Eotaxin/CCL11 is involved in acute, but not chronic, allergic airway responses to Aspergillus fumigatus

Jane M. Schuh, Kate Blease, Steven L. Kunkel, and Cory M. Hogaboam

Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan 48109


    ABSTRACT
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ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Eotaxin/CCL11 is a major chemoattractant for eosinophils and Th2 cells. As such, it represents an attractive target in the treatment of allergic disease. The present study addresses the role of eotaxin/CCL11 during acute and chronic allergic airway responses to the fungus Aspergillus fumigatus. Mice lacking the eotaxin gene (Eo-/-) and wild-type mice (Eo+/+) were sensitized to A. fumigatus and received either an intratracheal challenge with soluble A. fumigatus antigens (acute model) or an intratracheal challenge with live A. fumigatus spores or conidia (chronic model). Airway hyperresponsiveness and eosinophil, but not T cell, recruitment were significantly decreased at 24 h after the soluble allergen in A. fumigatus-sensitized Eo-/- mice compared with similarly sensitized Eo+/+ mice. In contrast, the development of chronic allergic airway disease due to A. fumigatus conidia was not altered by the lack of eotaxin. Together, these data suggest that eotaxin initiates allergic airway disease due to A. fumigatus, but this chemokine did not appear to contribute to the maintenance of A. fumigatus-induced allergic airway disease.

eosinophil; allergy; airway hyperreactivity


    INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

SINCE ITS DISCOVERY AND CLONING in a number of species (21, 36, 38), the CC chemokine eotaxin/CCL11 has received considerable research attention in the context of allergy and asthma (37). Although it is constitutively expressed in the lung, eotaxin/CCL11 levels are markedly increased in bronchoalveolar lavage (26, 29), sputum (46), and the airway wall (44, 48, 49) during asthmatic responses. Eotaxin/CCL11 appears to play a fundamental role in the development of allergic responses due, in large part, to its effects on the recruitment (10, 17, 39, 40) and activation of eosinophils (12, 35) and Th2 cells (30). Eotaxin/CCL11 appears to mediate this effect through the CC chemokine receptor-3 (11, 15), and a growing number of reports highlight the major therapeutic potential in targeting this chemokine receptor during the development of allergic airway inflammation (2).

Allergic responses to Aspergillus fumigatus are characterized by many features associated with asthma, including elevated IgE, enhanced Th2 cytokine levels, eosinophilia, bronchial hyperreactivity, and airway remodeling (22, 25). The eosinophil is a primary cellular culprit in the allergic pulmonary disease associated with this fungus (27), but few studies have addressed which factors control eosinophil recruitment into the airways during the course of this disease. The advent of murine models of A. fumigatus-induced pulmonary disease have facilitated study in this regard, and it has become apparent that chemokines are intricately involved in allergic airway disease due to A. fumigatus. For example, it has been shown that the introduction of soluble A. fumigatus antigens into the airways of mice previously sensitized to this fungus initiates an eosinophil-dominated peribronchial inflammation that persists for up to 48 h after the challenge and is dependent on the CC chemokine C10/CCL6 (20). More recently, we have begun to examine the impact of intratracheally introduced A. fumigatus spores (or conidia) on the persistence of allergic airway disease. It has been shown that CC chemokines that bind CCR1 (7) and CCR2 (6) have important roles in regulating the severity and features of this chronic fungal asthma model. Thus the purpose of the present study was to examine the role of eotaxin/CCL11 in the initiation and maintenance of A. fumigatus-induced asthmatic disease in mice.


    MATERIALS AND METHODS
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ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Murine models of acute and chronic fungal asthma. Specific pathogen-free (SPF) female eotaxin/CCL11 wild-type (Eo+/+) and eotaxin/CCL11 knockout (Eo-/-) mice were kindly provided by Dr. Rodrigo Bravo (Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ). The generation of Eo-/- mice was previously described in detail, and these mice do not exhibit phenotypic abnormalities (47). Both groups of mice were bred and maintained under SPF conditions in the University Laboratory Animal Medicine (ULAM) facility at the University of Michigan Medical School before and during experiments. Sensitization of mice to soluble A. fumigatus antigens was achieved using a previously described procedure (32, 33). Briefly, all mice received a total of 10 µg of A. fumigatus crude antigen (Greer Laboratories, Lenoir, NC) dissolved in 0.2 ml of incomplete Freund's adjuvant (Sigma Chemical, St. Louis, MO). Half of this preparation was then deposited in the peritoneal cavity, and the remainder was delivered subcutaneously. Two weeks later, mice received a total of 20 µg of A. fumigatus antigens dissolved in normal saline via the intranasal route. To initiate the acute fungal asthma model (20), mice received 20 µg of A. fumigatus antigen dissolved in normal saline via the intratracheal route 4 days after one intranasal challenge. To initiate the chronic fungal asthma model, A. fumigatus-sensitized Eo-/- and Eo+/+ mice received 5.0 × 106 A. fumigatus conidia suspended in 30 µl of 0.1% Tween 80 via the intratracheal route 7 days after the third intranasal challenge (19). Mouse lung responsiveness to intravenous methacholine administration and a number of additional parameters of allergic airway inflammation were examined at various times after the A. fumigatus intratracheal challenge in both models. Prior approval for mouse usage in these studies was obtained from the ULAM facility at the University of Michigan Medical School.

