Waseem Saeed*, Ahmed Badar, M Mazhar Hussain, Muhammad Aslam
*Department of Pulmonology, Military Hospital and Department of Physiology, Army Medical College, Rawalpindi, Pakistan
Eosinophils are known to be an indirect marker of airway inflammation in asthma. It is known since long that the total eosinophil count reflects asthmatic activity and is useful for regulating steroid dosage and for early detection of exacerbations. Eosinophils are currently regarded as the effector cells responsible for much of the pathology of asthma. Eosinophil-mediated damage to the respiratory epithelium is a major pathogenetic mechanism in asthma. This article is a review of the latest works about the relationship of eosinophil and eosinophil products with asthma.
Asthma is an inflammatory disease of the airways characterized by its reversible, reactive nature. Asthma is not progressive, but is rather notable for episodes of exacerbations and remissions. Its symptoms are a result of airflow obstruction and include dyspnea, cough, wheezing, and chest tightness. The classic triad consists of cough, wheeze, and dyspnea. Traditionally, symptoms are worse at night or early in the morning 1. A number of hypotheses about the pathogenesis of asthma have been suggested that propose totally different mechanisms of causality at the biological level. These can be grouped into allergic, inflammatory, neurogenic, and physical mechanisms with current evidence in favour of a combination of allergic and inflammatory processes.
The obstruction of airways in acute fatal asthma is due to contraction of bronchial smooth muscle, thickening of airway wall and intraluminal mucus and debris. The wall thickening is characterised by submucosal oedema, vasodilatation, with cellular infiltrates, predominantly of lymphocytes and eosinophils. The lumen contains mucus plugs, shed epithelium (Creola bodies) and Charcot-Leyden crystals (crystallised eosinophil derived major basic protein). Bronchoscopic biopsies show presence of significantly more active eosinophils in the submucosae of mild and even asymptomatic asthmatics compared to controls. T lymphocytes are also present in increased numbers. Airway inflammation is present during an acute exacerbation of asthma, and is characterized by infiltration and activation of both eosinophils and neutrophils2.
Lavage fluid from the lumen of the asthmatic airway contains numerous mediators including histamine, prostaglandins (predominantly PGD2 and PGF2), leukotrienes, kinins, kallikrein, and eosinophil derived major basic protein. In the current model of asthma as an allergic disease, two cell types are central. These are the lymphocyte, especially in the role of T-cell as a regulator of immune processes, and the eosinophil, as effector cell causing tissue damage3,4.
Eosinophils are involved in defence against parasites but are best known for their role in allergic diseases and asthma5. A prominent feature of the eosinophil is the presence of many spherical or ovoid granules in their cytoplasm. Eosinophilic granulocytes are so named because they are stained intensely by eosin. Under the light microscope, a bilobed nucleus is typically seen in a normal healthy eosinophil6. Eosinophils are uniquely endowed with an arsenal of enzymes that enables them to generate an array of reactive oxidants and diffusible radical species. Although these leukocytes constitute an essential component of the effector limb of host defenses, they also are implicated in contributing to inflammatory tissue injury7.
The regulatory mechanisms of activation and degranulation of eosinophils occur in three stages. The first phase is the regulation of proliferation and differentiation in the bone marrow, which is consistent and occurring at a low level in the absence of immunological stimulation. When Th2 lymphocytes are activated to produce cytokines, such as IL-5, this increases the production of eosinophils in the bone marrow and promotes release of these cells into the circulation8. The second step is the migration of eosinophils from the circulation to various tissues. The migration of eosinophils depends on chemoattractants such as eotaxin, which is an eosinophil specific chemokine9. The third step comprises the release of granule proteins, which occurs when eosinophils become exposed to soluble mediators for example PAF10, immuncomplexes11 or solid particles12.
Four distinct populations of granule (primary granules, secondary or specific granules, small granules, lipid bodies) have been recognized in eosinophil13. The effector functions of eosinophils are of twofold. First, the release of highly toxic granule proteins stored preferentially in secondary granules. The primary granules contain lysophospholipase, associated with eosinophilic infiltrations14, and the secondary, or specific granules, contain major basic protein (MBP), eosinophils cationic protein (ECP), eosinophil peroxidase (EPO) and eosinophil protein X or eosinophil derived neurotoxin (EPX/EDN)15,16. The second effector function of the eosinophil is to produce and release inflammatory mediators such as PAF17, LTC417, IL-518 and eotaxin9, which amplify the inflammatory response by recruiting and activating leukocytes and epithelial cells.
