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Allergy and Immunology Division, University of Pennsylvania School of Medicine, Philadelphia, PA 19104
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Therefore, eosinophils may play a key role in the determination of the
unique cytokine microenvironment within the granuloma milieu. These
studies investigated the potential role of eosinophils in the
regulation of granuloma immunopathology. We have characterized spleen-
and granuloma-derived eosinophils based on cellular activation and
cytokine production during the development of murine schistosomiasis.
Based on the criteria of hypodensity and CD69 expression, granuloma
eosinophils were highly activated and very homogeneous at 7 and 11 wk
postinfection. Splenic eosinophils were also activated at 7 wk
postinfection, but were much more heterogeneous than their granuloma
counterparts. By 11 wk postinfection, few hypodense splenic eosinophils
were observed. Eosinophils represented the majority of
cytokine-producing cells in the granuloma and were a dominant source of
IL-4. Eosinophils also produced IL-2, IL-5, and IFN-, using the
criteria of mRNA in situ hybridization and intracellular cytokine
staining by FACS. Granuloma eosinophil activation and cytokine
production were greatest at the time of maximum granuloma formation,
i.e., 10–12 wk after initial cercarial exposure. Therefore, locally
activated eosinophils, not Th2 lymphocytes, produce the majority of Th2
cytokines in the granuloma milieu and may be important determinators of
immunopathology in schistosomiasis. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The mechanisms controlling granuloma formation and modulation have been the focus of intense research over the past 30 yr. Granulomas are generated and modulated by T cells and alterations in the cytokine milieu. Granuloma formation is known to be a CD4+-dependent, cell-mediated process, as evidenced by the suppression of hepatic granuloma formation in anti-CD4-treated mice (6, 7) and the formation of smaller granulomas in athymic mice (8). In addition, granuloma formation can be adoptively transferred by T cells (3).
The importance of Th1 and Th2 cytokines produced during granuloma formation is complex and controversial (9, 10, 11). The dominance of IL-4 and IL-5 as the disease progresses has largely been attributed to the effects of soluble egg Ag, which is produced with the onset of egg deposition. The majority of studies that have analyzed cytokine production have ascribed the cytokine patterns to the activities of T cell subpopulations and monocytes. However, these studies used in vitro activated cell cultures or tissue extraction techniques that could not identify with certainty the exact nature of the cells that actually produced the cytokines or quantitate the production of these cytokines on a per cell basis.
Eosinophils are also a prominent cell within schistosome granulomas and are known to produce a variety of cytokines (12). Therefore, in the present study we have examined the activation of eosinophils and the production of specific cytokines and have combined this information with morphological characterization to precisely define the cellular source(s) of cytokine in the spleen as well as in hepatic granulomas. Our data show conclusively that eosinophils are the dominant source of IL-4 during Schistosoma mansoni infection and that eosinophils are the cell population most responsible for the cytokine profiles that had been previously attributed to a Th1 to Th2 shift.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Female C57BL/6 (Harlan Sprague-Dawley, Indianapolis, IN) were used in all experiments. Mice (5–6 wk of age) were infected with 30 cercariae percutaneously by immersion of the tail for 1 h in water containing S. mansoni cercariae according to the method of Bruce and Radke (13). Infected snails were obtained from Fred Lewis at the Biomedical Research Institute (Rockville, MD).
Cell preparation
Spleens and livers were aseptically removed from infected animals, and cell suspensions were prepared as previously described (14).
Cytokine ELISPOT assay
ELISPOT4 assays were
performed based on the method described by Klinman and Nutman (15).