Measurement of bronchial hyperresponsiveness and lung harvest. Bronchial hyperresponsiveness in A. fumigatus-sensitized Eo+/+ and Eo-/- mice was measured in a Buxco plethysmograph (Buxco, Troy, NY) as previously described (19). Pentobarbital sodium (Butler, Columbus, OH; 0.04 mg/g of mouse body wt) was used to anesthetize each mouse before its intubation for ventilation with a Harvard pump ventilator (Harvard Apparatus, Reno, NV). After a baseline period in the Buxco mouse chamber, each mouse received 125 µg/kg of methacholine by tail vein injection, and airway responsiveness to this bronchoconstrictor was again calculated online. This dose of methacholine was selected to examine airway hyperresponsiveness in allergic mice because it failed to elicit a response in nonsensitized mice and consistently gave peak changes in airway resistance in A. fumigatus-sensitized mice. At the conclusion of the assessment of airway responsiveness, a bronchoalveolar lavage (BAL) was performed with 1 ml of normal saline. Approximately 500 µl of blood was then removed from each mouse and transferred to a microcentrifuge tube. Sera were obtained after the sample was centrifuged for 5 min. Whole lungs were finally dissected from each mouse and snap-frozen in liquid N2 or prepared for histological analysis.

Morphometric analysis of eosinophil and lymphocyte accumulation in BAL samples. Neutrophils, eosinophils, lymphocytes, and macrophages were enumerated in BAL samples cytospun (Shandon Scientific, Runcorn, UK) onto coded microscope slides. Each slide was stained with a Wright-Giemsa differential stain, and the average number of each cell type was determined after counting a total of 300 cells in 10-20 high-powered fields (×1,000) per slide. A total of 1 × 105 BAL cells were cytospun onto each slide to compensate for differences in cell retrieval.

Whole lung histological analysis. Whole lungs from Eo-/- and Eo+/+ mice were fully inflated with 10% formalin, dissected, and placed in fresh formalin for 24 h. Routine histological techniques were used to paraffin embed the entire lung, and 5-µm sections of whole lung were stained with hematoxylin and eosin. Inflammatory infiltrates and structural alterations were examined around small airways and adjacent blood vessels using light microscopy at a magnification of ×200.

ELISA analysis. Murine eotaxin/CCL11, interleukin (IL)-10, monocyte chemoattractant protein-1 (MCP-1)/CCL2, interferon-gamma (IFN-gamma ), IL-13, and C10/CCL6 chemokine were measured in 50-µl samples from whole lung homogenates (whole lung samples were initially homogenized in 1 ml of saline containing protease inhibitors) using a standardized sandwich ELISA technique previously described in detail (13). Each ELISA was screened to ensure antibody specificity and recombinant murine cytokines, and chemokines were used to generate the standard curves from which the concentrations present in the samples were derived. The limit of ELISA detection for each cytokine was consistently above 50 pg/ml. The cytokine and chemokine levels in each sample were normalized to total protein levels measured using the Bradford assay (Bio-Rad, Hercules, CA).

Statistical analysis. All results are expressed as means ± SE. Two separate experiments were performed. A Student's t-test was used to reveal statistical differences between the Eo-/- and Eo+/+ groups in both fungal asthma models. P < 0.05 was considered statistically significant.