Eosinophils develop from bone marrow (BM) progenitors, and interleukin-5 (IL-5) and eotaxin may act in expansion and mobilisation of eosinophils in asthma. The data suggest that IL-5 may act early in eosinophil development, and that eotaxin has the capacity to mobilise an eosinophil pool in asthma. The migration of eosinophils depends on chemoattractants such as eotaxin, which is an eosinophil specific chemokine9. Eotaxin represents the principal chemoattractant for the recruitment of the pulmonary eosinophils19. Eosinophils derive from CD34(+) bone marrow progenitor cells under the influence of haematopoietic growth factors, subsequently migrating to the airways under the cooperative influence of interleukin (IL)-5 and chemokines, including eotaxin4. There may be a critical role for Interleukin-13 in accumulation of intraepithelial eosinophils in chronic asthma, as well as in epithelial and subepithelial remodelling20.
Blood eosinophils are known to be an indirect marker of airway inflammation in asthma21. It is known since long that the total eosinophil count reflects asthmatic activity and is useful for regulating steroid dosage and for early detection of exacerbations22,23. Eosinophils are currently regarded as the effector cells responsible for much of the pathology of asthma24. Eosinophil-mediated damage to the respiratory epithelium is a major pathogenetic mechanism in asthma25. Atopy in general and asthma in particular are associated with blood and lung eosinophilia. The eosinophil mediates damage to the respiratory epithelium and is the prime effector cell in the pathophysiology of asthma16. Eosinophils play a major role in the onset and maintenance of bronchial inflammation and tissue injury in asthma26. Eosinophilia of lung and blood associated with injury to the mucociliary escalator and excessive shedding of bronchial epithelium are hallmarks of both allergic and nonallergic asthma16.
There is a 50-100 fold increase in the number of eosinophils relative to neutrophils in the bronchial mucosa in asthma. This is the result of the cumulative and sequential effects of increases in selective eosinophil versus neutrophil migration occurring at a number of stages in the life cycle of the eosinophil27. Blood eosinophil counts have for several decades been viewed as a valuable tool for indicating disease severity as they reflect the degree and extent of inflammation in the asthmatic lung28. Studies have reported a correlation between the number of blood eosinophils and the severity of asthma29,30. It has been reported that in the patients with asthma and pronounced eosinophilia the lung function of the patients is principally related to the number of circulating eosinophils29.
It has been determined that the degree of eosinophilia is proportional to the severity of asthma, as measured by clinical grading or pulmonary function22,29,31. Specific bronchial challenge, experimental and nonexperimental in sensitive individuals is associated with a short term rise in eosinophil count. Large numbers of bronchial submucosal and lumenal eosinophils are pathognomic in acute severe asthma refractory to therapy (status asthmaticus). Eosinophils are increased in bronchoalveolar lavage fluid from asthmatics compared to nonasthmatics29,32 as are subepithelial (and to a lesser extent deeper submucosal) eosinophils on bronchial biopsy31,33,34. In biopsy specimens from patients with symptomatic asthma, eosinophils are found in high numbers in the airway epithelium and in deeper bronchial tissues35. Two-thirds of people with mild to moderate asthma are reported to have increased sputum eosinophilia36.
Mechanism of exercise-induced asthma is undetermined however eosinophils have been suggested as playing a role in its occurrence. The influx of eosinophils to the airway in patients who develop exercise-induced asthma can be partially explained by the leukotrienes in the airways of those patients37. Eosinophils have been considered to play an important role in the pathogenesis of virus-induced asthma exacerbations38.
In the clinical management of patients with bronchial asthma blood eosinophilia is considered a risk factor, indicating deterioration and exacerbation39. However, patients with asthma may have a normal number of eosinophils in their blood40, similarly bronchial biopsy specimens from patients who have died of status asthmaticus do not always reveal eosinophils6 and histological studies performed after bronchoscopy in patients with asthma have not always found eosinophils in the bronchial mucosa41. Based on these results and the inconsistent findings of other investigators, we cannot depend on the eosinophilic count to monitor disease activity28-30,41.
