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ABSTRACT |
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INTRODUCTION
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Interleukin-5 (IL-5) is a potent eosinophilopoietic factor implicated in the chronic inflammatory cell accumulation accompanying bronchial asthma. However, its role in stimulating eosinophil differentiation within the bone marrow following allergen exposure remains to be elucidated. The aims of our study were to determine the expression of IL-5 within the bone marrow of sensitized and control mice after allergen exposure, and to investigate the cellular phenotype of IL-5-producing cells. Sensitized Balb/c mice were challenged with either ovalbumin (OVA) or sterile saline. After 6 h, the mice were exsanguinated and the bone marrow prepared for cytospins. Bone marrow-derived cells from OVA-sensitized mice exhibited an increase in IL-5 immunoreactivity and mRNA compared with those from nonsensitized control mice (p < 0.05). After allergen challenge, there was a further increase in IL-5 expression (p < 0.05) within the bone marrow. Both sensitization and allergen challenge resulted in an increase in the number of cells expressing major basic protein (MBP) (p < 0.05). In nonsensitized mice, the IL-5 mRNA was expressed predominantly by CD34-positive (CD34+) progenitor cells. Following sensitization and allergen challenge, CD3-positive (CD3+) T lymphocytes were the major source of this cytokine. These results demonstrate the presence of IL-5 within the bone marrow of normal Balb/c mice. After sensitization and allergen challenge, the increase in IL-5-producing cells within the bone marrow is attributed by T lymphocytes.
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INTRODUCTION |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
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Allergic inflammation is characterized by an eosinophilic infiltration of the tissues (1, 2). The presence of these cells and the subsequent liberation of their inflammatory mediators has been linked to the pathophysiology of atopic disorders such as allergic asthma (3). Acute administration of allergen into the airways of asthmatics elicits eosinophil infiltration which is associated with the local release of chemokines (6), as well as the activation of adhesion molecules (7). To support the recruitment of eosinophils into the tissues, there is an early release of eosinophils into the circulation (8), and subsequently an enhanced production of these cells within the bone marrow (8, 9). While the mechanisms responsible for the trafficking of eosinophils to the airways and their subsequent activation in situ have been extensively examined, there has been very little progress to date understanding the mechanisms regulating the production of eosinophils in vivo.
Eosinophilopoiesis occurs predominantly in the bone marrow, where pluripotent stem cells terminally differentiate along the eosinophilic lineage in the presence of interleukin-5 (IL-5) (10, 11). Evidence to support the role of IL-5 in vivo has been obtained from transgenic mice, where overexpression of IL-5 leads to a massive peripheral blood and bone marrow eosinophilia (9, 12). Moreover, neutralization of its bioactivity using specific anti-IL-5 antibodies inhibits the production of these cells (13, 15). Although eosinophils are found constitutively within the bone marrow, the localization of eosinophilopoietic factors such as IL-5 has not been studied to date. Furthermore, while eosinophil numbers have been shown to be acutely elevated within the bone marrow in response to allergen exposure (8, 9), the concomitant increased production of IL-5 remains to be investigated.
We hypothesized that there is a constitutive expression of IL-5 within the bone marrow responsible for the production of eosinophils, and that allergic sensitization and subsequent exposure of the airways to allergen would enhance this expression. Therefore, the aim of our study was to determine the numbers of cells within the bone marrow expressing IL-5 messenger RNA (mRNA) and immunoreactivity in response to sensitization and allergen-challenge, using the techniques of in situ hybridization (ISH) and immunocytochemistry (ICC). To determine whether this was associated with an increased production of eosinophils, we also investigated the percentage of major basic protein-positive (MBP+) cells within the bone marrow. In addition, we phenotyped the cells expressing IL-5 mRNA in normal bone marrow, and after allergic sensitization and challenge.
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ABSTRACT
INTRODUCTION
METHODS
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DISCUSSION
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Animals
Male Balb/c mice (6- to 8-wk-old) were obtained from a commercial supplier (Jackson Laboratories, Bar Harbor, ME) and housed in a conventional animal facility at our laboratory. Twelve animals were sensitized and six served as normal, nonsensitized control mice. Animals were studied 14 d after allergen-sensitization. All methods used in these experiments were evaluated and approved by the animal ethics committee at McGill University.
