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Increases in inflammatory cell progenitors, particularly eosinophil/basophil colony-forming cells (Eo/ B-CFU), occur in peripheral blood after allergen provocation. The role of bone marrow (BM) in these reactions is unclear. We examined the effect of allergen challenge on human bone marrow progenitor cell growth. Fifteen asthmatic subjects, eight dual responders (DR) and seven isolated early responders (IER), were challenged with inhaled allergen. BM aspirates were taken before and 24 h after challenge and progenitors were enumerated by a colony-forming assay. Eo/B-CFU numbers increased in both groups after allergen challenge (p < 0.0001). For DR, the increases were significant for BM incubated with optimal GMCSF and IL-5, but not with IL-3. For IER, the increases were significant for all three cytokines tested. At a suboptimal concentration of IL-5, there was a significant increase in the number of Eo/B-CFU after allergen in the DR, from 5.25 ± 1.2 to 9.68 ± 2.1 per 2.5 × 105 cells plated (p < 0.01), which was not demonstrated in the IER (p = 0.94). The responses at this concentration of IL-5 were different between groups (p < 0.05). These results demonstrate that inhaled allergen increases BM Eo/B-CFU, and that the bone marrow of dual responders is more responsive to IL-5 after allergen.
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INTRODUCTION |
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ABSTRACT
INTRODUCTION
METHODS
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Asthma is a disease characterized by bronchoconstriction, airway hyperresponsiveness, and airway inflammation. Inhalation of allergen by sensitized subjects is an important cause of asthma, and is characterized by biphasic responses known as the early- and late-phase asthmatic responses. Late asthmatic responses (LAR) are associated with transient increases in airway hyperresponsiveness (1), usually lasting several days, and increases in the numbers of activated eosinophils and metachromatic cells in the airways (2); however, subjects developing isolated early asthmatic responses also develop airway hyperresponsiveness, but to a much lesser extent than those patients developing late responses (3).
The predominant cell infiltrating the airways during the late response is the eosinophil. These cells are selectively increased in sputum (4) and bronchoalveolar lavage fluid (BAL) (5), in association with the late response. The eosinophils are also in an activated state in asthma, as indicated by elevated levels of eosinophil cationic protein (ECP) in BAL fluid (6), and enhanced immunostaining with the EG2 monoclonal antibody, which recognizes the cleaved and activated form of ECP.
We have previously provided evidence that an important aspect of allergic inflammatory responses is the induction of increases in inflammatory cell progenitors, which contribute to disease through the continued production of inflammatory effector cells (7, 8). Higher numbers of both circulating eosinophil/basophil colony-forming units (Eo/B-CFU) and CD34+ hemopoietic progenitor cells are demonstrable in the blood of atopic subjects when compared with normal subjects (7, 9). In addition, the numbers of Eo/B-CFU in the bloodstream of asthmatic subjects at the time of an acute exacerbation is significantly higher than those measured after resolution of the exacerbation (10). Also, increased Eo/B-CFU can be stimulated from the peripheral blood of atopic subjects in the presence of conditioned media from atopic nasal, mucosal, and epithelial cells (11), suggesting the production of colony-stimulating activity from these cells. In vivo studies in atopic subjects have shown that there are fluctuating numbers of circulating Eo/B-CFU during seasonal exposure to allergen (12) and significantly higher numbers 24 h after allergen inhalation (13). Finally, in dogs with allergen-induced airway hyperresponsiveness and airway inflammation, bone marrow granulocyte-macrophage colony-forming units (GM-CFU) are significantly increased after allergen challenge (14).
Although such studies suggest that inflammatory cell progenitors in the blood arise from the bone marrow, this has not been shown in humans. The purposes of this study were fourfold: (1) to determine whether allergen inhalation increases the production of bone marrow inflammatory cell progenitors in sensitized subjects with mild asthma; (2) to evaluate whether, in subjects who develop both an early and a late asthmatic response (dual responders) and in subjects who develop isolated early responses, the differential physiologic and inflammatory responses to allergen are reflected in different bone marrow responses; (3) to establish whether the changes in bone marrow progenitors are selective for Eo/B colonies; (4) to determine whether there were differences in the bone marrow's responsiveness after allergen to optimal and suboptimal concentration of hemopoietic cytokines.