PATHOPHYSIOLOGY
The role of the eosinophil in the pathophysiology of allergy and asthma has been the focus of intense interest during the last two decades. While the presence of eosinophils in humans with allergy and asthma is well established, the precise role of this cell in humans and in animal models is less clear. However, recent developments in research on many organ systems have provided novel insights into the possible underlying role of the eosinophil in both allergic and nonallergic inflammation42. The main role of the eosinophil seems to be a cytotoxic one, and it acts effectively both against host cells and parasites such as protozoa.
There are many potential mechanisms through which eosinophils may contribute to tissue injury and oxidative modification of biological targets in asthma7. Eosinophils may contribute to airway hyper responsiveness in asthma through the effects of eosinophil derived granular proteins in the bronchial epithelium43.
Eosinophils are a source of cytokines within the airways of asthmatic individuals that may exert an important immunoregulatory influence44. Eosinophils are a potential source of IL-12. Eosinophil-derived IL-12 may contribute and modulate the local allergic inflammatory responses44. There is evidence of increased expression of the CD4+ cell chemoattractant interleukin (IL)-16 in bronchial biopsies of atopic asthmatic subjects compared to normal controls. IL-16 immunoreactive cells were identified. T cells, eosinophils and mast cells comprised are the potential cellular sources of IL-16 in asthmatic airways45.
Like other leukocytes, eosinophils present in excessive numbers in inflamed tissues are removed by apoptosis. This phenomenon, also called 'programmed cell death', allows elimination of dangerous or redundant cells, thereby ensuring maintenance of tissue homeostasis. It has been suggested that a defect in eosinophil apoptosis would participate in the development and persistence of allergic airways inflammation in asthma26. A defect in apoptosis might contribute to the chronic tissue eosinophilia associated with asthma45. Apoptotic eosinophils are detectable in induced sputum of allergic patients. Apoptosis could be an important mechanism in the control of acute eosinophilic inflammation in patients with asthma exposed to the sensitizing antigens. It appears that the apoptotic mechanism could be less effective in controlling tissue eosinophilia in asthmatic patients with chronic eosinophilic inflammation46.
Eosinophil apoptosis, as well as the expression and function of various molecules determining this process, are closely regulated by various stimuli, including cytokines, lipid mediators and growth factors released by various cell types and by the eosinophil itself, as well as exogenous molecules, such as glucocorticoids26.,
There are increased numbers of circulating CD34(+) progenitor cells for eosinophils in patients with atopic asthma, with a further increase following allergen exposure or spontaneous worsening of asthma47. In addition, they suggest that in chronic asthma, IL-13 may be capable of signalling via a pathway that does not involve IL-4Ralpha20.
When activated, eosinophils undergo degranulation causing epithelial damage in the airway, desquamation and increased airway hypersensitivity6. Mechanisms leading to eosinophil degranulation are not well understood. At least three discrete processes have been defined that result in the release of granule contents from eosinophils: secretion, piecemeal degranulation and cytolysis (necrosis)13.
Eosinophil granules contain several proteins, which are secreted into the extracellular environment following stimulation. They include basic proteins, cytokines, chemokines, lipid mediators and oxygen radicals13. Charcot-Leyden crystal (CLC) protein (or lysophospholipase) makes up to 10% of the total protein in eosinophils. Its function is unknown, although it has been speculated that CLC may protect eosinophils from the toxic effects of lysophospholipids. The most distinguishing feature of the eosinophils is its high content of highly charged cationic proteins, eosinophil cationic protein (ECP), eosinophil protein X (EPX) and major basic protein (MBP). These proteins together with eosinophil peroxidase (EPO) have been implicated as final effector molecules in eosinophil-mediated tissue damage. All these proteins are shown to be highly toxic to cells of the respiratory epithelium6.
A number of studies have indicated that the assessment of eosinophil-derived proteins in various body fluids could be used for monitoring disease activity of asthma. Due to the relationship between levels of eosinophil proteins in serum/urine samples and lung function, as well as significant concentration differences between symptomatic and asymptomatic asthmatics, the assessment of eosinophil proteins in serum or urine samples appear to be more appropriate in monitoring disease activity.