Immulon II plates (Dynatech, Chantilly, VA) were coated overnight at
4°C with 1 µg of capture Ab (anti-IFN- clone HB170 and
anti-IL-4 clone 11B11, PharMingen, San Diego, CA). Plates were
washed three times with PBS/Tween-20 (0.5%) and blocked with 200
µl/well of RPMI and 5% FCS for 2 h at 37°C. Single cell
suspensions were prepared from spleen and granulomas as described
above, except for the omission of RBC lysis. Cells were suspended in
complete medium at a density of 1 x 106
leukocytes/ml. One hundred microliters of fivefold serial dilutions of
cells were added to previously aspirated triplicate wells. Plates were
incubated for 3 h at 37°C under 5% CO2. Following
incubation, the plates were washed three times with PBS followed by
three additional washes with PBS/Tween. Biotinylated
anti-cytokine-detecting Abs (anti-IFN- clone XMG-6 and
anti-IL-4 clone BVD6, PharMingen) were diluted to 4 µg/ml in
PBS/Tween, and 100 µl was added per well. Plates were incubated for
1 h at 37°C under 5% CO2 and then washed three
times with PBS and three times with PBS/Tween. Streptavidin-alkaline
phosphatase (Jackson ImmunoResearch Laboratories, West Grove, PA),
diluted 1/2000 in PBS/Tween-20 and 5% FCS (100 µl/well), was added,
and the plates were incubated 30 min at 37°C in 5% CO2.
Plates were washed five times with PBS/Tween, and 100 µl of BCIP
(Sigma, St. Louis, MO) at 1 mg/ml in AMP buffer (0.15 M
2-amino-2-methyl-1-propanol) containing 0.6% agarose was added per
well. The plates were allowed to stand until the agarose had set, were
stored at room temperature overnight, and were scored the next day. All
scoring was performed by a single investigator in a coded manner.
Flow cytometry
Surface and intracellular cytokine staining.
Freshly prepared cell suspensions (1 x 107/ml)
were incubated in RPMI 1640 and 10% FCS with 10 µg/ml brefeldin A
(Sigma) for 3 h at 37°C in 5% CO2. Following
incubation, the cells were harvested, surface stained (PharMingen 
CD69 clone H1.2F3), and fixed overnight in 2.5% paraformaldehyde. The
following day, cells were stained for intracellular cytokines as
previously described (14) using FITC-conjugated anti-cytokine mAbs
(PharMingen; IL-2 clone 54B6, IL-4 clone 11B11, and IFN-
clone XMG1.2). After staining, the samples were immediately analyzed by
FACS. All data were acquired and analyzed using a FACScan or
FACScalibur instrument equipped with CellQuest software (version 3.0.1)
or FACStar Plus equipped with LYSIS II software (Becton Dickinson,
Mountain View, CA).
Cell sorting. Unlabeled cell suspensions prepared from spleen or granulomas were sorted on a FACStar Plus instrument. Lymphocyte and eosinophil gates were defined by forward and side light scatter parameters, and sorted cells were stained with Biebrich Scarlet and hematoxylin to assess cell phenotype and purity in each gate. Sorted cells were also used in mRNA in situ hybridization experiments.
In situ mRNA hybridization
Oligonucleotide probes were labeled with digoxigenin
(DIG)-11–2',3'-dideoxy-UTP using a Genius 5 kit (Boehringer Mannheim,
Indianapolis, IN). The sequences of the antisense probes used were as
follows: IL-2, 5'-GAGTCAAATCCAGAACATGC-3'; IL-4, 5'-
AACGCTACACACTGCATCTT-3'; IL-5, 5'-GAACTCTTGCAGGTA ATCCA-3'; and
IFN-, 5'-GACTTCAAAGAGTCTGAGGT-3'. Sorted cells (0.1 x
106) were cytospun and mounted on SuperFrost Plus-precoated
slides (Fisher, Pittsburgh, PA). The slides were fixed in 4%
paraformaldehyde (pH 7.4) for 20 min, washed once with
diethylpyrocarbonate/PBS, and incubated with acetylation solution (0.1
M triethanolamine and 0.25% acetic anhydride) for 20 min at room
temperature in a humid chamber. The slides were then washed and
incubated in 70% ethanol for 1 h at 4°C before two washes with
2x SSC. Prehybridization solution was added (per 1 l:50 mg of
dextran sulfate, 100 ml of 20x SSC, 50 ml of deionized formamide, 2 ml
of 0.5 M EDTA (pH 8.0), 1 mM Tris (pH 8.0), 100 mg of denatured sheared
salmon sperm DNA, and 20 ml of 50x Denhardt’s solution), and the
slides were incubated for 1 h at 37°C in a humidified chamber.