Antigen Sensitization and Challenge
Balb/c mice were sensitized to ovalbumin (OVA) according to a modification of a protocol previously demonstrated to elicit airways inflammation and late airways responses in rats (16). Briefly, we administered 100 µg OVA (Sigma, St. Louis, MO) and 0.43 mg aluminum hydroxide gel (BHD Chemicals, Poole, UK) in 0.5 ml of saline subcutaneously, and 0.5 ml Bordetella pertussis vaccine (IAF BioVac, Laval, PQ, Canada) with 2 × 10 9 heat-killed organisms intraperitoneally as an adjuvant. Fourteen days after sensitization, intratracheal challenge with either OVA or sterile saline was carried out in OVA-sensitized mice. The mice were sedated with xylazine (8 mg/kg, intraperitoneally), anesthetized with sodium pentobarbital (70 mg/kg, intraperitoneally), and then restrained on a board at a fixed angle. Mice were then randomly assigned to receive either OVA (100 µg; 50 µl) or sterile saline (50 µl) by intratracheal instillation using a blunt-ended 21G needle. The mice were then allowed to recover from the anesthesia.
Bone Marrow Cytospin Preparation
Bone marrow samples were taken from the right femur for ISH and
ICC 6 h after allergen or saline challenge. The mice were anesthetized
(sodium pentobarbital; 60 mg/kg), exsanguinated, and the bone marrow was flushed using 2.5 ml sterile saline containing 100 U/ml heparin. Cytospin preparations (approximately 10 5 cells) of the cell suspension were prepared on either poly-L-lysine or uncoated slides for
ISH and ICC, respectively. For in situ hybridization, the cytospins
were fixed in 4% paraformaldehyde for 30 min and then twice washed
in phosphate-buffered saline (PBS). The slides were stored at 
80° C
prior to use. For immunocytochemistry, the slides were briefly air
dried, fixed in acetone:methanol for 7 min, and stored at 20° C before use.
In Situ Hybridization
To detect mouse IL-5 mRNA, the technique of ISH using a digoxigenin-labeled complementary RNA (cRNA) probe was used as previously described (17). The probe was generated from complementary DNA (cDNA) (a kind gift of Dr. T. Honjo, Kyoto University, Japan) and transcribed in the presence of SP6 or T7 polymerases, and labeled with digoxigenin-11-uridine triphosphate (UTP) to generate antisense and sense probes, respectively. The bone marrow cytospins were permeabilized with proteinase K and then prehybridized with 50% formamide and 2× standard saline citrate. Following application of the probe, the sections were hybridized at 42° C overnight. Nonspecific binding was removed by posthybridization washing under high stringency conditions and subsequent treatment with ribonuclease (RNase). The hybridization signal was visualized by incubating the cytospins overnight with sheep polyclonal antidigoxigenin antibodies conjugated with alkaline phosphatase. Color development was achieved by adding the freshly prepared substrate (X-phosphate-5-bromo-4-chloro-3-indoly phosphate and nitroblue tetrazolium). To ensure the specificity of our signal, we performed the ISH technique using the sense probe, and following pretreatment of the tissues with RNase.
Immunocytochemistry
IL-5 and MBP immunoreactivity were evaluated in bone marrow cell cytospins of allergen-immunized and normal mice using monoclonal and polyclonal antibodies specific for mouse IL-5 (TRFK-5; Schering-Plough Research Institute, New Jersey) and mouse MBP (a kind gift of Dr. G. Gleich, Mayo Clinic, Rochester, MN), respectively. For the negative control preparations, the primary antibody was replaced with either a rat anti-mouse isotype control antibody from GL-113 cells, a rabbit anti-mouse isotype control antibody, or TRIS-buffered saline (TBS). To detect differences in the cellular phenotype, we also evaluated the numbers of T lymphocytes and progenitor cells in these bone marrow samples. These were identified using polyclonal antibodies against specific cell surface markers for mouse T lymphocytes (hamster anti-murine CD3; Dako Diagnostics Canada, Inc., Mississauga, ON, Canada) and progenitor cells (rat antimurine CD34, 18). For the control antibody staining, the primary antibody was replaced with the appropriate hamster or rat anti-mouse isotype or TBS. Numbers of IL-5 and MBP-immunoreactive cells were identified by the technique of alkaline phosphatase-anti-alkaline phosphatase (APAAP) immunocytochemistry and those expressing CD3 and CD34 by the avidin-biotin complex (ABC) method, as previously described (19, 20). Using this procedure, cells expressing IL-5 or MBP immunoreactivity were stained red and those exhibiting cell surface markers for CD3 or CD34 stained brown.