In addition, recent reports have provided convincing evidence that bronchial inflammation and elevated levels of eosinophil granule associated proteins are even present in the bronchoalveolar lavage fluid of patients whose asthma is stable, suggesting that ongoing recruitment and activation of inflammatory cells may also be occurring in asymptomatic asthmatic individuals31. The potency of the eosinophil to release these mediators can be greatly enhanced by eosinophil mobilizing and activating factors such as granulocyte macrophage colony stimulating factor, interleukins 3 and 5, platelet-activating factor, and perhaps leukotriene B440.
Major basic protein (MBP) is regarded as the most important out of these. This 31 kD protein is highly basic, and negatively charged in solution, and exhibits potent cytotoxic activity. Eosinophilic cationic protein (ECP) and eosinophil-derived neurotoxin (18-21 kD) are highly homologous proteins and exhibit ribonuclease activity. Both lead to exfoliation of respiratory epithelium. Both are highly basic. ECP also stimulates mast cell degranulation, inhibits T-cell activity, and shortens the coagulation time. Eosinophil peroxidase is cytotoxic in itself, but more so in the presence of hydrogen peroxide and halides. It too can cause mast cell degranulation 6.
ECP AND ASTHMA
Eosinophil cationic protein (ECP) is an eosinophil granule protein which is highly cytotoxic and is released by activated eosinophils10. Measurement of serum ECP is helpful in determining asthma activity and deciding the use of anti-asthma drugs48. There is a correlation of ECP and the eotaxin level in asthma patients49. Measuring serum ECP levels has the advantage over eosinophilic count in that it reflects not only the number of eosinophils but also their degree of activation and is therefore a better inflammatory marker50. In asthmatics, sECP is associated with PEF variability and symptom severity51. Although the ECP values are significantly more elevated in atopic than in non-atopic asthma, elevated serum ECP are not specific for atopic asthma52.
Measurement of serum ECP is helpful in determining asthma activity and deciding the use of anti-asthma drugs48. ECP has also been suggested to be a helpful indicator of persistent infantile wheeze53. There is a significant correlation between sECP level and blood eosinophil count as well as between sECP level and serum tIgE. There is a significant correlation of asthma attack frequency with sECP level. Serum ECP level seems to reflect the allergy activity level, especially two weeks prior to and after the measurement54.
ECP levels, however, may not be elevated in some asthmatic patients, even when they are symptomatic. It may not be useful in patients who are older, have longer disease duration or possibly have thicker airway walls55.
Levels of salivary ECP are elevated in patients with asthma and associated with presumed activity of disease as recorded by alteration of taken dose of inhaled corticosteroid24. The serum level of ECP seems to be a good clinical marker of monitoring the disease. Serum levels of ECP in the patients with asthma during attack were significantly higher than those of the controls and stayed high up to the 7th day1. Serum ECP may be used to predict a response to corticosteroid therapy in adult patients with asthma56.
MBP AND ASTHMA
Eosinophil granule major basic protein (MBP) is present at sites of epithelial damage in cases of asthma; this protein is toxic to respiratory epithelium and its levels are raised in the sputum of patients with asthma. MBP in the eosinophil granule is released into respiratory tissue of patients with severe asthma and that it is associated with tissue damage. In vitro, MBP causes shedding of respiratory epithelium. In vivo, levels of MBP in the sputum of asthmatics reflect severity of current disease, while immunofluorescent studies show that MBP is present in the respiratory epithelial defects seen in asthmatics’ bronchi57. In vitro, the eosinophil granule major basic protein (MBP) is toxic to the MBP-mediated damage to the respiratory epithelium consists of desquamation and frank destruction of ciliated cells16. Increased sputum MBP concentration is a good marker for asthma, and patients treated for acute asthma have high levels of MBP in their sputa, which decrease after treatment. In the lungs of patients who had died of asthma, MBP has been localized outside of the eosinophil in association with damage to the epithelium16.
EDN/EPX AND ASTHMA
Eosinophil-derived neurotoxin (EDN), also called eosinophil protein X (EPX), has been suggested to be a useful marker of eosinophilic inflammation. EDN levels in serum, plasma and urine from the patients significantly correlated with the number of peripheral blood eosinophils, but not total serum IgE levels58. Eosinophil protein X (EPX) has been used to assess eosinophil activity and monitor inflammation in bronchial asthma59.
