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Department of Pediatrics, Case Western Reserve University, Cleveland, OH 44106
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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CF lung disease is characterized by chronic endobronchial infection and
extremely high concentrations of the proinflammatory cytokines TNF-
,
IL-1, IL-6, and especially IL-8 (2, 4, 5, 6). Together, they
result in a massive and persistent influx of neutrophils into airways
(2, 7). Multiple lines of evidence suggest that this
neutrophil influx is excessive relative to the burden of chronic
infection in these patients (2, 8, 9). Eventually, the
neutrophils and their products establish a vicious cycle of
inflammation that escapes from homeostatic control and claims the life
of the patient (10). Studies from our own (1, 2, 4) and other laboratories (3, 11) suggest that
IL-10 deficiency in the CF lung deprives that environment of a vital
"cytokine synthesis inhibitory factor" (12) and thus
plays a major role in the dysregulation of the local inflammatory
response. Animal studies have shown that treatment with exogenous IL-10
can ameliorate the excessive inflammation in models of chronic
Pseudomonas aeruginosa infection which mimic that found in
CF patients (13), and that IL-10 deficiency is associated
with increased inflammation and more severe systemic effects in these
models.
Studies of fresh bronchoalveolar lavage macrophages (BALMs) from CF patients show that they are actively producing the proinflammatory cytokines whose concentrations are highly elevated in the bronchoalveolar fluid of CF patients (2, 4). However, the Ag-presenting activity of these cells has not been well investigated. Although most CF patients have hypergammaglobulinemia, the role of BALMs or other APCs in stimulating the increased Ab production has not been studied. Because IL-10 may exert important immunodulatory effects by suppressing synthesis of proinflammatory cytokines and by altering Ag presentation and T cell responses, we postulated that the IL-10 deficiency in the CF airway might alter the immunologic function of the macrophages in that environment.
Others have previously shown that normal BALMs have poor costimulatory activity in T cell proliferation assays and that they fail to express the T cell costimulatory molecules B7-1 and B7-2 (CD80 and CD86, respectively) (14). It has also been shown that IL-10 down-regulates the expression of MHC class II (MHC-II) (15) and inhibits expression of B7 molecules by blood monocytes in response to LPS stimulation in vitro (16). Therefore, we reasoned that constitutive IL-10 production by normal bronchial epithelial cells might be responsible for the lack of B7 expression on BALMs in the healthy lung in vivo and that expression of these costimulatory molecules might be increased in CF. To test these hypotheses, we used immunostaining and flow cytometry to determine whether B7 expression on BALMs would be increased in IL-10 knockout and cystic fibrosis transmembrane conductance regulator (CFTR) knockout mice and whether the increased B7 expression in the latter would be reversed by treatment with exogenous rIL-10. The costimulatory activity of BALMs of different origins (IL-10+/+ vs IL-10-/- and CFTR+/+ vs CFTR-/-) for purified splenic T cells was also determined. The results suggest that the IL-10 secreted by normal lung epithelial cells may be an important determinant of the local immunologic milieu, which could be altered in disease states that affect the IL-10 production, like CF.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Bronchoscopy and BAL were performed on healthy individuals and clinically stable CF patients as previously described by Konstan et al. (7). BALMs were recovered by centrifugation at 250 x g for 10 min at 4°C and used for cytospin preparations. Healthy controls and CF patients gave written informed consent, and the bronchoscopy protocol was approved by the institutional review board of University Hospitals of Cleveland (Cleveland, OH).
Mice
Wild-type (+/+) and IL-10 (B10.129P2(B6)-IL10tm1Cgn) knockout (-/-) mice on the same genetic background (C57BL/10J) (17) matched for age, weight, and sex were obtained from The Jackson Laboratory (Bar Harbor, ME). Mice homozygous for the S489X (B6.129P2-Cftrtm1Unc) mutation of the CFTR gene, congenic (n = 10 generations) onto C57BL/6J background, (18) and their normal littermates, designated as CFTR-/- and CFTR+/+, respectively, were bred in our Animal Core facility. The genotype of individual animals was established by PCR amplification of tail snip genomic DNA (19, 20). All experimental animals (IL-10+/+, IL-10-/-, CFTR+/+, and CFTR-/-) were housed in isolator cages in a specific pathogen-free environment. Autoclaved rodent chow and water and liquid Peptamen diet (Clintec Nutrition, Deerfield, IL) for the CFTR-/- mice were available ad libitum. All protocols were approved by the Case Western Reserve University Institutional Animal Care and Use Committee.