Hybridization solution (prehybridization solution plus DIG-labeled
probe) was then added, and the samples were incubated overnight at
45°C in a humid chamber. Slides were washed three times with 2x SSC
and then washed four times with 0.1x SSC, incubating each wash for 15
min at 37°C in a humid chamber, followed by one additional 15-min
wash in double distilled-H2O. Blocking buffer (1%
blocking reagent, 0.1 M malic acid, and 0.15 M NaCl, pH 7.5) was added
to each slide, and the slides were incubated for 1 h at room
temperature in a humid chamber followed by addition of alkaline
phosphatase-conjugated anti-DIG (1/1000; Genius 3 Kit,
Boehringer Mannheim) and incubation overnight at 4°C in a humid
chamber. The slides were washed three times for 10 min each time at
room temperature in buffer 3 (100 mM Tris-HCl, 100 mM NaCl, and 50 mM
MgCl2, pH 9.5) and dried. Slides were covered with 4-nitro
blue tetrazolium chloride/BCIP substrate (3.5 µl BCIP, 4.5 µl
4-nitro blue tetrazolium chloride, and 1 ml buffer 3), incubated in the
dark for 2–4 h at room temperature until color developed, and washed
with H2O. Nuclei were counterstained with methyl green, and
the slides were mounted with Permount (Fisher). Negative controls
included cytospun samples incubated with either no probe or irrelevant
antisense probes.
Determination of eosinophil density
Eosinophil density was determined using the method of Hua et al. (16). Cells were isolated from the spleens and granulomas of 7- or 11-wk infected animals as described above for spleen cell preparation without collagenase treatment. Discontinuous Percoll (Sigma) density gradients were prepared by mixing heavy and light solutions (pH 7.4) and weighing 2 ml of individual solutions as described by Gartner (17) to determine the actual density of each solution. Gradients were composed of 1.5 ml of each Percoll solution, layered sequentially in a 15-ml polypropylene tube at densities of 1.09, 1.085, 1.08, 1.075, 1.07, 1.065, and 1.06 g/ml. Cells (0.5–5 x 107) were suspended in 1 ml of Percoll (1.055 g/ml) and layered on top of each gradient. After centrifugation in a fixed angle rotor at 2500 x g for 30 min at room temperature, cells were removed from each interface of the gradient and washed twice. Eosinophils were stained with Hinkelman’s solution (0.5% eosin Y, 0.5% formalin, and 95% phenol) and enumerated in a hemacytometer.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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We initially wished to determine the overall profile of total
cytokine-producing cells in the spleens and granulomas of S.
mansoni-infected mice and to compare that profile to the actual
kinetics of granuloma formation. The numbers of ELISPOTs per 100,000
spleen- or granuloma-derived cells harvested from animals 6–17 wk
postinfection are shown in Figs. 1
and
2, respectively. The numbers of
IFN--producing ELISPOTs counted per 100,000 cells were similar for
spleen- and granuloma-derived cells obtained from acutely infected
animals (
20/105). A slight decline in the number of
IFN--producing cells and a slight increase in the number of
IL-4-producing cells were noted in splenic populations as the disease
progressed to 15–17 wk postinfection. Conversely, dramatic differences
were observed in the numbers of IL-4-producing ELISPOTs per 100,00
cells when spleen and granuloma cells were compared. Very few IL-4
ELISPOTs were observed in the spleen (<20/100,000 cells) at all times
of infection examined. In contrast, very large numbers of
IL-4-producing ELISPOTs were obtained with granuloma cells at all
periods of infection (note the change in scale). In addition, the
number of cytokine-producing cells, as a function of weeks
postinfection, correlated well with our previously published kinetics
of granuloma formation (18). The number of IL-4-producing ELISPOTs
peaked around 6,500/100,000 at 13 wk postinfection. This frequency per
se exceeded the total frequency of all lymphocytes in the granulomas
(4.03 ± 3.08%). Additional studies had previously shown that
only a minority of lymphocytes in the granuloma produced IL-4 (14).