Recently there has been a lot of interest in urinary EPX, which can be obtained easily and that may be helpful in diagnosing both asthma and atopy in children. However, there is a great overlap between controls and symptomatics60. Urinary EPX is a useful marker of airway inflammation and can be helpful in guiding asthma management3,59.
It has been suggested that urinary eosinophil protein X (U-EPX) can be used to monitor bronchial inflammation in childhood asthma. It may reflect differences in eosinophil involvement and activation between children with atopic and non-atopic asthma61. A statistically significant correlation was found between urinary concentration of eosinophil protein X (U-EPX) concentrations and the severity of attacks in asthmatic children60.
Eosinophil blood counts and eosinophil cation protein (ECP) serum level are often used as markers of clinical monitoring of the disease activity25. Correct prediction of clinical response in an individual patient, however, remains poor with our currently used clinical and inflammatory parameters62. The proportion of eosinophils in sputum is a more sensitive marker of asthmatic airway inflammation than the proportion of eosinophils in blood63. Eosinophil count and/or ECP and EPO levels can be used to estimate the short-term risk of deterioration and the need for corticosteroid treatment in cases of mild and moderate allergic asthma64. The development of commercial kits for other markers is still in the process.
Treatment decisions in asthma are based on assessments of symptoms and simple measures of lung function, which do not relate closely to underlying eosinophilic airway inflammation65. Corticosteroids are considered to be one of the most effective medicine for asthma by suppressing airway inflammation and Prednisolone is considered to be effective medicine for asthma by suppressing eosinophil activation through IL-566. A treatment strategy directed at normalisation of the induced sputum eosinophil count reduces asthma exacerbations and admissions without the need for additional anti-inflammatory treatment65.
FUTURE RESEARCH
The above review indicates that the relationship of eosinophils and asthma is very important for diagnosis and management of asthma. There is a need to work on all the aspects of this relationship that includes finding correlation of symptoms of asthma with markers of eosinophil activity, identifying the substances attracting, activating or developing eosinophils and developing drugs to neutralize these substances. Works are now underway to develop asthma therapy leading to inhibition of eosinophil priming of cytotoxic mechanisms in vivo67. For the development of more effective anti-asthma drugs it, therefore, seems relevant to unravel and interfere with the steps of eosinophil activation40. There are now attempts to inhibit eosinophil differentiation at bone marrow level68.
Eosinophilic inflammation is a feature of asthma. However, serological markers to indicate eosinophil activation in this process are not fully defined56. Markers of airway inflammation are needed for prediction of asthma deterioration and evaluation of disease severity64. It is, therefore, important to find a marker of disease activity, ideally one that is simple to measure, reliable and inexpensive. As yet no such marker has been found for asthma. Therefore, there is a need for assessing different eosinophil products to develop a serological marker of airway inflammatory activity in asthma.
1. Kokuludag A, Sin A, Terzioglu E, Saydam G, Sebik F. Elevation of serum eosinophil cationic protein, soluble tumor necrosis factor receptors and soluble intercellular adhesion molecule-1 levels in acute bronchial asthma. J Investig Allergol Clin Immunol 2002;12(3):211-4
2. Norzila MZ, Fakes K, Henry RL, Simpson J, Gibson PG. Interleukin-8 secretion and neutrophil recruitment accompanies induced sputum eosinophil activation in children with acute asthma. Am J Respir Crit Care Med 2000 Mar;161(3 Pt 1):769-74
3. Labbe A, Aublet-Cuvelier B, Jouaville L, Beaugeon G, Fiani L, Petit I, Ouchchane L, Doly M. Prospective longitudinal study of urinary eosinophil protein X in children with asthma and chronic cough. Pediatr Pulmonol 2001 May;31(5):354-62
4. Stirling RG, van Rensen EL, Barnes PJ, Chung KF. Interleukin-5 induces CD34(+) eosinophil progenitor mobilization and eosinophil CCR3 expression in asthma. Am J Respir Crit Care Med 2001 Oct 15;164(8 Pt 1):1403-9
5. Coyle AJ, Gutierrez-Ramos JC. Eosinophils in asthma. In: Celleular mechanisms in airway infl ammation. Eds. CP Page, KH Banner, D Spina. Birkhäuser Verlag, ISBN 3-7643-5852-1, 2000. 147-158.