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INTRODUCTION
METHODS
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DISCUSSION
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Subjects
Fifteen patients with mild asthma (eight dual responders and seven
isolated early responders) were studied (Table 1). Subjects were classified as dual responders if they developed both early- and late-phase
asthmatic responses as defined by a greater than 15% drop in FEV1
from baseline, or as isolated early responders if they developed only
an early fall in FEV1 greater than 15% from baseline. The study was
approved by the Research Advisory Group at McMaster University
Medical Centre, and all subjects provided their written informed consent prior to entering the study. Subjects were atopic, as indicated by
one or more positive wheal-and-flare responses to skin prick tests. All
subjects were nonsmokers and none had experienced a respiratory infection during the 4 wk prior to the study. Asthmatic subjects were
stable at the time of study, requiring only intermittent use of inhaled 
2-agonists, with baseline FEV1 values > 70% predicted.
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Study Design
Subjects attended the laboratory on three occasions. At the initial visit a full medical history was taken and skin prick tests were performed. In addition, spirometry and methacholine inhalation challenge were performed followed by induction of sputum. Within 1 wk subjects returned to the laboratory to undergo an allergen-challenge procedure. Prior to allergen challenge, a bone marrow aspirate and blood sample were taken to determine baseline measurements. At 5 h after allergen inhalation a second blood sample was withdrawn. The third visit to the laboratory occurred 24 h after allergen inhalation. At this visit, postallergen bone marrow aspirate and blood samples were obtained and spirometry and a methacholine inhalation challenge were performed, followed by a sputum induction procedure to evaluate the airway cellular responses.
Methacholine Inhalation Challenge
Methacholine inhalation was performed as described by Cockcroft (15). Subjects inhaled normal saline and then doubling concentrations of methacholine phosphate from a Wright nebulizer for 2 min. Increasing concentrations of methacholine were administered until the FEV1 decreased by > 20% of the baseline value. Results were expressed as the geometric mean provocative concentration causing a 20% decrease in FEV1 (PC20). For the skin prick titration test, the allergen to be used in the allergen inhalation challenge was administered in doubling dilutions in duplicate.
Allergen Inhalation Challenge
Allergen inhalation challenge was performed as described by O'Byrne (16). The allergen producing the largest skin wheal diameter was diluted in normal saline. The concentration of allergen extract for inhalation was determined from a formula described by Cockcroft and colleagues (17) using results from the skin test and the methacholine PC20. The starting concentration of allergen was chosen to be three doubling doses below that predicted to cause a 20% fall in FEV1. Doubling concentrations of allergen were inhaled at 10-min intervals until a decrease of 15% or more occurred in the FEV1 from baseline. Measurements of FEV1 were performed at 10, 20, 30, 45, 60, 90, and 180 min, and then every hour for 7 h after the final inhalation. The allergen-induced early response was determined as the maximal decrease in FEV1 between 0 and 2 h after allergen, and the late response was determined as the maximal decrease between 3 and 7 h after allergen inhalation.
Blood Samples
Venous blood samples were obtained from each subject before and at 5 and 24 h after allergen inhalation. Samples were collected in tubes treated with ethylenediaminetetraacetic acid (EDTA) for total and differential WBC counts. Total cell counts were performed using a neubauer hemocytometer, and differential cell counts were made from blood smears stained by Diff-Quik (American Scientific Products, McGaw Park, IL). Differential cell counts were performed by one investigator in a blinded fashion and the mean of two slides was obtained (300 cells counted per slide). Cells were classified using standard morphologic criteria. Results were expressed as absolute counts (109 cells/L).
Bone Marrow Aspirate and Culture
Bone marrow aspirates were obtained from the iliac crest using a
bone marrow aspiration needle (16 × 2''; Sherwood Medical, St.