These findings suggested that Th2 cytokines were being produced by
large numbers of granuloma cells that were not lymphocytes.
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We next wished to determine the exact nature of the cells that
were responsible for the production of cytokines. Spleen and granuloma
cells from acutely infected animals were sorted by forward and side
light scatter characteristics (Fig. 3).
The cellular gates defined by forward and side light scatter
characteristics were the same as those used for cell sorting and
intracellular cytokine staining experiments. Sorted cells were stained
with hematoxylin and Biebrich Scarlet, which specifically binds the
arginine-rich proteins of eosinophil granules, but not neutrophil
granules (19, 20), to assess purity and morphology. The results are
shown in Table I. Cells recovered from
the lymphocyte gates in the spleen and granuloma were 98 and 89% pure
small mononuclear lymphocytes, respectively. Eighty-five percent of the
cells within the granuloma eosinophil gate were eosinophils as
evidenced by Biebrich Scarlet staining and the presence of a
donut-shaped nucleus, which is highly characteristic of murine
eosinophils. Seventy-six percent of the splenic eosinophil-gated cells
were eosinophils by morphological and staining criteria. The lower
percent purity observed in the splenic eosinophil gate was due to the
presence of polymorphonuclear cells and large mononuclear cells with
extensive cytoplasm, most likely monocytes or macrophages. The number
of contaminating lymphocytes in the eosinophil gate was <5%.
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. At 8–12 wk postinfection, the spleen
cell preparation was rich in lymphocytes, and eosinophils constituted
less than 20% of the total cells. In contrast, the granuloma cell
preparations from these acutely infected animals were composed of
approximately 90% eosinophils. The striking difference in the cellular
compositions of the spleen and granulomas in S.
mansoni-infected mice along with ELISPOT data showing high
numbers of IL-4-producing ELISPOTs in the granuloma suggested that
eosinophils might be the dominant numerical source of IL-4-producing
cells. Therefore, we studied the characteristics of these eosinophils
in more detail.
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Eosinophil activation.
First, we examined the relative state of activation of splenic and
granuloma eosinophils. To measure one criteria of physiological
activation, we determined the cellular densities of splenic and
granuloma eosinophils. One characteristic of activated eosinophils is a
decrease in relative density due to a combination of cellular
enlargement and loss of intracellular granules secondary to
degranulation (16, 21). The density of blood eosinophils isolated from
normal, noninfected mice is between 1.070–1.080 g/ml (16). Fig. 4A displays the density
profile of splenic and granuloma eosinophils isolated 7 wk
postinfection during the earliest phase of granuloma formation. The
broader, higher density curve observed for splenic eosinophils
indicated that these cells were very heterogeneous with regard to
cellular density and were nonactivated. Approximately 80% of the
splenic eosinophils were recovered from the density interfaces between
1.065, 1.070, and 1.075 g/ml. The granuloma eosinophils were hypodense
and were more homogeneous, with 70% of the eosinophils
accumulating at the 1.065 g/ml interface. The density determinations
were repeated at 11 wk postinfection, during the time of peak granuloma
formation. As shown in Fig. 4B, the splenic eosinophils were
of normal density and were much more homogeneous at 11 wk postinfection
than at 7 wk postinfection. Over 50% of the total eosinophils
accumulated at the 1.075 g/ml interface. Similar to the 7 wk point, the
granuloma-derived eosinophils were highly activated, i.e., homogeneous
and of lower density than splenic eosinophils.
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Eosinophil cytokine production. Previous studies and our initial observation had indicated that high levels of cytokines are produced in the granulomas. We wished to assess the exact source of these cytokines with specific reference to eosinophils.