    RESULTS
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ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Role of eotaxin in an acute model of A. fumigatus-induced allergic airway disease. A number of studies have addressed the effects of soluble Aspergillus antigens on the allergic inflammatory response within the murine lung (23, 24), and airway eosinophilia is a prominent feature of this response (20). In the present study, peak changes in whole lung and BAL levels of eotaxin/CCL11 were observed at 24 h after the soluble A. fumigatus antigen challenge (Fig. 1) in Eo+/+ mice. Although the increases in eotaxin/CCL1 levels in whole lung samples were significant at 3 and 24 h after the allergen challenge, these increases were less than a twofold increase above levels measured before the allergen challenge. No significant increases in BAL levels of eotaxin/CCL11 were noted at any time after the conidia challenge, rather, BAL levels of this chemokine were significantly lower at 48 and 72 h after the allergen challenge (compared with BAL levels before the allergen challenge; Fig. 1). Determination of methacholine-induced airway hyperresponsiveness at 24 and 48 h after the soluble allergen challenge revealed that methacholine-induced airway resistance in Eo-/- mice was lower at both time points compared with Eo+/+ mice, with a significant difference detected at 24 h after the allergen challenge (Fig. 2). Morphometric analysis of airway-associated eosinophilia revealed that significantly fewer eosinophils were present in Eo-/- mice at both times after the A. fumigatus challenge compared with the wild-type control group (Fig. 3A). In contrast, lymphocyte numbers (primarily T cells) were higher in the airways of Eo-/- mice at 24 and 48 h after the soluble allergen challenge, and this increase reached statistical significance at 48 h after the allergen challenge (Fig. 3B). One explanation for the increase in the numbers of T lymphocytes in the airways of Eo-/- mice may be related to the significant elevations in C10/CCL6 chemokine, a potent T cell chemoattractant (45), that were observed in the BAL of these mice at 48 h after the allergen challenge (Fig. 4). Histological examination of whole lung specimens from Eo-/- and Eo+/+ mice (Fig. 5) revealed that eosinophil recruitment to the airways was clearly impaired in Eo-/- mice at 24 h (Fig. 5D) and 48 h (Fig. 5F) compared with Eo+/+ mice (Fig. 5, C and E) at the same times. Together, it was clear that the recruitment of eosinophils in the context of acute fungus-induced allergic airway disease was markedly impaired, but this impairment did not fully inhibit the development of acute fungal asthma.


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Fig. 1.   Whole lung and bronchoalveolar lavage BAL eotaxin/CCL11 levels in Aspergillus fumigatus-sensitized wild-type (Eo+/+) mice at various times after soluble A. fumigatus (Asp) allergen challenge. Eotaxin/CCL11 levels were measured using specific ELISAs as described in MATERIALS AND METHODS. Data are expressed as means ± SE; n = 3 mice/group. * P <=  0.05, compared with levels measured before (t = 0) the allergen challenge.



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Fig. 2.   Airway hyperresponsiveness at 24 and 48 h after a soluble A. fumigatus allergen challenge in A. fumigatus-sensitized Eo+/+ mice and Eo-/- (lacking the eotaxin gene) mice. The baseline airway resistance in all groups was similar before the methacholine provocation, and these values (units = cmH2O · ml-1 · s-1) were as follows: Eo+/+ at 24 h, 1.5 ± 0.11; Eo+/+ at 48 h, 1.3 ± 0.5; Eo-/- at 24 h, 1.5 ± 0.14; Eo-/- at 48 h, 1.5 ± 0.5. Peak increases in airway resistance after the intravenous injection of 125 µg/ml of methacholine are shown. Values are expressed as means ± SE; n = 3 mice/group. *** P <=  0.001 compared with airway hyperresponsiveness measured in Eo+/+ mice at the same time after the soluble A. fumigatus allergen challenge.



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Fig. 3.   Temporal changes in eosinophil (A) and T lymphocyte (B) counts in BAL samples before and at 24 and 48 h after a soluble A. fumigatus allergen challenge in A. fumigatus-sensitized Eo+/+ and Eo-/- groups. BAL cells were dispersed onto microscope slides and were differentially stained with Wright-Giemsa stain. A minimum of 15 high-powered fields (HPF) or 300 cells was examined in each cytospin. A total of 1 × 105 BAL cells were cytospun onto each slide to compensate for differences in cell retrieval from each mouse.



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Fig. 4.   BAL levels of C10/CCL6 before (indicated by dashed line) and at 24 and 48 h after an A. fumigatus allergen challenge in A. fumigatus-sensitized Eo+/+ and Eo-/- mice. Immunoreactive C10/CCL6 levels were determined using specific ELISAs as described in MATERIALS AND METHODS. Values are expressed as means ± SE; n = 3 mice/group. P = 0.006 compared with C10/CCL6 levels measured in the Eo+/+ control group at the same day after the soluble A. fumigatus allergen challenge.