Louis, MO). Three milliliters of bone marrow were aspirated into a
10-ml syringe containing 1 ml sterile heparin (1,000 U/ml) (Leo Laboratories, ON, Canada) and semisolid methylcellulose cultures of low
density nonadherent mononuclear cells were performed. Briefly, heparinized bone marrow was diluted to 50 ml with McCoy's 5A medium (GIBCO, Grand Island, NY) and separated over 65% percoll (Pharmacia, Uppsala, Sweden). The interface mononuclear-rich cell fraction was washed in McCoy's 5A medium and then incubated in McCoy's 5A medium supplemented with 15% fetal calf serum (FCS)
(GIBCO), 1% penicillin/streptomycin (GIBCO), and 5 × 10
5 M
2-mercaptoethanol (final concentration) (Sigma Chemical, St. Louis,
MO) for 2 h in plastic flasks at 37° C and 5% CO2. Nonadherent mononuclear cells (NAMC) containing progenitor cells and lymphocytes were then cultured (2.5 × 10 5 cells per 35 × 10 mm tissue culture
dish; Falcon Plastics, Oxnard, CA), in duplicate, in supplemented Iscove's modified Dulbecco's medium (GIBCO) with 1% penicillin/
streptomycin and 5 × 105 M 2-mercaptoethanol (final concentration), 0.9% methylcellulose (Sigma Chemical), and 20% FCS, either
alone or in the presence of one of the following growth factors: recombinant human IL-3 (1 or 0.1 ng/ml; Pharmingen, San Diego, CA), recombinant human GM-CSF (10 or 1 ng/ml; Pharmingen), or recombinant human IL-5 (1 or 0.1 ng/ml; Pharmingen). Cultures were
incubated for 14 d at 37° C and 5% CO2 after which colonies were
identified as either Eo/B-CFU or GM-CFU according to previously
described criteria (18) and expressed as colony-forming units (CFU)
per 2.5 × 105 NAMC plated.
Sputum Analysis
Sputum was induced and processed according to the method of Popov and colleagues (19). Subjects inhaled 3, 4, and then 5% saline for 10 min each until an adequate sample was obtained or if the FEV1 dropped 20% from baseline. Cell plugs were selected from the sample and processed using 0.1% dithiothreitol (Sputolysin; Calbiochem, San Diego, CA) and Dulbecco's phosphate-buffered saline (GIBCO). Cytospins were prepared on glass slides, and differential counts were performed in a blinded fashion on Diff-Quik-stained slides. Mean counts from duplicate slides were obtained (500 cells counted per slide) and expressed as percentages. When possible, cytospins were also prepared on APTEX-coated slides and fixed in periodate-lysine-paraformaldehyde (PLP) for immunocytochemical staining for eosinophil cationic protein (ECP) using a monoclonal antibody against cleaved ECP (EG2) (Kabi Pharmacia) (20). Results are expressed as percentages of 500 cells counted under light microscopy.
Statistical Analysis
Airway hyperresponsiveness. Methacholine PC20 values were log10 transformed prior to analysis. Preallergen versus postallergen comparisons were performed for each group using Student's paired t tests. The preallergen versus postallergen change in logged PC20 was compared between groups using Student's independent t test.
Sputum. Preallergen versus postallergen changes in induced sputum cell percentages were assessed for each group using Student's paired t tests. Differences in preallergen versus postallergen changes in cell percentages between each group were assessed using Student's nonpaired t tests. Regression analysis was used to detect significant relationships between induced sputum and changes in airway function (21).
Blood. Differences in blood differentials before and 5 and 24 h after allergen were assessed for each group using Student's paired t test.
Bone marrow colonies. Differences in bone marrow progenitor colonies were investigated using a mixed model analysis of variance (nonrepeated factor: early versus dual; repeated factors: preallergen versus postallergen, GM-CSF versus IL-3 versus IL-5, low versus high concentration) (22). Preallergen versus postallergen colony growth with each cytokine at each concentration comparisons were also performed as planned comparisons, using Student's paired t tests. Differences in the preallergen versus postallergen change in colony numbers between the two groups were analyzed using Student's nonpaired t tests. Statistical significance was assumed at p < 0.05.
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RESULTS |
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Bronchoconstrictor Responses
The mean maximal percent fall in FEV1 during the early asthmatic response was 25.9 ± 2.3% in the dual responders and 24.2 ± 2.1% in the isolated early responders (Table ). The maximal percent fall in FEV1 during the late asthmatic response was 22.6 ± 2.8% in the dual responders and 6.1 ± 2.7% in the isolated early responders (Table ).