Detection of eosinophil cytokine mRNA by in situ hybridization
We first directly demonstrated that eosinophils potentially could
produce cytokines by measuring cytokine mRNA in these cells. Sorted
spleen and granuloma eosinophils from 7-, 9-, 11-, and 17-wk-infected
animals were cytospun, and IL-2, IL-4, IL-5, and IFN- mRNA were
detected by in situ hybridization. Negative controls shown in Fig. 5 were incubated with no probe.
Additional samples were incubated with irrelevant antisense oligo
probes and were also negative for cytokine mRNA (data not shown).
Splenic and granuloma eosinophils obtained from mice 7 wk postinfection
stained brightly for mRNA encoding IL-2, IL-5, and IL-4 (Fig. 5, A and B). IFN- mRNA staining was less
intense at 7 wk postinfection, particularly in granuloma eosinophils.
At 11 wk postinfection, the intensity of IL-2 and IFN- mRNA staining
in splenic eosinophils increased, and staining of IL-4 and IL-5 mRNA
remained strong (Fig. 5C). mRNA staining for IL-2, IL-4, and
IL-5 was slightly less intense in granuloma eosinophils at 11 wk
postinfection (Fig. 5D) than at 7 wk, while IFN- mRNA
staining increased.
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mRNA staining demonstrated unequivocally that eosinophils had the
potential to produce cytokines. However, these initial in situ studies
were not quantitative and did not prove that the eosinophils actually
spontaneously produced cytokine proteins in vivo. Therefore, we
measured intracellular cytokine protein production by freshly isolated
cells using flow cytometry. Fig. 6 shows
typical histogram profiles for eosinophils isolated from spleens and
granulomas of S. mansoni-infected mice, 9 wk after cercarial
exposure. The cells were cultured for 3 h without the addition of
any in vitro activation agents but in the presence of brefeldin A.
Brefeldin A interferes with protein secretion; therefore, any cytokine
protein that was being produced in vivo, just before isolation of the
cells, accumulates within the cell and can be detected by flow
cytometric analysis.
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, A and
B). Low levels of IFN- protein were detected in both
splenic and granuloma eosinophils in this experiment (Fig. 6, C and D). In replicate experiments, IFN-
staining was slightly more intense. However, IFN- staining was
consistently less than IL-2 staining in every experiment. This low
level of IFN- expression was consistent with the numbers of
IFN-+ ELISPOTs detected (Fig. 1). Splenic eosinophils
were only slightly positive for IL-4. Granuloma eosinophils, on the
other hand, were highly positive for IL-4 protein as indicated by the
shift of nearly an entire log unit in the mean channel fluorescence
intensity of the entire eosinophil population in the IL-4-stained
sample compared with the isotype control (Fig. 6, E and
F).
Splenic and granuloma lymphocyte cytokine production were compared.
IL-4 data are presented in Fig. 6. Splenic lymphocytes produced
virtually no IL-4 protein based on similar isotype and IL-4 cytokine
fluorescence histograms (Fig. 6G). A small percentage of
granuloma lymphocytes (5%) produced modest amounts of IL-4 (Fig. 6H). Since the same concentration of fluorochrome-labeled Ab
was added to each sample, and all other conditions were held constant,
the relative change in fluorescence intensity of cytokine-stained
samples compared with isotype controls is a semiquantitative, but
proportional, measurement of intracellular cytokine concentration.