Mouse BALMs
BAL was performed on anesthetized mice using six 0.5-ml aliquots of sterile normal saline as previously described (13). To obtain sufficient numbers of cells for the desired analyses, cells from three to five mice were pooled for each experiment. Aliquots of each individual animal’s BAL fluid (200 µl) were cultured on tryptic soy agar plates with 5% sheep blood to assure that the mice were not infected. Pelletted cells and BAL fluid were used for FACS and cytokine analyses. The total and differential cell count from each animal were determined with a hemacytometer and Wright-Giemsa staining of cytospin preparations, respectively. In general, BAL cells of all animals were >95% macrophages.
Immunofluorescent microscopy analysis of B7-1 costimulatory molecule
Cytospin slides made with 5 x 104
cells, each using 1% BSA in PBS, were allowed to air dry and were
fixed with 4% paraformaldehyde in PBS. Fixed cells were gently washed
with PBS, blocked for 1 h with 5% goat serum in PBS, and
incubated overnight with anti-CD80 Abs. For human specimens, we
used mouse anti-human CD80 (Ancell, Bayport, MN), followed by five
washes with 1% BSA in PBS and 1 h incubation with secondary
FITC-labeled goat anti-mouse Ab (Jackson ImmunoResearch
Laboratories, West Grove, PA). Controls with isotype-matched primary
and the same secondary Abs were used to determine background. For mouse
cells, we used hamster anti-mouse CD80 Ab directly labeled with
FITC (BD PharMingen, San Diego, CA). Isotype-matched direct
conjugates from the same vendor were used as controls. After incubation
with FITC-labeled Abs, the slides were washed five times with 1% BSA
in PBS, mounted with 9:1 glycerol-PBS, and visualized by fluorescent
microscopy. For a positive control in the mouse studies, we treated
BALMs from wild-type mice in vitro on Lab-Tek 8 plates (5 x
105 cells/well; Fisher, Pittsburgh, PA) with or
without IFN-
(100 ng/ml; R&D Systems, Minneapolis, MN) overnight.
After 24 h, the medium was removed and the adherent cells were
fixed and stained as described above.
ELISA
BAL fluid was lyophilized and then reconstituted in one-third of its original volume with PBS, and thus was concentrated 3-fold. IL-10 was then assayed using commercially available ELISA kits and recombinant standards from Endogen (Woburn, MA) in an automated reader (ThermoMax Microplate Reader, Molecular Devices, Sunnyvale, CA). The results were standardized for dilution of epithelial lining fluid using the urea method (21).
Flow cytometry analysis of B7-1 (CD80) and B7-2 (CD86) costimulatory molecules
Aliquots of 5 x 105 pooled BALMs
were first incubated with anti-mouse CD32/CD16 mAbs (2
µl/106 cells) at 4°C for 10 min to block
FcR. Cells were incubated for 30 min with biotinylated primary Abs
(hamster or rat anti-mouse CD80, CD86, or MHC II; BD PharMingen,
San Diego, CA) or with species- and isotype-matched control Abs and
then were washed three times in FACS buffer (HBSS, 10 mM of HEPES,
0.1% BSA, 0.1% NaN3) and incubated with a 1/200
dilution of streptavidin-FITC or a 1/2000 dilution of streptavidin-PE
(BD PharMingen, San Jose, CA). BALMs were washed again, resuspended in
FACS buffer, and analyzed by flow cytometry. Data for 5000 cells
falling within appropriate forward and side light scatter gates were
collected from each sample with a FACScan flow cytometer (BD
Biosciences, San Jose, CA). Nonspecific binding (background) was
determined with biotinylated irrelevant primary Ab of the same isotype
and the same conjugates. A discriminator was set to include 97% of the
cells in the background histogram as negative. Cells stained with each
specific Ab with fluorescence greater than this discriminator were
considered positive. Data were analyzed with CellQuest software. The
FACScan was standardized with fluorescent Calibrite beads to
allow direct comparison of data obtained on different days (BD
Immunocytometry Systems, San Jose, CA).