Combined In Situ Hybridization and Immunocytochemistry
To ascertain the expression of IL-5 mRNA by T lymphocytes and progenitor cells, we simultaneously applied radiolabeled in situ hybridization with immunocytochemistry for CD3 and CD34 as previously described in detail (21). Briefly, cytospin preparations were immunostained with monoclonal antibodies directed against cell surface markers specific for T lymphocytes and progenitor cells using the ABC technique. After visualization of the positive immunostaining, the sections underwent a modified ISH for IL-5 mRNA using a specific radiolabeled cRNA probe for murine IL-5.
Histological Analysis
Slides were randomly encoded and read by two observers blinded to the identity of the slides. The interobserver variability was less than 10%. IL-5 mRNA and immunoreactive-positive cells, MBP-immunoreactive positive cells as well as those positive for CD3 and CD34 were counted as a percentage of the total number of bone marrow cells. The numbers of CD34+ IL-5 mRNA-positive (mRNA+) cells were expressed both as the percentage total CD34+ and the percentage total IL-5 mRNA+ cells. A similar analysis was applied to the numbers of cells expressing both CD3 and IL-5 mRNA. At least 1,000 total cells were counted per slide and 2 slides were hybridized for each animal. The results are expressed as the mean percentage of mRNA or immunoreactive-positive cells/total cells.
Statistical Analysis
For cell counts within the bone marrow, the data are expressed as the mean percentage of the total cell count ± SEM. In the colocalization studies involving the phenotyping of IL-5 mRNA+ cells, the data are given as a percentage of the total CD3 and CD34+ cells. The data were analyzed using a nonparametric Kruskal-Wallis analysis of variance with subsequent post hoc Mann-Whitney U tests (SyStat v6.1; SPSS Inc., Chicago, IL). The coefficients of determination (r2) and significance values were calculated using Pearson's product-moment correlation and Bartlett's test (Systat v5.0). p Values less than 0.05 were considered statistically significant.
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INTRODUCTION
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DISCUSSION
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Using APAAP immunostaining, IL-5-immunoreactive positive cells exhibited red cytoplasmic staining overlying cells within the bone marrow of nonsensitized and sensitized mice (Figures 1a and 1b). Cells positive for IL-5 mRNA exhibited a dark purple staining within the bone marrow of nonsensitized (Figure 1c) and OVA-sensitized mice (Figures 1d and 1e) by digoxigenin-labeled in situ hybridization. IL-5-immunoreactivity was not observed when the control isotype primary antibody was used (Figure 1f). Moreover, the detection of IL-5 mRNA was observed only with the antisense probe, confirming the specificity of our hybridization technique. The colocalized expression of IL-5 mRNA with CD34+ and CD3+ cells following allergen challenge in sensitized mice can be seen in the insets in Figures 1c and 1d. Cells expressing IL-5 mRNA that were CD34+ and CD3+ could be identified by the presence of silver grains within cells exhibiting brown immunostaining for the appropriate cell surface markers.
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IL-5 mRNA Expression Following Sensitization and Allergen Challenge
Our results show that there is constitutive expression of IL-5 mRNA within the bone marrow of normal, nonsensitized animals (Figure 2, 1.7 ± 0.35%, n = 6). The bone marrow from OVA-sensitized animals receiving sterile saline exhibited a significant increase in the percentage of cells expressing IL-5 mRNA (5.55 ± 0.83, n = 6; p < 0.05). Six hours after allergen challenge, there was a further significant increase in the percentage of IL-5 mRNA+ cells compared with the saline-challenged sensitized animals (14.1 ± 1.9, n = 6; p < 0.05).
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IL-5 Immunoreactivity Following Sensitization and Allergen Challenge
To confirm that the IL-5 protein was also expressed within the bone marrow, we used APAAP immunocytochemistry to detect IL-5 immunoreactivity. The profile of IL-5 immunoreactivity was similarly increased in the sensitized mice receiving sterile saline compared with the normal control mice (Figure 3; nonsensitized, 2.47 ± 0.55, n = 6; sensitized saline-challenged, 5.02 ± 0.46, n = 6; p < 0.05). There was also a further increase in IL-5 immunoreactivity in the sensitized allergen-challenged group of mice compared with those challenged with sterile saline (8.91 ± 1.47, n = 6; p < 0.05). When we performed a correlational analysis in the three groups, there was a strong association between the numbers of IL-5 mRNA and IL-5-immunoreactive cells (r2 = 0.87, p < 0.05).