Based on the high percentages of eosinophils producing IL-4 and their
numerical dominance in the granuloma (90% of the granuloma cell
preparation), these data demonstrated that granuloma lymphocytes were
producing a small amount of IL-4 and that the eosinophils were the
dominant quantitative source of IL-4 in the granuloma.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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, while IL-4 and IL-5 are the dominant
cytokines in the largest granulomas (25, 26, 27). The majority of studies
to date have analyzed cytokine production by ELISA or
immunocytochemical techniques after in vitro stimulation. Although
these studies were useful in characterizing the general cytokine
responses elicited as a result of S. mansoni infection, they
provided limited information about the nature of the cell population(s)
that was specifically producing cytokines in vivo. In addition, the
studies introduced a potential confounding variable, i.e., the effects
of in vitro incubation with lectins or Ag. We chose to examine in
greater detail the cell types responsible for the observed Th2 cytokine
response and in particular to focus on eosinophils immediately after
their isolation from infected mice. Our initial ELISPOT studies with spleen and granuloma cells showed large numbers of IL-4-producing cells in the granuloma compared with those in the spleen. Several types of cells could contribute to cytokine production. A recent study suggested that non-CD4+ peritoneal exudate cells produced IL-4 following i.p. injection of S. mansoni eggs. Immunocytochemical analyses of these cells suggested that they were eosinophils (28). In the present study we assessed the nature of the cells responsible for IL-4 production within hepatic granulomas.
CD4+ T cells are required for granuloma formation and have
been shown to produce cytokines within granulomas (14). For example,
both spleen and granuloma lymphocytes produce IL-4 (Fig. 6, G and H). However, in this study we show that
eosinophils constitute approximately 90% of the cells in acute
granulomas and are highly positive for IL-4. Therefore, eosinophils
represent the dominant source of IL-4 in granulomas, and eosinophils
may be the actual source of the observed Th1 to Th2 cytokine shift seen
in schistosomiasis.
Eosinophils are thought to play a major role in a variety of human diseases, including allergic inflammation, malignancy, and host defense against helminth infections (29). However, the exact role(s) of eosinophils in schistosomiasis relative to immunopathology remains unclear.
Eosinophils may participate in Ab-dependent protective immune responses (30, 31). Human eosinophils express IgE receptors that participate in an IL-5-dependent Ab-dependent cell-mediated cytotoxicity reaction against schistosomula in vitro (32, 33, 34, 35). Eosinophils also mediate the destruction of miracidia and schistosome eggs (30). Murine eosinophils do not express surface IgE receptors, but do express IgG receptors. An oxidative burst can be triggered through these IgG receptors (30), suggesting that murine eosinophils may use an alternate mechanism to eliminate schistosomula.
To further define the role of murine eosinophils in the pathology of S. mansoni infection, we examined several parameters, including activation and cytokine production. During the acute phase of infection, we showed selective degranulation of granuloma eosinophils and very heterogeneous splenic eosinophils. As disease progressed, granuloma eosinophils remained hypodense, and splenic eosinophils became much more homogeneous and normal. In additional experiments, we examined expression of the early activation marker CD69 on splenic and granuloma eosinophils. Based on greater mean channel fluorescence intensities, we detected enhanced levels of CD69 expression on granuloma eosinophils compared with splenic eosinophils (data not shown). These findings suggested partial activation of eosinophils in the spleen and augmented eosinophil activation in the granuloma, perhaps due to greater Ag interactions in the granuloma.
Granuloma eosinophils produce a variety of cytokines. Of special interest is the production of the Th2-type cytokines IL-4 and IL-5 by granuloma eosinophils. Several investigators have shown that a Th1 to Th2 shift has profound effects on the ability of the host to respond to other infectious diseases, specific Ags, and allergens (36, 37, 38). Flow cytometry data for intracellular IL-5 production, a more stringent marker of the Th2 response, were not presented in this report because we had technical difficulty detecting intracellular IL-5 by flow cytometry. This was most likely due to the fact that we did not stimulate freshly isolated splenic and granuloma cell suspensions in vitro. However, we did demonstrate by in situ hybridization that sorted splenic and granuloma eosinophils had high levels of IL-5 mRNA expression. Anti-IL-5 treatment has been shown to specifically deplete eosinophil accumulation in granulomas (39), further supporting the importance of IL-5 in eosinophil recruitment. Ongoing RNase protection studies using sorted splenic and granuloma lymphocytes and eosinophils will further define which cells are producing specific cytokine and chemokine RNAs. The kinetics of cytokine and chemokine production will help determine whether expression is being regulated in an autocrine, paracrine, or juxtacrine manner. The identification of eosinophils as the dominant contributors to the Th2 response in S. mansoni infection suggests that other studies addressing Th1 and Th2 responses may need to be reinterpreted to determine the cellular source(s) of cytokines and the basis of systemic perturbations of immunity.