T cell costimulation activity of BALMs
The ability of BALMs from IL-10+/+ vs
IL-10-/- and CFTR+/+ vs
CFTR-/- mice to provide in vitro costimulatory
activity for proliferative responses of splenic T cells to anti-CD3
mAb (BD PharMingen) was measured by
[3H]thymidine uptake (22).
Briefly, BALMs or adherent spleen APCs (30,000/well) were cocultured
with 2 x 105 splenic lymphocytes from the
same animals that had been purified using Mouse T Cell Enrichment
Columns (R&D Systems). BALMs (
95% pure) were directly added to
96-well flat-bottom tissue culture plates (Corning Glass, Corning, NY).
Spleen APCs were isolated by plastic adherence (23). T
cells were cocultured with BALMs or spleen APCs in complete RPMI 1640
containing 5% FCS and stimulated for 72 h at 36°C in 5%
CO2 with anti-CD3 mAb (0.1 µg/1 ml; BD
PharMingen). To demonstrate the dependence of this response on B7,
identical experiments were completed in the presence of 2 µg/ml of a
15-residue peptide that blocks the B7 binding site on CD28 (Santa Cruz
Biotechnology, Santa Cruz, CA). Cells were pulsed over the last 18
h of 3-day cultures with 0.25 µCi of
[3H]thymidine and then washed with an
LKB-Wallac cell harvester, and uptake of
[3H]thymidine was measured in a LKB-Wallac beta
counter (Gaithersburg, MD).
IL-10 treatment of CFTR mice
Eight 6- to 8-wk-old CFTR-/- mice weighing 20–25 g were injected i.p. daily for 2 wk with 1 µg of recombinant mouse (rm)IL-10 in 200 µl of 20 mM Tris, 0.1 M NaCl (pH 8.0), containing 50 µg of BSA (R&D Systems). Control groups of eight CFTR-/- and eight CFTR+/+ littermate mice each were injected with the same solution without IL-10. Administration of IL-10 or vehicle was tolerated very well with no apparent side effects. After 14 days, all of the mice were sacrificed and BAL was performed. BALMs were put into two pools of cells from four animals of each experimental group and evaluated as described above. Initial aliquots of BAL fluid from each animal were cultured separately to assure there was no bacterial infection.
Statistical analysis
Statistical analysis was performed by t test; p values <0.05 were considered significant. All data in text and figures are expressed as the mean ± SD.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Immunofluorescent stained BALMs from CF patients were essentially
100% positive for expression of B7-1 (Fig. 1
A), whereas cells isolated
from healthy individuals showed only minimal staining (Fig. 1B). However, because most CF patients are chronically
infected, which may increase B7 expression, these data do not
distinguish between the CFTR defect and infection as the cause of the
increased B7 expression. Therefore, we turned to animal models
(CFTR-/- vs CFTR+/+ mice
and IL-10 knockout (IL-10-/-) vs wild-type
(IL-10+/+) mice) to determine the effects of CFTR
defects and IL-10 deficiency on B7 expression in the absence of
bacterial infection. As shown in Fig. 1C, we found positive
B7-1 staining on BALMs from uninfected IL-10-/-
mice, whereas IL-10+/+ mice had no detectable
staining (Fig. 1D). Similar results were found with BALMs
from uninfected CFTR-/- mice (Fig. 1E) vs their CFTR+/+ controls (Fig. 1F). Control slides using the isotype-matched control and/or
secondary Ab alone were completely without fluorescence (data not
shown). As a positive control, BALMs from wild-type mice were treated
in vitro overnight with 100 ng/ml of IFN-. Staining of these cells
(data not shown) for CD80 was essentially uniformly positive, similar
to the CF and IL-10 knockout mice.
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We hypothesized that BAL macrophages from the
CFTR-/- and IL-10-/-
mice might have similarly up-regulated B-7 expression because we had
previously shown that BAL fluid from CF patients was deficient in IL-10
(1, 2). The observations above suggested that
CFTR-/- and IL-10 knockout mice might have
similar microenvironments in their lungs. We next wished to determine
directly whether the lungs of CFTR-/- mice were
deficient in IL-10. Therefore, we determined the IL-10 concentrations
in uninfected CFTR+/+ and
CFTR-/- mice. Data in Fig. 2 demonstrate that
CFTR-/- mice are IL-10 deficient, with only 2
of 15 fluid samples having any IL-10 that could be detected in an ELISA
with sensitivity of 20 pg/ml. In contrast, 14 of 15 BAL fluid samples
from CFTR+/+ mice contained detectable IL-10,
giving a mean concentration of 94 ± 36 pg/ml IL-10.