Airway Hyperresponsiveness
Methacholine airway hyperresponsiveness developed in the dual responders but not in the isolated early responders 24 h after inhaled allergen. The geometric mean methacholine PC20 values in the dual responders fell from 1.66 mg/ml (%SEM, 1.28) before to 0.52 mg/ml (%SEM, 1.32) after allergen (p < 0.001), whereas in the isolated early responders the values were 0.98 mg/ml (%SEM, 1.61) before and 1.14 mg/ml (%SEM, 1.77) after allergen (p = 0.72) (Figure 1). In addition, there was a significant difference between the two groups in the allergen-induced shift in logged PC20 values (p < 0.001) (Figure 1).
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Airway Inflammation
The proportion of eosinophils in sputum after allergen challenge increased significantly in the dual responder group, from 3.4 ± 0.8% before to 32.7 ± 9.9% after allergen (p < 0.05) (Table 2). A smaller, not significant increase was seen in the isolated early responder group, where the values were 6.9 ± 2.1% before and 18.9 ± 9.7% after allergen (p = 0.28) (Table ). There were no significant differences in the increases in eosinophil numbers between the two groups (p = 0.23). Also, there were nonsignificant increases in the proportion of EG2-positive cells after allergen inhalation in both groups: 2.5 ± 1.1% before and 19.0 ± 10.4% after allergen in the dual responders (p = 0.15), and 2.1 ± 1.0% before and 11.2 ± 7.3% after in the isolated early responders (p = 0.28) (Table ). There were no significant correlations between changes in either sputum eosinophils or sputum EG2+ cells, with changes in log PC20 values (p > 0.05). The only other change in sputum inflammatory cells was a significant reduction in the percent macrophages in the dual responders (p = 0.007) (Table ).
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Blood Eosinophils
Blood eosinophils in the dual responder group fell from 0.40 ± 0.11 × 109/L before allergen to 0.22 ± 0.05 × 109/L 5 h after allergen (p < 0.05) (Table ). Subsequently, blood eosinophils increased from 0.22 ± 0.05 × 109/L to 0.32 ± 0.05 × 109/L between 5 and 24 h after allergen (p < 0.05) (Table ). There were no significant differences in blood eosinophils after allergen inhalation in the isolated early responders.
Bone Marrow Progenitors
The number of bone marrow Eo/B-CFU increased in both groups after allergen challenge (p < 0.0001) (Table 3). In the dual responders, the increases were significant for bone marrow grown in the presence of GM-CSF (10 ng/ml) and IL-5 (1 ng/ml), but not IL-3 (1 ng/ml) (Table ). In the isolated early responders, the increases were significant for the high concentrations of all three cytokines tested (Table ). At these concentrations, there were no significant differences in the magnitude of the Eo/B-CFU response between groups (p > 0.05). However, at the lower concentrations of IL-5 (0.1 ng/ ml), there was a significant increase in the number of Eo/B-CFU after allergen in the dual responders, from 5.3 ± 1.2 before to 9.7 ± 2.1 per 2.5 × 105 cells plated after allergen (p < 0.01), but not in the isolated early responders where the Eo/ B-CFU were 7.8 ± 3.0 before and 7.8 ± 2.6 per 2.5 × 105 cells plated after allergen (p = 0.94) (Figure 2 and Table ). There was a significant difference in the preallergen to postallergen change in Eo/B-CFU between the two groups at this concentration of IL-5 (p < 0.05). There were no significant increases in Eo/B-CFU after allergen inhalation in the presence of lower concentrations of IL-3 (0.1 ng/ml) or GM-CSF (1 ng/ml) in either subject group (Table ).
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p < 0.05.
There were no changes in the number of GM-CFU grown from the postallergen bone marrow in either group when incubated in the presence of IL-3 or IL-5 (Table ). There was, however, a significant increase in the number of GM-CFU grown from the postallergen bone marrow compared with the preallergen bone marrow when the cells were incubated with GM-CSF (10 ng/ml) in the dual responders (Table ). There was a tendency towards an increase in the number of GM-CFU after allergen seen in the isolated early responder group (Table ). There were no significant differences in the magnitude of the GM-CFU response between the two groups with any of the cytokines tested (Table ). Finally, there were no significant changes in the number of GM-CFU in the postallergen bone marrow samples from either group, when incubated with IL-3, GM-CSF, or IL-5 at suboptimal cytokine concentrations (Table ).