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MBP Immunoreactivity Following Sensitization and Allergen Challenge
To detect the percentage of eosinophils within the bone marrow of nonsensitized mice and after sensitization and allergen challenge, we performed APAAP immunostaining for MBP. In normal mice, there was a constitutive expression of MBP within the bone marrow (Figure 4, 1.7 ± 0.35; n = 6). After sensitization and saline challenge, there was a significant increase in the percentage of cells expressing MBP (5.56 ± 0.83, n = 6; p < 0.05). This percentage was further increased almost threefold in the mice that received allergen challenge (14.1 ± 1.9; n = 6; p < 0.05). When a correlational analysis was performed in the three groups, there were significant associations between the numbers of cells expressing MBP and the expression of IL-5 mRNA (r2 = 0.69) and IL-5-immunoreactivity (r2 = 0.62).
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Phenotype of IL-5 mRNA Expressing Cells in Response to Sensitization and Allergen Challenge
To ascertain the phenotype of cells within the bone marrow expressing IL-5 mRNA, we performed colocalization studies with immunocytochemical markers of T lymphocytes (CD3) and progenitor cells (CD34). These results (Table 1) indicate that in normal nonsensitized mice the majority of IL-5 mRNA is associated with CD34+ progenitor cells; however, there are small numbers of IL-5 mRNA+CD3+ T cells within the bone marrow. Following sensitization, there was a significant increase in the percentage of CD3+ T cells expressing IL-5 mRNA (Table 1; p < 0.03). In contrast to the observed increase in the percentage of T lymphocytes expressing IL-5 mRNA, sensitized saline-challenged mice displayed a significant decrease in the percentage of CD34+ cells expressing this cytokine mRNA (Table 1; p < 0.03).
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After allergen challenge in OVA-sensitized mice, there was a significant increase in the percentage of CD3+ T lymphocytes expressing IL-5 mRNA when compared with saline-challenged sensitized animals (Table 1; p < 0.03). Similarly, compared with OVA-sensitized saline-challenged mice, allergen-challenged mice exhibited a significant increase in the percentage of stem cells expressing IL-5 mRNA (Table 1; p < 0.05). These values were similar to those observed in nonsensitized animals.
Numbers of CD3+ and CD34+ Cells in the Bone Marrow and Their IL-5 mRNA Expression
The numbers of CD3 T lymphocytes and CD34 progenitor cells were examined within the bone marrow of normal and sensitized mice to determine whether the changes in IL-5 expression could be attributed to alterations in the numbers of inflammatory cells present (Table 2). In saline-challenged mice, there was no increase in the number of CD3+ cells when compared with nonsensitized control mice. In contrast, allergen challenge in OVA-sensitized mice was associated with a greater than fourfold increase in the percentage of T lymphocytes within the bone marrow compared with saline-challenged animals (p < 0.01). In saline-challenged sensitized mice, there was a threefold increase in the numbers of CD34+ cells compared with the nonsensitized control animals (Table 2; p < 0.01). Compared with these saline-challenged mice, an approximate doubling in the numbers of CD34+ cells was observed in animals exposed to OVA (p < 0.05).
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When the numbers of CD3+ and CD34+ cells expressing IL-5 mRNA were determined, it could be seen that over 50% of CD3+ T lymphocytes within the bone marrow of allergen-challenged mice expressed this cytokine mRNA (Table 2). This was significantly greater than that observed in the saline-challenged group (p < 0.05). Similarly, over 20% of all CD34+ cells in the bone marrow of allergen-challenged mice expressed IL-5 mRNA and this was approximately threefold greater than that seen in the saline-challenged group (p < 0.05).
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DISCUSSION |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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We undertook to determine the expression of IL-5 mRNA and immunoreactivity within the bone marrow of Balb/c mice after sensitization and allergen challenge and to characterize the phenotype of cells expressing cytokine mRNA. To assess whether there was a physiological role for IL-5 production in the bone marrow, we also examined the relationship between IL-5 expression and the numbers of MBP+ eosinophils following allergen stimulation. Our results demonstrated that there is a constitutive expression of IL-5 within the bone marrow of normal mice. This expression is increased after sensitization and allergen challenge and correlated to the numbers of MBP+ eosinophils. These data also indicate that progenitor cells expressing CD34 are the major source of IL-5 mRNA during unstimulated eosinophilopoiesis, and that CD3+ T lymphocytes make a major contribution to the expression of this cytokine particularly after sensitization and allergen challenge.