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To obtain more readily quantifiable results for B7 expression, we
pooled BALM cells from three to five animals of each mouse strain and
analyzed them by flow cytometry. As shown in Fig. 3, the results are comparable with those
we observed on immunofluorescence. FACS analysis showed that 63.9% of
gated BALMs from IL-10-/- mice were positive
for B7-1 (Fig. 3A), whereas only 4.4% of cells from
IL-10+/+ control mice were positive (Fig. 3B). Similarly, 67.4% of BALMs from
CFTR-/- mice were positive for B7-1 (Fig. 3C) (n = 3 experiments of five mice
in each group), whereas cells from CFTR+/+ mice
were 
5% positive (Fig. 3D). B7-2 expression was somewhat
lower than B7-1 in all groups, but its expression on
CFTR-/- and IL-10-/-
cells was also higher than on cells from their +/+ counterparts. A
summary of data on expression of both costimulatory molecules (CD80 and
CD86) as well as MHC-II is presented in Table I. It should be noted that the data shown
in the tables may underestimate the actual percentage of B7-positive
cells, because the histograms in Fig. 3, A and C,
show relative homogeneity of the B7 staining on the BALMs, but this
overlaps with the background histograms, reducing the number derived
for apparent positive cells. Similar observations were made for MHC-II
expression (histograms not shown).
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We next wished to determine whether the increased B7 expression
correlated with increased costimulatory activity for T cells. We used
anti-CD3 as a mitogen because this stimulus had previously been
shown to require costimulation via B7 (14). First, we
compared the ability of BALMs with that of adherence purified splenic
macrophages to provide costimulation with anti-CD3 Ab, using
purified splenic T cells as the responders. BALMs from
IL-10+/+ mice had no appreciable activity in
inducing proliferation, whereas adherent spleen APCs were quite
effective (Fig. 4A). In
contrast, BALMs from IL-10-/- mice showed
increased ability to induce proliferation in the presence of
anti-CD3 (Fig. 4B, left); however, their
splenic APCs did not differ from the splenic APCs of the +/+ mice in
this assay of costimulatory activity. The use of a synthetic peptide
that inhibits the B7 binding to its receptor on T cells (CD28) shows
that, particularly with the BALMs, this costimulatory effect was B7
dependent. Next we performed similar comparisons of the costimulatory
activity of BALMs vs splenic APCs from CFTR+/+
and CFTR-/- mice. As shown in Fig. 5, BALMs from control
CFTR+/+ mice had very little costimulatory
activity (left). In contrast, consistent with their
increased B7 expression, BALMs from CFTR-/-
mice (right) induced greater proliferation by the T cells
(p > 0.05) compared with BALMs from
CFTR+/+ mice. The activity of splenic APCs from
both of these types of mice (not shown in figure) was similar to each
other and to that of IL-10+/+ and
IL-10-/- mice. Again, the costimulatory
activity of the BALMs was significantly inhibited by the peptide which
blocked the binding site for B7 on CD28 (filled bars). Probably because
the monomeric soluble peptide does not bind as well as the multivalent
interaction with native B7 on the APC membrane and/or because of some
participation of other costimulatory molecules, the blocking peptide
did not completely abrogate the costimulatory effect of BALMs.
Similarly, in the original studies describing the activity of this B7
homolog, the costimulatory signals were also not completely inhibited
(24, 25). Other costimulatory molecules such as ICAM 1
(CD54)-LFA 1 (CD11b) and LFA3-CD2, which would not be affected by that
peptide, may also contribute.