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Fig. 5.   Representative photomicrographs of hematoxylin and eosin-stained whole lung sections before and at 24 and 48 h after a soluble A. fumigatus allergen challenge in A. fumigatus-sensitized Eo+/+ and Eo-/- groups. Little peribronchial inflammation was observed in Eo+/+ (A) and Eo-/- (B) mice before the allergen challenge. At 24 h after the allergen challenge, a marked peribronchial accumulation of eosinophils and mononuclear cells was observed (C). In contrast, little peribronchial inflammation was observed in Eo-/- mice at this time (D). At 48 h after allergen, the peribronchial inflammation was very pronounced in Eo+/+ mice (E), whereas little peribronchial inflammation was apparent in Eo-/- mice at this time after allergen (F). Instead, there appeared to be pronounced perivascular accumulation of eosinophils and mononuclear cells. Original magnification was ×200 for each photomicrograph.

Role of eotaxin in a chronic model of A. fumigatus-induced allergic airway disease. We have recently described a model of Aspergillus-induced airway disease or chronic fungal asthma that persists for several weeks after the intratracheal introduction of A. fumigatus conidia into A. fumigatus-sensitized mice (19). We have also observed that many features of this disease, including peribronchial inflammation, hyperreactivity, and remodeling, are dependent on the action of various chemokines, including MCP-1/CCL2, major intrinsic protein (MIP)-1alpha /CCL3, regulated upon activation, normal T cell expressed, and, presumably, secreted (RANTES)/CCL5, and macrophage-derived chemokine/CCL22 (5-7). Eotaxin/CCL11 was measured by specific ELISA in whole lung and BAL samples from Eo+/+ mice before and at 3, 7, and 30 days after a conidia challenge (Fig. 6). In contrast to eotaxin/CCL11 levels in the acute fungal asthma model, no significant changes in eotaxin/CCL11 were observed at any time after the conidia challenge compared with levels before the conidia challenge. Measurement of airway hyperresponsiveness induced by exogenous methacholine challenge revealed similar increases in airway resistance at all times after the conidia challenge (Fig. 7). Histological examination at day 30 after conidia confirmed that eotaxin did not mediate the pulmonary disease associated with chronic fungal asthma. No differences in the degree of peribronchial inflammation or airway remodeling were observed between the two groups at this time (not shown). Differential counts of leukocytes in the BAL of Eo-/- and Eo+/+ mice at the day 30 time point revealed that macrophages were present in significantly higher numbers in Eo-/- mice compared with Eo+/+ mice, but all other leukocyte subtypes were similar in both groups (Fig. 8). Some notable differences were observed in whole lung cytokine and chemokine levels in both groups of mice at day 30 after the conidia challenge (Fig. 9A). Whole lung IL-10 and MCP-1/CCL2 levels were significantly lower, whereas IFN-gamma levels were fourfold lower in Eo-/- mice than levels of the same cytokines and chemokines measured in whole lung samples from Eo+/+ mice (Fig. 9B). However, prominent levels of IL-13 and C10/CCL6 were measured in whole lung samples from both groups at this time. Both IL-13 and C10/CCL6 have significant roles in the maintenance of chronic fungal asthma (3, 4).


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Fig. 6.   Whole lung and BAL eotaxin/CCL11 levels in A. fumigatus-sensitized Eo+/+ mice at various times after A. fumigatus conidia challenge. Eotaxin/CCL11 levels were measured using specific ELISAs as described in MATERIALS AND METHODS. Data are expressed as means ± SE; n = 5 mice/group.



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Fig. 7.   Airway hyperresponsiveness at days 3, 7, and 30 after an A. fumigatus conidia challenge in A. fumigatus-sensitized Eo+/+ and Eo-/- mice. The baseline airway resistance in all groups was similar before the methacholine provocation, and these values (units = cmH2O · ml-1 · s-1) were as follows: Eo+/+ at day 3, 1.5 ± 0.4; Eo+/+ at day 7, 1.2 ± 0.1; Eo+/+ at day 30, 1.7 ± 0.4; Eo-/- at day 3, 1.4 ± 0.01; Eo-/- at day 7, 1.3 ± 0.2; Eo-/- at day 30, 2.1 ± 0.7. Peak increases in airway resistance after the intravenous injection of 125 µg/ml of methacholine are shown. Values are expressed as means ± SE; n = 5 mice/group.



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Fig. 8.   Differential leukocyte counts in BAL samples at day 30 after an A. fumigatus conidia challenge in A. fumigatus-sensitized Eo+/+ and Eo-/- groups. BAL cells were dispersed onto microscope slides and were differentially stained with Wright-Giemsa stain. A minimum of 15 HPF or 300 cells was examined in each cytospin. A total of 1 × 105 BAL cells were cytospun onto each slide to compensate for differences in cell retrieval from each mouse.