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This study has demonstrated an increase in the cytokine-induced production of bone marrow inflammatory cell progenitors associated with the development of airway hyperresponsiveness and airway inflammation after allergen inhalation in sensitized asthmatic subjects. In addition, only the dual responders demonstrated measurable methacholine airway hyperresponsiveness after allergen, and increases in sputum eosinophils were larger and significant in the dual responders. This result is consistent with other studies that have contrasted the airway inflammatory response between isolated early and dual responders (23).
Increases in bone marrow Eo/B-CFU were demonstrated in both subject groups after allergen challenge, when optimal concentrations (as determined in preliminary studies) of IL-3, IL-5, and GM-CSF were used. There was, however, a significant difference between the two groups when the cells were incubated with a suboptimal concentration of IL-5. This indicates that after allergen challenge, the bone marrow of the dual responders is more responsive to IL-5, which may reflect either a specific induction of a population of more committed eosinophil/basophil progenitors or an upregulation of the IL-5 receptor on the surface of these cells. Studies performed by Sehmi and colleagues (24) in our laboratory has confirmed an increase in the proportion of CD34+ cells expressing the alpha subunit of the IL-5 receptor after allergen in dual but not in isolated early responders, which supports the latter hypothesis. This increase in responsiveness to IL-5 in dual responders was associated with significant increases in airway eosinophils and methacholine airway responsiveness at 24 h after allergen challenge, which did not occur in the isolated early responders. These results may indicate that the responsiveness of the bone marrow to IL-5 after allergen is a determinant of the magnitude of the eosinophilic responses to inhaled allergen and of the degree of the subsequent physiologic abnormalities. Conversely, the degree of IL-5 sensitivity in the bone marrow may be determined by the events occurring in the airway that ultimately give rise to the LAR. However, these speculations will only be adequately addressed with the use of specific tools that block the activity of IL-5 on the bone marrow such as monoclonal antibodies directed against the cytokine.
Other studies have supported an important role for IL-5 in the development of allergen-induced airway inflammation. In studies of transgenic mice that overexpress IL-5, there is marked circulating and tissue eosinophilia (25), whereas in animal models, antibodies to IL-5 can block allergen-induced local and systemic eosinophilia and airway hyperresponsiveness for periods lasting as long as 6 mo. In murine, guinea-pig, and primate models, treatment of sensitized animals with the anti-IL-5 monoclonal antibody TRFK-5 inhibits the allergen-induced eosinophil infiltration into BAL fluid and lung tissue, and in primates the effect of this antibody could be detected for as long as 3 mo (26). In addition, TRFK-5 inhibited the increase in airway responsiveness in both the guinea-pig and primate models. A humanized anti-IL-5 antibody (Sch 55700) has been constructed, which has been shown to inhibit pulmonary eosinophilia in both sensitized guinea pigs and primates; the latter inhibitory effect on allergen-induced BAL eosinophilia also lasts as long as 6 mo (29), suggesting an ongoing effect on eosinophil production in the bone marrow.
The time point of 24 h used to measure eosinophil/basophil progenitors in the bone marrow was chosen based on the increased responsiveness of the bone marrow at 24 h after allergen inhalation in the canine model of allergen-induced AHR (14). Although increases were seen at this time point, this study does not provide any evidence on the timing of the bone marrow response. A more detailed kinetics study may provide further information on whether the bone marrow response after allergen is primary or secondary to events occurring in the airways.
The number of GM-CSF-stimulated GM-CFU colonies were also significantly higher in the dual responders after allergen inhalation. These results show that there is also a neutrophilic/monocytic progenitor response in the bone marrow after allergen inhalation; indeed, previous studies have implicated neutrophils during the late response, albeit transiently, just prior to development of eosinophilic inflammation (30, 31).