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Having shown that BALMs from
CFTR-/- and IL-10-/-
mice had similarly increased B7 expression and T cell costimulatory
activity, we sought to further strengthen the link between these two
deficiencies by determining whether administration of exogenous IL-10
to the CFTR-/- mice would make their BALMs more
like those from their +/+ counterparts. We treated eight
CFTR-/- mice with rmIL-10 and control groups of
CFTR-/- and CFTR+/+ mice
with buffer alone (eight mice in every group). All mice were sacrificed
on day 14, and two pools of BALMs from four mice each were made for
each treatment category, so all subsequent determinations could be done
on replicates. The IL-10 treatment did not appear to have any adverse
effect, and CFTR-/- mice at this age do not
have any visible pathologic changes in their lungs or any evident
inflammatory bowel disease. As shown in Fig. 6A and Table II, BALMs from control
(CFTR+/+) mice had minimal B7 expression. In
contrast, cells from CFTR-/- mice treated with
buffer alone were >86% positive for B7-1, with mean fluorescence
intensity of 66.0 (Fig. 6B). When
CFTR-/- mice were treated with 1 µg of
rmIL-10 each day for 2 wk, their BALMs had much lower B7-1 expression
(Fig. 6C; 19.9% positive, mean fluorescence intensity of
13.5, p < 0.05) than CFTR-/-
mice treated with vehicle alone. The B7-2 was also decreased by the
IL-10 treatment, but there were no significant changes in MHC-II
expression (Table II). The overall B7 expression on BALMs from the
CFTR-/- mice treated with vehicle only was
somewhat higher than on the cells shown in Fig. 3
and Table I. We think
that daily handling and exposure to BSA in the treatment vehicle may be
responsible for the increased B7 on BALM expression in this set of
experiments.
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and
show that the rmIL-10 treatment (right pair of bars)
decreased the costimulatory activity of the BALMs from the
vehicle-treated CFTR-/- animals (center
pair of bars), just as it had decreased the B7 expression. We
speculate that the higher activity seen with cells from
CFTR+/+ and CFTR-/- mice
treated with vehicle alone in these experiments compared with those
shown in Fig. 5, A and B, may be due to the daily
exposure to BSA in the control vehicle and then culture in the presence
of FCS.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In our studies of the functional significance of the IL-10 deficiency in the BAL fluid of CF patients, we observed that B7 expression on airway macrophages from the patients was increased. Because the lungs of most CF patients are chronically infected, we could not determine whether the increased B7 expression was directly related to the CFTR defect or the chronic infection. Therefore, we used uninfected CFTR-/- and IL-10-/- mice as models. Whereas CFTR+/+ mice had detectable IL-10 in their concentrated BAL fluid, CFTR-/- mice were deficient in IL-10 production. Because none of these experimental animals had evidence of infection, we suggest that CFTR malfunction may be directly associated with inhibition of IL-10 production, though the exact mechanism is unknown. Thus, we suggest that the defective CFTR function in the lungs (28) causes differentiation of the BALMs in an environment deficient in IL-10 and that this in turn increases expression and function of T cell costimulatory molecules, which may serve as a biologic marker of the lack of the IL-10.
The lack of IL-10 in the BAL fluid from untreated CFTR-deficient mice might also explain the observation that CFTR-/- mice given experimental endobronchial P. aeruginosa infection have increased levels of proinflammatory cytokines, increased neutrophil influx into the lung, increased weight loss, and increased mortality compared with their wild-type littermates (29). Comparable changes were seen in IL-10-/- vs wild-type littermates with the same or similar model infection (13, 30, 31). Thus, if IL-10 is absent or decreased, as in CF, besides an increased inflammatory response to infectious stimuli, the B7 expression may be constitutively increased. Other investigators have shown that BALMs isolated from patients with sarcoidosis and other chronic inflammatory lung diseases also have increased expression of the costimulatory molecule B7 on their surfaces (32, 33, 34). Those data demonstrate that BALMs from patients with a variety of diseases possess the capability to act as competent accessory cells and that this accessory function is at least in part mediated by the expression of CD80. Although it is not known whether there is a lack of endogenous IL-10 in those conditions, it has been shown that the increased accessory function or severity of inflammatory response can be reversed by IL-10 (35, 36).
As a model of IL-10 deficiency in CF lung, we examined IL-10-/- mice and compared them with CFTR-/- mice. Clearly, both IL-10-/- and CFTR-/- mice have increased expression and function of B7-1 and B7-2 compared with their +/+ counterparts. The striking similarity of the results for the IL-10-/- and CFTR-/- support the hypothesis that defects in CFTR lead to deficiency of IL-10 in the lung and that the increased B7 expression and function result from this IL-10 deficiency. To further test our hypotheses on the effects of IL-10 deficiency in the CF lung, we administered exogenous IL-10 to the CFTR-/- mice to determine whether this could modify the expression of costimulatory molecules and their increased function observed with their BALMs. The results clearly show that the B7 expression was reduced but not totally normalized by this treatment and that the costimulatory activity of lung macrophages was also reduced when the B7 expression was decreased by this treatment, adding further support to our hypothesis that the IL-10 deficiency in the CF bears responsibility for these differences from the normal lung.