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Fig. 9.   Whole lung interleukin (IL)-10, monocyte chemoattractant protein-1 (MCP-1)/CCL2, and interferon-gamma (IFN-gamma ) (A) and IL-13 and C10/CCL6 (B) levels in A. fumigatus-sensitized Eo+/+ and Eo-/- mice at day 30 after an A. fumigatus conidia challenge. Cytokine and chemokine levels were measured using specific ELISAs as described in MATERIALS AND METHODS. Data are expressed as means ± SE; n = 5 mice/group.


    DISCUSSION
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ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

Considering the important role that eotaxin/CCL11 exerts on the early recruitment of eosinophils during allergic and asthmatic responses (39) and its putative role in attracting Th2 cells (42), this chemokine is an attractive target in these diseases (9). The peribronchial infiltration of eosinophils is mediated in part by allergen-specific Th2 cells, and supernatants from Th2 cells induced eotaxin/CCL11, RANTES/CCL5, and lung eosinophilia in naive mice lung eosinophilia when administered intranasally (28). Other experiments showed that eotaxin/CCL11 was a major link between allergen-specific T cell activation and the recruitment of eosinophils into the airways (34). From the present study, it is apparent that eotaxin/CCL11 regulates eosinophil recruitment at early times after a soluble A. fumigatus allergen challenge into A. fumigatus-sensitized mice, but its role in eosinophil recruitment appears to be limited during more protracted responses (i.e., chronic fungal asthma) to A. fumigatus spores. This finding is consistent with previous studies in eotaxin-deficient mice in which eosinophil recruitment was not (47), or only partially (39), inhibited after ovalbumin sensitization and challenge. Thus the genetic targeting of eotaxin/CCL11 significantly inhibits, but does not ablate, eosinophil recruitment in the context of acute fungus-induced asthma.

The present study also highlighted that the lack of eotaxin/CCL11 did not impact the recruitment of T lymphocytes into the airways of A. fumigatus-challenged mice. It appears that the development of allergic disease due to Aspergillus is initially dependent on the movement of eosinophils into the airways of allergic mice, but other chemokines or factors and T cells then appear to regulate airway hyperresponsiveness and the other features of fungal asthma. Two prime candidates are C10/CCL6 and the pleiotropic cytokine IL-13. C10/CCL6 has major effects on the recruitment of eosinophils and T cells during acute fungal asthma (20), whereas IL-13 is a major effector in the maintenance of all features of chronic fungal asthma (3, 4). In addition, we have observed that Th2 cells have a prominent role in the development and maintenance of chronic fungal asthma, and disease resolution only appears in the context of this disease when Th2 cell recruitment or activation is inhibited (3, 4).

The observation that experimental fungal asthma proceeds in the absence of eotaxin/CCL11 presumably reflects the fact that various chemokines, including MIP-1alpha /CCL3, C10/CCL6, RANTES/CCL5, and other eotaxins including eotaxin-2/CCL24 (14) and eotaxin-3/CCL26 (43), have chemotactic effects on eosinophils and Th2 cells during allergic and asthmatic responses (31). In addition, it has been shown in a number of studies that distinct functional groups of chemokines cooperate and coordinate the pulmonary inflammatory response during experimental allergy and asthma (8, 16, 18). Although this possibility was not explored in the context of the murine models described herein or elsewhere, more recent clinical data suggest that eotaxin-3/CCL26 rather than eotaxin/CCL11 or eotaxin-2/CCL24 sustains eosinophil recruitment to asthmatic airways in the later stages after allergen challenge (1).

In conclusion, the present study demonstrates that eotaxin/CCL11 has a major role in the recruitment of eosinophils into the airways during early allergic responses to A. fumigatus, but its role is limited during more chronic responses to this fungus. In addition, the lack of eotaxin/CCL11 did not affect the recruitment of T cells to the airways in either model of Aspergillus-induced disease. Considering these findings, it is probable that the targeting of eotaxin/CCL11 during A. fumigatus allergen or conidia challenge may be of limited utility. However, these findings do not rule out the therapeutic benefit that may be obtained with a specific CCR3 antagonist (2, 41) that would block the action of all eotaxins and other CCR3 agonists.


    ACKNOWLEDGEMENTS

This work was supported, in part, by funding from the American Lung Association.


    FOOTNOTES

Address for reprint requests and other correspondence: C. M. Hogaboam, Dept. of Pathology, Univ. of Michigan Medical School, 1301 Catherine Road, Ann Arbor, MI 48109-0602 (E-mail: hogaboam{at}med.umich.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.

First published February 8, 2002;10.1152/ajplung.00341.2001

Received 27 August 2001; accepted in final form 5 February 2002.


    REFERENCES
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

1.   Berkman, N, Ohnona S, Chung FK, and Breuer R. Eotaxin-3 but not eotaxin gene expression is upregulated in asthmatics 24 hours after allergen challenge. Am J Respir Cell Mol Biol 24: 682-687, 2001[Abstract/Free Full Text].