Previous studies from our laboratory, in a canine model of allergen-induced airway hyperresponsiveness and airway inflammation, have demonstrated that allergen inhalation increases bone marrow GM-CFU production (14), and that this is due to an as yet unidentified hemopoietic activity released into the bloodstream after allergen inhalation, which stimulates the bone marrow (32). The present study raises the possibility that after allergen inhalation by atopic asthmatics, IL-5 may represent one such hemopoietic signal with a significant effect on IL-5 responsive progenitors induced in the bone marrow; this could then lead to increased production of eosinophils and promote eosinophilic inflammation of the airways. Alternatively, a recent study by Minshall and colleagues (33) showed that after allergen inhalation, T-lymphocytes present in the bone marrow were a significant source of IL-5 in sensitized mice, suggesting that events occurring in the airways may result in local bone marrow production of IL-5, leading to increased production of eosinophils. Our studies in dogs have also shown that pretreatment with the inhaled corticosteroid, budesonide, can blunt the response of GM-CFU in the bone marrow after allergen inhalation (14). Thus, inhaled corticosteroids, which attenuate allergen-induced airway responses, as well as the increases in blood and airway eosinophils (34, 35), may partially exert their effect at the level of the bone marrow to prevent either eosinophil production or the release of maturing eosinophils into the bloodstream. These hypotheses require confirmation.
In conclusion, this study has demonstrated, for the first time, increases in human Eo/B and GM progenitors after allergen inhalation in sensitized, asthmatic subjects. However, an increased responsiveness of the Eo/B progenitors to suboptimal concentrations of IL-5 appears to distinguish between subjects who develop the most marked increases in airway eosinophils after allergen inhalation, which is associated with the development of allergen-induced late responses and airway hyperresponsiveness.
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Correspondence and requests for reprints should be addressed to Dr. Paul M. O'Byrne, Faculty of Health Sciences, Dept. of Medicine, 1200 Main St. W., Hamilton, ON, L8N 3Z5 Canada.
(Received in original form April 25, 1997 and in revised form July 29, 1997).
P. M. O'Byrne is a Medical Research Council of Canada Senior Scientist.Acknowledgments: The writers gratefully acknowledge the hematological expertise of Drs. Irwin Walker and Parveen Wassi.
Supported by the Medical Research Council of Canada.
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References |
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REFERENCES
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1. Cockcroft, D. W., and K. Y. Murdock. 1987. Changes in bronchial responsiveness to histamine at intervals after allergen challenge. Thorax 42: 302-308 [Abstract].
2. Gauvreau, G. M., J. Doctor, R. M. Watson, M. Jordana, and P. M. O'Byrne. 1996. Effects of inhaled budesonide on allergen-induced airway responses and airway inflammation. Am. J. Respir. Crit. Care Med. 154: 1267-1271 [Abstract].
3. Aalbers, R., H. F. Kauffman, G. H. Koeter, D. S. Postma, K. de Vries, and J. G. R. de Monchy. 1991. Dissimilarity in methacholine and AMP responsiveness 3 and 24 hours after allergen challenge. Am. Rev. Respir. Dis. 144: 352-357 [Medline].
4. Pin, I., A. P. Freitag, P. M. O'Byrne, A. Girgis-Girbardo, R. M. Watson, J. Dolovich, J. A. Denburg, and F. E. Hargreave. 1992. Changes in the cellular profile of induced-sputum after allergen-induced asthmatic responses. Am. Rev. Respir. Dis. 145: 1265-1269 [Medline].
5. Rossi, G. A., E. Crimi, S. Lanterno, P. Gianiorio, S. Oddera, P. Crimi, and V. Brusasco. 1991. Late phase asthmatic reaction to inhaled allergen is associated with early recruitment of eosinophils into the airways. Am. Rev. Respir. Dis. 144: 379-383 [Medline].
6. Bousquet, J., P. Chanez, J. Lacoste, G. Barreon, N. Ghavanian, I. Emander, P. Venge, S. Ahlstedt, J. Simony-Lafontaine, O. Goddard, and F. Michel. 1990. Eosinophilic inflammation in asthma. N. Engl. J. Med. 323: 1033-1039 [Abstract].
7. Denburg, J. A., S. Telizyn, A. Belda, J. Dolovich, and J. Bienenstock. 1985. Increased numbers of circulating basophil progenitors in atopic patients. J. Allergy Clin. Immunol. 76: 466-472 [Medline].
8. Denburg, J. A., J. Dolovich, and D. Harnish. 1989. Basophil mast cell and eosinophil growth and differentiation factors in human allergic disease. Clin. Exp. Allergy 19: 249-254 [Medline].