Studies performed in vitro have shown that eosinophilopoiesis is under the influence of several cytokines within the bone marrow including IL-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF), however it appears to be critically dependent upon the presence of IL-5 within the local milieu (10, 11). As such, the production of eosinophils is severely compromised in IL-5 knockout mice (22, 23), and hypereosinophilia is a characterisic feature of both IL-5 transgenic animals (9, 12) and IL-5 given systemically (24). Our results in normal, nonsensitized mice show that IL-5 is constitutively expressed in the bone marrow, predominantly by CD34+ progenitor cells. Because these cells are precursors for all blood leukocytes, including eosinophils, there is the possibility that IL-5 exhibits an autocrine action. Thus, the expression of this cytokine at rest is possibly an initiating factor for commitment to the eosinophil/basophil lineage. Further studies examining the distribution of the IL-5 receptor on stem cells during eosinophilopoiesis are necessary to confirm this hypothesis.
Sensitization of Balb/c mice with OVA resulted in an increased IL-5 mRNA and immunoreactive protein expression within the bone marrow. This increase in IL-5 expression within the bone marrow may augment the terminal differentiation of eosinophils and thus prime the immune system for subsequent exposure to allergen. Indeed, our results confirmed previous observations in demonstrating that the numbers of eosinophils were increased within the bone marrow and blood as a result of OVA sensitization (9, 25). Interestingly, the increase in IL-5 mRNA expression we observed in the bone marrow of saline-challenged sensitized mice was associated with CD3+ T cells. In the absence of an increase in the percentage of T cells found within the marrow (Table 2), these data suggest OVA sensitization induces switching of these cells in situ to favor IL-5 production. However, we can not dismiss the possibility that CD3+ cells expressing IL-5 mRNA migrate into the bone marrow after sensitization, and take the place of T lymphocytes not expressing IL-5 mRNA.
To determine the numbers of progenitor cells within the bone marrow and their expression of IL-5 mRNA, we used an antibody directed against the CD34 cell surface marker. It is known that this antibody will recognize a variety of cell types in addition to stem cells, notably fibroblasts and endothelial cells (18). While we have attempted to use alternative means of discriminating these cells types in the mouse, the specific immunological reagents are, as yet, not commercially available. As such, it is impossible to know exactly what contribution these individual cell types are having to the production of IL-5 in the mouse. Nevertheless, there is evidence in the literature to suggest that the majority of CD34+ cells within the bone marrow of the mouse are progenitor cells (26). Therefore, it may be expected that these CD34+ cells expressing IL-5 mRNA are indeed hemopoietic progenitors.
Compared with naive mice, OVA-sensitized animals showed a significant decrease in the numbers of CD34+ cells expressing IL-5 mRNA. One explanation of this finding is that these progenitor cells had left the bone marrow and entered the peripheral circulation. Indeed, there was a significant increase in the percentage of CD34+ cells after sensitization. As such, our observations are consistent with the increased numbers of CD34+ cells found within the blood of atopic individuals (27). However, the decrease in IL-5 mRNA+CD34+ cells may also be a consequence of IL-5-induced terminal differentiation, leading to an inability of these cells to express IL-5 mRNA. Further studies examining the presence of CD34+ progenitor cells within the blood following sensitization are necessary to address this issue.
Six hours after allergen challenge, the numbers of IL-5 mRNA and immunoreactive cells in the bone marrow were significantly increased. The percentages of cells expressing IL-5 mRNA and immunoreactivity were closely correlated and the presence of such an association suggests transcriptional regulation of this cytokine within the bone marrow, as has previously been described for IL-5 (28, 29). While we demonstrated that allergen challenge resulted in an increase in the numbers of MBP+ eosinophils within the bone marrow, eosinophilia within the bone marrow, blood, and lung tissue has previously been shown to accompany allergen challenge in sensitized mice (8, 9, 13, 14). Using our data, we found close correlations between the percentage of MBP+ eosinophils and the percentages of IL-5 mRNA and immunoreactive cells. These studies would suggest that not only is IL-5 being transcribed and translated in the bone marrow as a result of allergic stimulation, but it also exerts a biological action in inducing the differentiation of eosinophil precursors.