2.   Bertrand, CP, and Ponath PD. CCR3 blockade as a new therapy for asthma. Expert Opin Investig Drugs 9: 43-52, 2000[ISI][Medline].

3.   Blease, K, Jakubzick C, Westwick J, Lukacs N, Kunkel SL, and Hogaboam CM. Therapeutic effect of IL-13 immunoneutralization during chronic experimental fungal asthma. J Immunol 166: 5219-5224, 2001[Abstract/Free Full Text].

4.   Blease, K, Jakubzick C, Schuh JM, Joshi BH, Puri RK, and Hogaboam CM. IL-13 fusion cytotoxin ameliorates chronic fungal-induced allergic airway disease in mice. J Immunol 167: 6583-6592, 2001[Abstract/Free Full Text].

5.   Blease, K, Mehrad B, Lukacs NW, Kunkel SL, Standiford TJ, and Hogaboam CM. Antifungal and airway remodeling roles for murine monocyte chemoattractant protein-1/CCL2 during pulmonary exposure to Aspergillus fumigatus conidia. J Immunol 166: 1832-1842, 2001[Abstract/Free Full Text].

6.   Blease, K, Mehrad B, Standiford TJ, Lukacs NW, Gosling J, Boring L, Charo IF, Kunkel SL, and Hogaboam CM. Enhanced pulmonary allergic responses to Aspergillus in CCR2-/- mice. J Immunol 165: 2603-2611, 2000[Abstract/Free Full Text].

7.   Blease, K, Mehrad B, Standiford TJ, Lukacs NW, Kunkel SL, Chensue SW, Lu B, Gerard CJ, and Hogaboam CM. Airway remodeling is absent in CCR1-/- mice during chronic fungal allergic airway disease. J Immunol 165: 1564-1572, 2000[Abstract/Free Full Text].

8.   Campbell, EM, Kunkel SL, Strieter RM, and Lukacs NW. Temporal role of chemokines in a murine model of cockroach allergen-induced airway hyperreactivity and eosinophilia. J Immunol 161: 7047-7053, 1998[Abstract/Free Full Text].

9.   Corrigan, C. The eotaxins in asthma and allergic inflammation: implications for therapy. Curr Opin Investig Drugs 1: 321-328, 2000[Medline].

10.   Crump, MP, Rajarathnam K, Kim KS, Clark-Lewis I, and Sykes BD. Solution structure of eotaxin, a chemokine that selectively recruits eosinophils in allergic inflammation. J Biol Chem 273: 22471-22479, 1998[Abstract/Free Full Text].

11.   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].

12.   Elsner, J, Hochstetter R, Kimmig D, and Kapp A. Human eotaxin represents a potent activator of the respiratory burst of human eosinophils. Eur J Immunol 26: 1919-1925, 1996[ISI][Medline].

13.   Evanoff, H, Burdick MD, Moore SA, Kunkel SL, and Strieter RM. A sensitive ELISA for the detection of human monocyte chemoattractant protein-1 (MCP-1). Immunol Invest 21: 39-49, 1992[ISI][Medline].

14.   Forssmann, U, Uguccioni M, Loetscher P, Dahinden CA, Langen H, Thelen M, and Baggiolini M. Eotaxin-2, a novel CC chemokine that is selective for the chemokine receptor CCR3, and acts like eotaxin on human eosinophil and basophil leukocytes. J Exp Med 185: 2171-2176, 1997[Abstract/Free Full Text].

15.   Gao, JL, Sen AI, Kitaura M, Yoshie O, Rothenberg ME, Murphy PM, and Luster AD. Identification of a mouse eosinophil receptor for the C-C chemokine eotaxin. Biochem Biophys Res Commun 223: 679-684, 1996[ISI][Medline].

16.   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[Abstract/Free Full Text].

17.   Griffiths-Johnson, DA, Collins PD, Rossi AG, Jose PJ, and Williams TJ. The chemokine, eotaxin, activates guinea-pig eosinophils in vitro and causes their accumulation in vivo. Biochem Biophys Res Commun 197: 1167-1172, 1993[ISI][Medline].

18.   Gutierrez-Ramos, JC, Lloyd C, Kapsenberg ML, Gonzalo JA, and Coyle AJ. Non-redundant functional groups of chemokines operate in a coordinate manner during the inflammatory response in the lung. Immunol Rev 177: 31-42, 2000[ISI][Medline].