9. Sehmi, R., K. Howie, D. R. Sutherland, W. Schragge, P. M. O'Byrne, and J. A. Denburg. 1996. Increased levels of CD34+ hemopoietic progenitor cells in atopic subjects. Am. J. Respir. Cell Mol. Biol. 15: 645-654 [Abstract].
10. Gibson, P. G., J. Dolovich, A. Girgis-Girbado, M. Morris, M. Anderson, F. E. Hargreave, and J. A. Denburg. 1990. The inflammatory response in asthma exacerbation: changes in circulating eosinophils, basophils and their progenitors. Clin. Exp. Allergy 20: 661-668 [Medline].
11. Ohnishi, M., J. Ruhno, J. Dolovich, and J. A. Denburg. 1988. Allergic rhinitis nasal mucosal conditioned medium stimulates growth and differentiation of basophil/mast cell and eosinophil progenitors from atopic blood. J. Allergy Clin. Immunol. 81: 1149-1154 [Medline].
12. Otsuka, H., J. Dolovich, D. Befus, S. Telizyn, J. Bienenstock, and J. A. Denburg. 1986. Basophilic cell progenitors, nasal metachromatic cells, and peripheral blood basophils in ragweed-allergic patients. J. Allergy Clin. Immunol. 78: 365-371 [Medline].
13. Gibson, P. G., P. J. Manning, P. M. O'Byrne, A. Girgis-Girbado, J. Dolovich, J. A. Denburg, and F. E. Hargreave. 1991. Allergen-induced asthmatic responses: relationship between increases in airway responsiveness and increases in circulating eosinophils, basophils and their progenitors. Am. Rev. Respir. Dis. 143: 331-335 [Medline].
14. Woolley, M. J., J. A. Denburg, R. Ellis, M. Dahlback, and P. M. O'Byrne. 1994. Allergen-induced changes in bone marrow progenitors and airway responsiveness in dogs and the effect of inhaled budesonide on these parameters. Am. J. Respir. Cell Mol. Biol. 11: 600-606 [Abstract].
15. Cockcroft, D. W. 1985. Measure of airway responsiveness to inhaled histamine or methacholine: method of continuous aerosol generation and tidal breathing inhalation. In F. E. Hargreave and A. J. Woolcock, editors. Airway Responsiveness: Measurement and Interpretation. Astra Pharmaceuticals, Canada, Limited, Mississauga, ON. 22-28.
16. O'Byrne, P. M., J. Dolovich, and F. E. Hargreave. 1987. Late asthmatic response. Am. Rev. Respir. Dis. 136: 740-756 [Medline].
17. Cockcroft, D. W., K. Y. Murdock, J. Kirby, and F. E. Hargreave. 1987. Prediction of airway responsiveness to allergen from skin sensitivity to allergen and airway responsiveness to histamine. Am. Rev. Respir. Dis. 135: 264-267 [Medline].
18.
Denburg, J. A.,
S. Telizyn,
H. Messner,
B. L. N. Jamal,
S. J. Ackerman,
G. J. Gleich, and
J. Bienenstock.
1985.
Heterogeneity of human peripheral blood eosinophil type colonies: evidence for a common basophil-eosinophil progenitor.
Blood
66:
312-318
19. Popov, T. A., R. Gottschalk, R. Kilendowicz, J. Dolovich, J. Powers, and F. E. Hargreave. 1994. The evaluation of a cell dispersion method of sputum examination. Clin. Exp. Allergy 24: 778-783 [Medline].
20. Girgis-Girbado, A., N. Kanai, J. A. Denburg, F. E. Hargreave, M. Jordana, and J. Dolovich. 1994. Immunocytochemical detection of granulocyte-macrophage colony stimulating factor and eosinophil cationic protein in sputum cells. J. Allergy Clin. Immunol. 93: 945-947 [Medline].
21. Kleinbaum, D. G., L. L. Kupper, and K. E. Muller. 1981. Applied Regression Analysis and Other Multivariable Methods, 2nd ed. PWS-Kent, Boston.
