
49.
Taub, DD,
Longo DL,
and
Murphy WJ.
Human interferon-inducible protein-10 induces mononuclear cell infiltration in mice and promotes the migration of human T lymphocytes into the peripheral tissues and human peripheral blood lymphocytes-SCID mice.
Blood
87:
1423-1431,
1996
50.
Teran, L,
Johnston SL,
and
Holgate ST.
Immunoreactive RANTES and MIP-1 are secreted into the nasal aspirates of children with virus-associated asthma (Abstract).
Am J Respir Crit Care Med
151:
A385,
1995.
51.
Teran, LM,
Noso N,
Carroll M,
Davies DE,
Holgate S,
and
Schroder JM.
Eosinophil recruitment following allergen challenge is associated with the release of the chemokine RANTES into asthmatic airways.
J Immunol
157:
1806-1812,
1996[Abstract].
52.
Teran, LM,
Seminario MC,
Shute JK,
Papi A,
Compton SJ,
Low JL,
Gleich GJ,
and
Johnston SL.
RANTES, macrophage-inhibitory protein 1, and the eosinophil product major basic protein are released into upper respiratory secretions during virus-induced asthma exacerbations in children.
J Infect Dis
179:
677-681,
1999[ISI][Medline].
53.
VanOtteren, GM,
Strieter RM,
Kunkel SL,
Paine R, III,
Greenberger MJ,
Danforth JM,
Burdick MD,
and
Standiford TJ.
Compartmentalized expression of RANTES in a murine model of endotoxemia.
J Immunol
154:
1900-1908,
1995
54.
Venge, J,
Lampinen M,
Hakansson L,
Rak S,
and
Venge P.
Identification of IL-5 and RANTES as the major eosinophil chemoattractants in the asthmatic lung.
J Allergy Clin Immunol
97:
1110-1115,
1996[ISI][Medline].
55.
Yang, L,
Cohn L,
Zhang DH,
Homer R,
Ray A,
and
Ray P.
Essential role of nuclear factor-B in the induction of eosinophilia in allergic airway inflammation.
J Exp Med
188:
1739-1750,
1998
56.
Ying, S,
Taborda-Barata L,
Meng Q,
Humbert M,
and
Kay AB.
The kinetics of allergen-induced transcription of messenger RNA for monocyte chemotactic protein-3 and RANTES in the skin of human atopic subjects: relationship to eosinophil, T cell, and macrophage recruitment.
J Exp Med
181:
2153-2159,
1995[Abstract].