Of particular interest is why eosinophils achieve such numerical dominance in granulomas and what is the implication of that dominance. Eosinophils may only reach the granuloma as a result of their response to factors produced by Ag-specific T cells. Indeed, anti-IL-5 treatment will prevent the accumulation of eosinophils in granulomas but does not grossly affect granuloma formation per se (39). Conversely, once the eosinophils have reached the granulomas, they may produce cytokines that adversely affect T cells, especially pathogenic Th1 cells, but that augment their own survival. Our preliminary data suggest that, unlike lymphocytes, eosinophils do not undergo apoptosis in the granulomas. Perhaps this survival is augmented by the presence of large amounts of IL-4 and IL-5, which have been shown to promote eosinophil survival in vitro. Finally, eosinophils have been shown to produce a number of proteases, peroxidases, histamines, etc., which may directly affect local inflammation and thereby effect pathology. The mechanisms of cytokine-regulated survival and apoptosis by granuloma subpopulations has profound implications vis-à-vis pathology and are the subject of current study in our laboratory.
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Footnotes |
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2 Current address: Department of Parasitology, Akita University School of Medicine, Akita, Japan.
3 Address correspondence and reprint requests to Dr. S. Michael Phillips, Allergy and Immunology Division, University of Pennsylvania School of Medicine, 518 Johnson Pavilion, Philadelphia, PA 19104-6057. E-mail address:
4 Abbreviations used in this paper: ELISPOT, enzyme-linked immunospot; BCIP, 5-bromo-4-chloro-3-indolyl-phosphate, toluidine salt; DIG, digoxigenin.
Received for publication July 10, 1998. Accepted for publication September 29, 1998.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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mRNA, and IL-2 mRNA levels in the granulomatous livers, mesenteric lymph nodes, and spleens during the course of modulation. J. Immunol. 148:2261.[Abstract]
RII (CD32) is linked to apoptotic pathways in murine granulocyte precursors and mature eosinophils. Blood 90:1267.This article has been cited by other articles:
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  S. I. Ochkur, E. A. Jacobsen, C. A. Protheroe, T. L. Biechele, R. S. Pero, M. P. McGarry, H. Wang, K. R. O'Neill, D. C. Colbert, T. V. Colby, et al. Coexpression of IL-5 and Eotaxin-2 in Mice Creates an Eosinophil-Dependent Model of Respiratory Inflammation with Characteristics of Severe Asthma J. Immunol., June 15, 2007; 178(12): 7879 - 7889. [Abstract] [Full Text] [PDF]
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 R. M. Reiman, R. W. Thompson, C. G. Feng, D. Hari, R. Knight, A. W. Cheever, H. F. Rosenberg, and T. A. Wynn Interleukin-5 (IL-5) Augments the Progression of Liver Fibrosis by Regulating IL-13 Activity Infect. Immun., March 1, 2006; 74(3): 1471 - 1479. [Abstract] [Full Text] [PDF]
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A. Gessner, K. Mohrs, and M. Mohrs Mast Cells, Basophils, and Eosinophils Acquire Constitutive IL-4 and IL-13 Transcripts during Lineage Differentiation That Are Sufficient for Rapid Cytokine Production J. Immunol., January 15, 2005; 174(2): 1063 - 1072. [Abstract] [Full Text] [PDF]
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A. B. Stavitsky Regulation of Granulomatous Inflammation in Experimental Models of Schistosomiasis Infect. Immun., January 1, 2004; 72(1): 1 - 12. [Full Text] [PDF]