Our results, which show only an incomplete effect of 14 d of treatment with IL-10, may be consistent with those of Chelen et al. (14), who reported that addition of anti-IL-10 mAb to in vitro cultures of alveolar macrophages for less than 24 h did not enhance their expression of B7-1 costimulatory molecules and suggest that prolonged exposure to the tissue microenvironment determines the macrophage’s phenotype (37, 38). We speculate that the increased B7 expression and function on BALMs in the CF lung, which can clearly augment the Ag-presenting capabilities of the macrophages, may contribute to the excessive Ab responses against P. aeruginosa and the hyperglobulinemia seen in most CF patients(7, 39, 40).
The use of the synthetic peptide that blocks the binding site on CD28 for B7 clearly shows that the costimulatory activity of the BALMs from the IL-10-/- and CFTR-/- mice is highly dependent on their B7 expression. Splenic APCs from CFTR-/- and CFTR+/+ did not differ significantly in function, and splenic APCs from all of these types of mice appeared less dependent than BALMs on B7 for their costimulatory activity, perhaps because they can use other molecules to serve this function (41, 42). Although we did not measure tissue-specific levels of IL-10 in the spleen, the costimulatory activity of splenic APCs in the CFTR-/- mice was less affected by the IL-10 treatment than were the lung macrophages. This is consistent with the idea that airway epithelial cells, which are the likely source of the IL-10 in the normal lung (1, 2), may be more dependent on CFTR for their normal function, including IL-10 production, than many other types of cells. Lymphocytic cell lines and lymphoblasts have been shown to express CFTR, and T cell lines from CF patients have been shown to produce less IL-10 than comparable cells from healthy individuals (43, 44). However, those differences are not as great as the differences in IL-10 content of BAL documented in CF vs normal patients and CFTR-/- vs CFTR+/+ mice. This also supports the conclusion that IL-10 production in the airways is more affected by the defects in CFTR because the airway epithelial cells are a major source of its production.
Our data support the hypothesis that the bronchial epithelium plays a significant role in defining the local immune response and may illustrate an important mechanism by which a local tissue controls its immunologic milieu. We propose that IL-10 is a an essential mediator of this regulation and that local IL-10 deficiency may contribute, not only to increased costimulatory activity for Ags in the airways, but also to other pathogenic and experimental features of CF lung disease. Furthermore, the data show that B7 expression on bronchoalveolar macrophages may serve as an important biological marker demonstrating the functional significance of the constitutive IL-10 secretion in the normal lung in vivo and, conversely, its absence in the CF lung. The increased expression of B7 on CF lung macrophages may have serious consequences for gene therapy because increased Ag-presenting activity may lead to enhanced immune responses against the various vectors or transfected cells. Reversal of immunoregulatory abnormalities caused by insufficient IL-10 (45) might be considered for future therapeutic development.
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Acknowledgments |
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Footnotes |
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2 Current address: Department of Pulmonary and Critical Care Medicine, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195.
3 Address correspondence and reprint requests to Dr. Melvin Berger, Department of Pediatrics, Case Western Reserve University, 2101 Adelbert Road, Cleveland, OH 44106. E-mail address: mxb12{at}po.cwru.edu
4 Abbreviations used in this paper: BAL, bronchoalveolar lavage; CF, cystic fibrosis; BALM, bronchoalveolar lavage macrophage; MHC-II, MHC class II; CFTR, cystic fibrosis transmembrane conductance regulator; rm, recombinant mouse.
Received for publication June 18, 2001. Accepted for publication November 30, 2001.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, IL-8, soluble ICAM-1, and neutrophils in sputum of cystic fibrosis patients. Pediatr. Pulmonol. 21:11.[Medline]
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 O. Tabary, C. Muselet, S. Escotte, F. Antonicelli, D. Hubert, D. Dusser, and J. Jacquot Interleukin-10 Inhibits Elevated Chemokine Interleukin-8 and Regulated on Activation Normal T Cell Expressed and Secreted Production in Cystic Fibrosis Bronchial Epithelial Cells by Targeting the IkB Kinase {alpha}/{beta} Complex Am. J. Pathol., January 1, 2003; 162(1): 293 - 302. [Abstract] [Full Text] [PDF]
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