19.   Hogaboam, CM, Blease K, Mehrad B, Steinhauser ML, Standiford TJ, Kunkel SL, and Lukacs NW. Chronic airway hyperreactivity, goblet cell hyperplasia, and peribronchial fibrosis during allergic airway disease induced by Aspergillus fumigatus. Am J Pathol 156: 723-732, 2000[Abstract/Free Full Text].

20.   Hogaboam, CM, Gallinat CS, Taub DD, Strieter RM, Kunkel SL, and Lukacs NW. Immunomodulatory role of C10 chemokine in a murine model of allergic bronchopulmonary aspergillosis. J Immunol 162: 6071-6079, 1999[Abstract/Free Full Text].

21.   Jose, PJ, Griffiths-Johnson DA, Walsh DT, Moqbel R, Totty NF, Truong O, Hsuan JJ, and Williams TJ. Eotaxin: a potent eosinophil chemoattractant cytokine detected in a guinea-pig model of allergic airway inflammation. J Exp Med 179: 881-887, 1994[Abstract].

22.   Kauffman, HF, Tomee JF, van der Werf TS, de Monchy JG, and Koeter GK. Review of fungus-induced asthmatic reactions. Am J Respir Crit Care Med 151: 2109-2115, 1995[Abstract].

23.   Kurup, VP, Mauze S, Choi H, Seymour BW, and Coffman RL. A murine model of allergic bronchopulmonary aspergillosis with elevated eosinophils and IgE. J Immunol 148: 3783-3788, 1992[Abstract/Free Full Text].

24.   Kurup, VP, Seymour BWP, Choi H, and Coffman RL. Particulate Aspergillus fumigatus antigens elicit a Th2 response in BALB/c mice. J Allergy Clin Immunol 93: 1013-1020, 1994[ISI][Medline].

25.   Kurup, VP, Shen HD, and Banerjee B. Respiratory fungal allergy. Microbes Infect 2: 1101-1110, 2000[ISI][Medline].

26.   Lamkhioued, B, Renzi PM, Abi-Younes S, Garcia-Zepeda EA, Allakhverdi Z, Ghaffar O, Rothenberg MD, Luster AD, and Hamid Q. Increased expression of eotaxin in bronchoalveolar lavage and airways of asthmatics contributes to the chemotaxis of eosinophils to the site of inflammation. J Immunol 159: 4593-4601, 1997[Abstract].

27.   Latge, JP. Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev 12: 310-350, 1999[Abstract/Free Full Text].

28.   Li, L, Xia Y, Nguyen A, Lai YH, Feng L, Mosmann TR, and Lo D. Effects of Th2 cytokines on chemokine expression in the lung: IL-13 potently induces eotaxin expression by airway epithelial cells. J Immunol 162: 2477-2487, 1999[Abstract/Free Full Text].

29.   Lilly, CM, Nakamura H, Belostotsky OI, Haley KJ, Garcia-Zepeda EA, Luster AD, and Israel E. Eotaxin expression after segmental allergen challenge in subjects with atopic asthma. Am J Respir Crit Care Med 163: 1669-1675, 2001[Abstract/Free Full Text].

30.   Lloyd, CM, Delaney T, Nguyen T, Tian J, Martinez AC, Coyle AJ, and Gutierrez-Ramos JC. CC chemokine receptor (CCR)3/eotaxin is followed by CCR4/monocyte-derived chemokine in mediating pulmonary T helper lymphocyte type 2 recruitment after serial antigen challenge in vivo. J Exp Med 191: 265-274, 2000[Abstract/Free Full Text].

31.   Lukacs, NW, Oliveira SH, and Hogaboam CM. Chemokines and asthma: redundancy of function or a coordinated effort? J Clin Invest 104: 995-999, 1999[Free Full Text].

32.   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].

33.   Lukacs, NW, Strieter RM, Chensue SW, and Kunkel SL. TNF-alpha mediates recruitment of neutrophils and eosinophils during allergic airway inflammation. J Immunol 154: 5411-5417, 1993[Abstract/Free Full Text].

34.   MacLean, JA, Ownbey R, and Luster AD. T cell-dependent regulation of eotaxin in antigen-induced pulmonary eosinophila. J Exp Med 184: 1461-1469, 1996[Abstract].

35.   Mould, AW, Ramsay AJ, Matthaei KI, Young IG, Rothenberg ME, and Foster PS. The effect of IL-5 and eotaxin expression in the lung on eosinophil trafficking and degranulation and the induction of bronchial hyperreactivity. J Immunol 164: 2142-2150, 2000[Abstract/Free Full Text].