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L. H. Hogan, M. Wang, M. Suresh, D. O. Co, J. V. Weinstock, and M. Sandor CD4+ TCR Repertoire Heterogeneity in Schistosoma mansoni-Induced Granulomas J. Immunol., December 1, 2002; 169(11): 6386 - 6393. [Abstract] [Full Text] [PDF]
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C. Jakubzick, S. L. Kunkel, B. H. Joshi, R. K. Puri, and C. M. Hogaboam Interleukin-13 Fusion Cytotoxin Arrests Schistosoma mansoni Egg-Induced Pulmonary Granuloma Formation in Mice Am. J. Pathol., October 1, 2002; 161(4): 1283 - 1297. [Abstract] [Full Text] [PDF]
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C. Faveeuw, V. Angeli, J. Fontaine, C. Maliszewski, A. Capron, L. Van Kaer, M. Moser, M. Capron, and F. Trottein Antigen Presentation by CD1d Contributes to the Amplification of Th2 Responses to Schistosoma mansoni Glycoconjugates in Mice J. Immunol., July 15, 2002; 169(2): 906 - 912. [Abstract] [Full Text] [PDF]
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 C. F. Lucchinetti, R. N. Mandler, D. McGavern, W. Bruck, G. Gleich, R. M. Ransohoff, C. Trebst, B. Weinshenker, D. Wingerchuk, J. E. Parisi, et al. A role for humoral mechanisms in the pathogenesis of Devic's neuromyelitis optica Brain, July 1, 2002; 125(7): 1450 - 1461. [Abstract] [Full Text] [PDF]
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 E L J van Rensen, R G Stirling, J Scheerens, K Staples, P J Sterk, P J Barnes, and K F Chung Evidence for systemic rather than pulmonary effects of interleukin-5 administration in asthma Thorax, December 1, 2001; 56(12): 935 - 940. [Abstract] [Full Text] [PDF]
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H. Kayaba, D. Dombrowicz, G. Woerly, J.-P. Papin, S. Loiseau, and M. Capron Human Eosinophils and Human High Affinity IgE Receptor Transgenic Mouse Eosinophils Express Low Levels of High Affinity IgE Receptor, but Release IL-10 upon Receptor Activation J. Immunol., July 15, 2001; 167(2): 995 - 1003. [Abstract] [Full Text] [PDF]
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 J. T. Kaifi, L. R. Hall, C. Diaz, J. Sypek, E. Diaconu, J. H. Lass, and E. Pearlman Impaired Eosinophil Recruitment to the Cornea in P-Selectin-Deficient Mice in Onchocerca volvulus Keratitis (River Blindness) Invest. Ophthalmol. Vis. Sci., November 1, 2000; 41(12): 3856 - 3861. [Abstract] [Full Text]
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D. C. Webb, A. N. J. McKenzie, A. M. L. Koskinen, M. Yang, J. Mattes, and P. S. Foster Integrated Signals Between IL-13, IL-4, and IL-5 Regulate Airways Hyperreactivity J. Immunol., July 1, 2000; 165(1): 108 - 113. [Abstract] [Full Text] [PDF]
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N. Hayashi, K. Matsui, H. Tsutsui, Y. Osada, R. T. Mohamed, H. Nakano, S.-i. Kashiwamura, Y. Hyodo, K. Takeda, S. Akira, et al. Kupffer Cells from Schistosoma mansoni-Infected Mice Participate in the Prompt Type 2 Differentiation of Hepatic T Cells in Response to Worm Antigens J. Immunol., December 15, 1999; 163(12): 6702 - 6711. [Abstract] [Full Text] [PDF]
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) or granulomas (
) of 7-wk
(A) or 11-wk (B) infected mice were
layered over a discontinuous Percoll gradient, and after
centrifugation, cells were recovered from each interface and counted as
described in Materials and Methods. Data are expressed
as the number of eosinophils recovered from each interface as a
percentage of the total number of eosinophils recovered from all
interfaces.