The immunological changes occurring within the bone marrow were in response to allergen challenge of the airways. These observations suggest that there is a mechanism of communication between antigen-induced inflammatory events within the tissue and the bone marrow. Indeed, there is preliminary evidence to support the presence of a factor released by the lung on allergen challenge which stimulates the bone marrow to increase the production of granulocytes (30). The nature of this factor and its relationship to allergic disorders in humans remains to be established. When we examined the phenotype of cells expressing IL-5 after allergen challenge, it was observed that T lymphocytes comprised approximately 50% of the IL-5 mRNA+ cells. This was associated with a fourfold increase in the percentages of T lymphocytes found within the bone marrow. As such, our data would indicate that local allergen challenge to the airways elicits the recruitment of T lymphocytes to the bone marrow. Indeed, mobilization of T lymphocytes to the tissues and lymph nodes following allergen challenge in sensitized animals and during parasitic infections has been previously described (14, 31). Although the phenotype of these T cells remains to be determined, a variety of studies have demonstrated the importance of CD4+ (helper) T lymphocytes in allergen-induced eosinophil recruitment (14, 33, 34). Despite the evidence for T-cell infiltration, it is impossible to rule out the possibility that a local proliferation of lymphocytes occurred within the bone marrow following allergen challenge, and that it is these cells which are expressing IL-5 mRNA.
In summary, we have shown that there is a constitutive expression of IL-5 within the bone marrow which is regulated by both IL-5-producing progenitors and T cells. Furthermore, we have determined that the presence of this cytokine is augmented by both OVA sensitization and allergen challenge. This increase in IL-5 mRNA expression can be attributed to the switching and recruitment of T cells, respectively. These results suggest a role for bone marrow-derived IL-5 in the pathophysiology of eosinophil-related disorders and help clarify the complex interaction between T cells and eosinophils.
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Footnotes |
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Correspondence and requests for reprints should be addressed to Dr. Qutayba Hamid, M.D., Ph.D., Meakins-Christie Laboratories, McGill University, 3626 St. Urbain, Montreal, PQ, H2X 2P2 Canada.
(Received in original form September 25, 1997 and in revised form April 10, 1998).
Dr. Minshall is a recipient of a Medical Research Council/Canadian Lung Association Fellowship.Drs. Eidelman and Hamid are recipients of Chercheur-Boursier awards from the Fonds de la Recherche en Santé du Quebec.
Acknowledgments: The authors thank Elsa Schotman, Zivart Yasruel, and Kevin Kembal for their invaluable technical assistance.
Supported by MRC Canada, Inspiraplex, and the J. T. Costello Memorial Research Foundation.
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References |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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1. Bradley, B. L., M. Azzawi, M. Jacobson, B. Assoufi, J. V. Collins, A.-M. A. Irani, L. B. Schwartz, S. R. Durham, P. K. Jeffery, and A. B. Kay. 1991. Eosinophils, T-lymphocytes, mast cells, neutrophils, and macrophages in bronchial biopsy specimens from atopic subjects without asthma and normal control subjects and relationship to bronchial hyperresponsiveness. J. Allergy Clin. Immunol 88: 661-674 [Medline].
2. Azzawi, M., B. Bradley, P. K. Jeffery, A. J. Frew, A. J. Wardlaw, B. Assoufi, J. V. Collins, S. R. Durham, G. K. Knowles, and A. B. Kay. 1990. Identification of activated T lymphocytes and eosinophils in bronchial biopsies in stable atopic asthma. Am. Rev. Respir. Dis 142: 1407-1413 [Medline].
3. Frigas, E., S. Motojima, and G. J. Gleich. 1991. The eosinophilic injury to the mucosa of the airways in the pathogenesis of bronchial asthma. Eur. Respir. J 4: S123-S135 .
4. Gundel, R. H., G. Letts, and G. J. Gleich. 1991. Human eosinophil major basic protein induces airway constriction and airway hyperresponsiveness in primates. J. Clin. Invest. 1470-1473.
5. Bascom, R., U. Pipkorn, D. Proud, S. Dunnette, G. J. Gleich, L. M. Lichtenstein, and R. M. Naclerio. 1989. Major basic protein and eosinophil-derived neurotoxin concentrations in nasal-lavage fluid after allergen challenge: effect of systemic corticosteroids and relationship to eosinophil influx. J. Allergy Clin. Immunol 84: 338-346 [Medline].
6. Lamkhioued, B., P. M. Renzi, S. Abi-Younes, E. A. Garcia-Zepada, A. D. Luster, and Q. A. Hamid. 1997. Increased expression of eotaxin in BAL and airways of asthmatics contributes to the chemotaxis of eosinophils to the site of inflammation. J. Immunol 159: 4593-4601 [Abstract].