36.   Ponath, PD, Qin S, Ringler DJ, Clark-Lewis I, Wang J, Kassam N, Smith H, Shi X, Gonzalo JA, Newman W, Gutierrez-Ramos JC, and Mackay CR. Cloning of the human eosinophil chemoattractant, eotaxin. Expression, receptor binding, and functional properties suggest a mechanism for the selective recruitment of eosinophils. J Clin Invest 97: 604-612, 1996[Abstract/Free Full Text].

37.   Rankin, SM, Conroy DM, and Williams TJ. Eotaxin and eosinophil recruitment: implications for human disease. Mol Med Today 6: 20-27, 2000[ISI][Medline].

38.   Rothenberg, ME, Luster AD, and Leder P. Murine eotaxin: an eosinophil chemoattractant inducible in endothelial cells and in interleukin 4-induced tumor suppression. Proc Natl Acad Sci USA 92: 8960-8964, 1995[Abstract].

39.   Rothenberg, ME, MacLean JA, Pearlman E, Luster AD, and Leder P. Targeted disruption of the chemokine eotaxin partially reduces antigen-induced tissue eosinophilia. J Exp Med 185: 785-790, 1997[Abstract/Free Full Text].

40.   Rothenberg, ME, Ownbey R, Mehlhop PD, Loiselle PM, van de Rijn M, Bonventre JV, Oettgen HC, Leder P, and Luster AD. Eotaxin triggers eosinophil-selective chemotaxis and calcium flux via a distinct receptor and induces pulmonary eosinophilia in the presence of interleukin 5 in mice. Mol Med 2: 334-348, 1996[ISI][Medline].

41.   Sabroe, I, Peck MJ, Van Keulen BJ, Jorritsma A, Simmons G, Clapham PR, Williams TJ, and Pease JE. A small molecule antagonist of chemokine receptors CCR1 and CCR3. Potent inhibition of eosinophil function and CCR3-mediated HIV-1 entry. J Biol Chem 275: 25985-25992, 2000[Abstract/Free Full Text].

42.   Sallusto, F, Mackay CR, and Lanzavecchia A. Selective expression of the eotaxin receptor CCR3 by human T helper 2 cells. Science 277: 2005-2007, 1997[Abstract/Free Full Text].

43.   Shinkai, A, Yoshisue H, Koike M, Shoji E, Nakagawa S, Saito A, Takeda T, Imabeppu S, Kato Y, Hanai N, Anazawa H, Kuga T, and Nishi T. A novel human CC chemokine, eotaxin-3, which is expressed in IL-4-stimulated vascular endothelial cells, exhibits potent activity toward eosinophils. J Immunol 163: 1602-1610, 1999[Abstract/Free Full Text].

44.   Taha, RA, Minshall EM, Miotto D, Shimbara A, Luster A, Hogg JC, and Hamid QA. Eotaxin and monocyte chemotactic protein-4 mRNA expression in small airways of asthmatic and nonasthmatic individuals. J Allergy Clin Immunol 103: 476-483, 1999[ISI][Medline].

45.   Wang, W, Bacon KB, Oldham ER, and Schall TJ. Molecular cloning and functional characterization of human MIP-1 delta, a new C-C chemokine related to mouse CCF-18 and C10. J Clin Immunol 18: 214-222, 1998[ISI][Medline].

46.   Yamada, H, Yamaguchi M, Yamamoto K, Nakajima T, Hirai K, Morita Y, and Sano Y. Eotaxin in induced sputum of asthmatics: relationship with eosinophils and eosinophil cationic protein in sputum. Allergy 55: 392-397, 2000[ISI][Medline].

47.   Yang, Y, Loy J, Ryseck RP, Carrasco D, and Bravo R. Antigen-induced eosinophilic lung inflammation develops in mice deficient in chemokine eotaxin. Blood 92: 3912-3923, 1998[Abstract/Free Full Text].

48.   Ying, S, Meng Q, Zeibecoglou K, Robinson DS, Macfarlane A, Humbert M, and Kay AB. Eosinophil chemotactic chemokines [eotaxin, eotaxin-2, RANTES, monocyte chemoattractant protein-3 (MCP-3), and MCP-4] and C-C chemokine receptor 3 expression in bronchial biopsies from atopic and nonatopic (Intrinsic) asthmatics. J Immunol 163: 6321-6329, 1999[Abstract/Free Full Text].

49.   Ying, S, Robinson DS, Meng Q, Rottman J, Kennedy R, Ringler DJ, Mackay CR, Daugherty BL, Springer MS, Durham SR, Williams TJ, and Kay AB. Enhanced expression of eotaxin and CCR3 mRNA and protein in atopic asthma. Association with airway hyperresponsiveness and predominant co-localization of eotaxin mRNA to bronchial epithelial and endothelial cells. Eur J Immunol 27: 3507-3516, 1997[ISI][Medline].


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