22. Keppel, G. 1982. Design and Analysis: A Researcher's Handbook, 2nd ed. Prentice-Hall, Englewood Cliffs, NJ.
23. Takanashi, S., P. M. O'Byrne, J. Dolovich, and M. Jordana. 1995. Changes in eosinophils and IL-5 immunoreactivity in induced sputum and in PC20 in single and dual asthmatic responders (abstract). Am. J. Respir. Crit. Care Med. 151(4-6):A38.
24. Sehmi, R., L. J. Wood, R. M. Watson, P. M. O'Byrne, and J. A. Denburg. 1997. Increased expression of IL-5 receptor alpha-subunit on bone marrow derived CD34+ progenitors following allergen challenge in dual responder asthmatics (abstract). J. Allergy Clin. Immunol. 99: S272 .
25.
Sanderson, C. J.,
D. J. Warren, and
M. Strath.
1985.
Identification of a
lymphokine that stimulates eosinophil differentiation in vitro: its relationship to interleukin-3, and functional properties of eosinophils produced in culture.
J. Exp. Med.
162:
60-74
26. Mauser, P. J., A. M. Pitman, X. Fernandez, S. K. Foran, G. K. Adams III, W. Kreutner, R. W. Egan, and R. W. Chapman. 1995. Effects of an antibody to interleukin-5 in a monkey model of asthma. Am. J. Respir. Crit. Care Med. 152: 467-472 [Abstract].
27. Gulbenkian, A. R., R. W. Egan, X. Fernandez, H. Jones, W. Kreutner, T. Kung, F. Payvandi, L. Sullivan, J. A. Zurcher, and A. S. Watnick. 1992. Interleukin-5 modulates eosinophil accumulation in allergic guinea pig lung. Am. Rev. Respir. Dis. 146: 263-265 [Medline].
28. Mauser, P. J., A. Pitman, X. Fernandez, J. A. Zurcher, T. Kung, A. S. Watnick, R. W. Egan, W. Kreutner, and G. K. Adams III.. 1993. Inhibitory effect of the TRFK-5 anti-IL-5 antibody in a guinea pig model of asthma. Am. Rev. Respir. Dis. 148: 1623-1627 [Medline].
29. Egan, R. W., D. Athwahl, C. Chou, S. Emtage, C. Jehn, T. T. Kung, P. J. Mauser, N. J. Murgolo, and M. W. Bodmer. 1995. Inhibition of pulmonary eosinophilia and hyperreactivity by antibodies to interleukin-5. Int. Arch. Allergy Immunol. 107: 321-322 [Medline].
30. Weersink, E. J. M., D. S. Postma, R. Aalbers, and J. G. R. de Monchy. 1994. Early and late asthmatic reaction after allergen challenge. Respiration Medicine 88: 103-114 .
31. Diaz, P., C. Gonzalez, F. R. Galleguillos, and et al. . 1989. Leukocytes and mediators in bronchoalveolar lavage during allergen-induced late asthmatic reactions. Am. Rev. Respir. Dis. 139: 1383-1389 [Medline].
32. Inman, M. D., J. A. Denburg, R. Ellis, M. Dahlback, and P. M. O'Byrne. 1996. Allergen-induced increase in bone marrow progenitors in airway hyperresponsive dogs: regulation by a serum hemopoietic factor. Am. J. Respir. Cell Mol. Biol. 15: 305-311 [Abstract].
33. Minshall, E., R. Schleimer, L. Cameron, M. Minnicozzi, R. W. Egan, D. H. Eidelman, and Q. Hamid. 1997. IL-5 mRNA and immunoreactivity in the bone marrow of sensitized Balb/c mice following allergen challenge: evidence for T cell involvement (abstract). J. Allergy Clin. Immunol. 99: S165 .
34. Juniper, E. F., P. A. Kline, M. A. Vanzileghen, E. H. Ramsdale, P. M. O'Byrne, and F. E. Hargreave. 1990. Effect of long-term treatment with an inhaled corticosteroid (budesonide) on airway hyperresponsiveness and clinical asthma. Am. Rev. Respir. Dis. 142: 832-836 [Medline].
35. Laviolette, L., C. Ferland, L. Trepanier, H. Rocheleau, A. Dakhama, and L. Boulet. 1994. Effects of inhaled steroids on blood eosinophils in moderate asthma. Ann. N.Y. Acad. Sci. 725: 288-297 [Medline].
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