7. Fukuda, T., Y. Fukushima, T. Numao, N. Ando, M. Arima, H. Nakajima, H. Sagara, T. Adachi, S. Motojima, and S. Makino. 1996. Role of interleukin-4 and vascular cell adhesion molecule-1 in selective eosinophil migration into the airways in allergic asthma. Am. J. Respir. Cell Mol. Biol 14: 84-94 [Abstract].
8. Kung, T. T., D. Stelts, J. A. Zurcher, A. S. Watnick, H. Jones, P. J. Mauser, X. Fernandez, S. Umland, W. Kreutner, R. W. Chapman, and R. W. Egan. 1994. Mechanisms of allergic pulmonary eosinophilia in the mouse. J. Allergy Clin. Immunol 94: 1217-1224 [Medline].
9. Lefort, J., C.-M. Bachelet, D. Leduc, and B. B. Vargaftig. 1996. Effect of antigen provocation of IL-5 transgenic mice on eosinophil mobilization and bronchial hyperresponsiveness. J. Allergy Clin. Immunol 97: 788-799 [Medline].
10.
Clutterbuck, E. J.,
E. M. A. Hirst, and
C. J. Sanderson.
1989.
Human interleukin-5 (IL-5) regulates the production of eosinophils in human
bone marrow cultures: comparison and interaction with IL-1, IL-3, IL-6
and GM-CSF.
Blood
73:
1504-1512
11.
Yamaguchi, Y.,
T. Suda,
J. Suda,
M. Eguchi,
Y. Miura,
N. Harad,
A. Taminaga, and
K. Takatsu.
1988.
Purified interleukin 5 supports the
terminal differentiation and proliferation of murine eosinophilic precursors.
J. Exp. Med
167:
43-56
12.
Dent, L. A.,
M. Strath,
A. L. Mellor, and
C. J. Sanderson.
1990.
Eosinophilia in transgenic mice expressing interleukin 5.
J. Exp. Med
172:
1425-1431
13. Kung, T. T., D. M. Stelts, J. A. Zurcher, G. K. Adams III, R. W. Egan, W. Kreutner, A. S. Watnick, H. Jones, and R. W. Chapman. 1995. Involvement of IL-5 in a murine model of allergic pulmonary inflammation: prophylactic and therapeutic effect of an anti-IL-5 antibody. Am. J. Respir. Cell Mol. Biol 13: 360-365 [Abstract].
14. Nakajima, H., I. Iwamoto, S. Tomoe, R. Matsumura, H. Tomioka, K. Takatsu, and S. Yoshida. 1992. CD4+ T-lymphocytes and interleukin-5 mediate antigen-induced eosinophil infiltration into the mouse trachea. Am. Rev. Respir. Dis 146: 374-377 [Medline].
15. Buijs, J., M. W. E. C. Egbers, W. H. Lokhorst, H. F. J. Savelkoul, and F. P. Nijkamp. 1995. Toxocara-induced eosinophilic inflammation: airway function and effect of anti-IL-5. Am. J. Respir. Crit. Care Med 151: 873-878 [Abstract].
16. Dandurand, R. J., C. G. Wang, S. Laberge, J. G. Martin, and D. H. Eidelman. 1994. In vitro allergic bronchoconstriction in the Brown Norway rat. Am. J. Respir. Crit. Care Med 149: 1499-1505 [Abstract].
17. Hamid, Q., M. Azzawi, S. Ying, R. Moqbel, A. J. Wardlaw, C. J. Corrigan, B. Bradley, S. R. Durham, J. V. Collins, P. K. Jeffery, D. J. Quint, and A. B. Kay. 1991. Expression of mRNA for interleukin-5 in mucosal bronchial biopsies from asthma. J. Clin. Invest 87: 1541-1546 .
18. Lin, G., E. Finger, and J. C. Gutierrez-Ramos. 1995. Expression of CD34 in endothelial cells, hematopoietic progenitors and nervous cells in fetal and adult mouse tissues. Eur. J. Immunol 25: 1508-1516 [Medline].
19. Durham, S. R., S. Ying, V. A. Varney, M. R. Jacobson, R. M. Sudderick, I. S. Mackay, A. B. Kay, and Q. Hamid. 1992. Cytokine messenger mRNA expression for IL-3, IL-4, IL-5 and granulocyte/macrophage colony-stimulating factor in the nasal mucosa after local allergen provocation: relationship to tissue eosinophilia. J. Immunol 148: 2390-2394 [Abstract].
20. Giaid, A., R. P. Michel, D. J. Stewart, M. Sheppard, B. Corrin, and Q. Hamid. 1993. Expression of endothelin-1 in lungs of patients with cryptogenic fibrosing alveolitis. Lancet 341: 1550-15554 [Medline].
21.
Hamid, Q.,
J. Barkans,
Q. Meng,
J. Abrams,
A. B. Kay, and
R. Moqbel.
1992.
Human eosinophils synthesize and secret interleukin-6, in vitro.
Blood
80:
1496-1501
22. Kopf, M., F. Brombacher, P. D. Hodgkin, A. J. Ramsay, E. A. Milbourne, W. J. Dai, K. S. Ovington, C. A. Behm, G. Kohler, I. G. Young, and K. I. Matthaei. 1996. IL-5 deficient mice have a developmental defect in CD5+ B-1 cells and lack eosinophilia but have normal antibody and cytotoxic T cell responses. Immunity 4: 15-24 [Medline].
23.
Foster, P. S.,
S. P. Hogen,
A. J. Ramsay,
K. I. Matthaei, and
I. G. Young.
1996.
Interleukin 5 deficiency abolishes eosinophilia, airways hyperreactivity and lung damage in a mouse asthma model.
J. Exp. Med
183:
195-201
24. Mould, A. W., K. I. Matthaei, I. G. Young, and P. S. Foster. 1997. Relationship between interleukin-5 and eotaxin in regulating blood and tissue eosinophilia in mice. J. Clin. Invest 99: 1064-1071 [Medline].
25. Takenaka, T., K. Kuribayashi, H. Nakamine, M. Tsujimoto, Y. Fukuhara, J. Maeda, M. Mihara, Y. Uchiyama, and Y. Ohsugi. 1993. Regulation by cytokines of eosinophilopoiesis and immunoglobulin E production in mice. Immunology 78: 541-546 [Medline].
26. Ito, A., S. Nomura, S. Hirota, J. Suda, T. Suda, and Y. Kitamura. 1995. Enhanced expression of CD34 messenger RNA by developing endothelial cells of mice. Lab. Invest 72: 532-538 [Medline].
27. Sehmi, R., K. Howie, D. R. Sutherland, W. Schragge, P. M. O'Byrne, and J. Denburg. 1996. Increased levels of CD34+ hemopoietic progenitor cells in atopic subjects. Am. J. Respir. Cell Mol. Biol. 15: 645-654 [Abstract].
28.
Foissier, L.,
M. Lonchampt,
F. Coge, and
E. Canet.
1996.
In vitro down-regulation of antigen-induced IL-5 gene expression and protein production by cAMP-specific phosphodiesterase type 4 inhibitor.
J. Pharmacol. Exp. Ther
278:
1484-1490
29.
Naora, H., and
I. G. Young.
1994.
Mechanisms regulating the mRNA
levels of interleukin-5 and two other co-ordinately expressed lymphokines in the murine T lymphoma EL4.23.
Blood
83:
3620-3628
30. 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].
31. Gonzalo, J.-A., C. M. Lloyd, L. Kremer, E. Finger, C. Martinez-A., M. H. Siegelman, M. Cybulsky, and J.-C. Gutierrez-Ramos. 1996. Eosinophil recruitment to the lung in a murine model of allergic inflammation. J. Clin. Invest 98: 2332-2345 [Medline].
32. Garlisi, C. G., A. Falcone, T. T. Kung, D. Stelts, K. J. Pennline, A. J. Beavis, S. R. Smith, R. W. Egan, and S. P. Umland. 1995. T cells are necessary for Th2 cytokine production and eosinophil accumulation in airways of antigen-challenged allergic mice. Clin Immunol. Immunopathol 75: 75-83 [Medline].
33. Watkins, A. D., C. A. Hatfield, S. F. Fidler, G. E. Winterrowd, J. R. Brashler, F. F. Sun, B. M. Taylor, S. L. Vonderfecht, G. A. Conder, S. T. Holgate, J. E. Chin, and I. M. Richards. 1996. Phenotypic analysis of airway eosinophils and lymphocytes in a Th-2-driven murine model of pulmonary inflammation. Am. J. Respir. Cell Mol. Biol 15: 20-34 [Abstract].
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