6. Gleich GJ (1990): The eosinophil and bronchial asthma: current understanding. J Allergy Clin Immunol 85: 422-436.
7. Mitra SN, Slungaard A, Hazen SL. Role of eosinophil peroxidase in the origins of protein oxidation in asthma. Redox Rep 2000;5(4):215-24
8. Clutterbuck EJ, Sanderson CJ. Human eosinophil hematopoiesis studied in vitro by means of murine eosinophil differentiation factor (IL5): production of functionally active eosinophils from normal human bone marrow. Blood 1988;71:646-651.
9. Rothenberg ME. Eotaxin. An essential mediator of eosinophil trafficking into mucosal tissues. Am J Respir Cell Mol Biol. 1999;21:291-295.
10. Venge P. Serum measurement of eosinophil cationic protein (ECP) in bronchial asthma. Clinical and experimental allergy, 1993, 23 (suppl. 2):3-7.
11. Takenaka T, Okuda M, Kawabori S, Kubo K. Extracellular release of peroxidase from eosinophils by interaction with immune complexes. Clin & Exp Immunol 1977;28:56-60.
12. Winqvist I, Olofsson T, Olsson I. Mechanisms for eosinophil degranulation; release of the eosinophil cationic protein. Immunology 1984;51:1-8.
13. Giembycz MA, Lindsay MA. Pharmacology of the eosinophil. Pharmacol Rev1999;51:213-339.
14. Dvorak AM, Letourneau L, Login GR, Weller PF, Ackerman SJ. Ultrastructural localization of the Charcot-Leyden crystal protein (lysophospholipase) to a distinct crystalloid-free granule population in mature human eosinophils. Blood 1988;72:150- 158.
15. Spry C. Eosinophils: a comperhensive review and guide to the scientific and medical literature. Oxford University press Ltd., Oxford, UK. 1998.
16. Frigas E, Motojima S, Gleich GJ. The eosinophilic injury to the mucosa of the airways in the pathogenesis of bronchial asthma. Eur Resp J 1991;13(S):123-135.
17. Blom M, Tool AT, Wever PC, Wolbink GJ, Brouwer MC, Calafat J, Egesten A, Knol EF, Hack CE, Roos D, Verhoeven AJ. Human eosinophils express, relative to other circulating leukocytes, large amounts of secretory 14-kD phospholipase A2. Blood 1998;91:3037-3043.
18. Lundahl J, Halldén G, Hed J. Differences in intracellular pool and receptor-dependent mobilization of the adhesion-promoting glycoprotein Mac-1 between eosinophils and neutrophils. J Leukoc Biol 1993;53:336-341.
19. Kim J, Merry AC, Nemzek JA, Bolgos GL, Siddiqui J, Remick DG. Eotaxin represents the principal eosinophil chemoattractant in a novel murine asthma model induced by house dust containing cockroach allergens. J Immunol 2001 Sep 1;167(5):2808-15
20. Kumar RK, Herbert C, Yang M, Koskinen AM, McKenzie AN, Foster PS. Role of interleukin-13 in eosinophil accumulation and airway remodelling in a mouse model of chronic asthma. Clin Exp Allergy 2002 Jul;32(7):1104-11
21. Koh YI, Choi S. Blood eosinophil counts for the prediction of the severity of exercise-induced bronchospasm in asthma. 14: Respir Med 2002 Feb;96(2):120-5
22. Horn BR, Robin ED, Theodore J, Van Kessel A. Total eosinophil counts in the management of bronchial asthma. N Engl J Med 1975 May 29;292(22):1152-5
23. Gupta S, Frenkel R, Rosenstein M, Grieco MH. Lymphocyte subpopulations, serum IgE and total eosinophil counts in patients with bronchial asthma. Clin Exp Immunol 1975 Dec;22(3):438-45
24. Schmekel B, Ahlner J, Malmstrom M, Venge P. Eosinophil cationic protein (ECP) in saliva: a new marker of disease activity in bronchial asthma. Respir Med 2001 Aug;95(8):670-5
25. Stelmach I, Grzelewski T, Stelmach W, Majak P, Jerzynska J, Gorski P, Kuna P. Effect of triamcinolone acetonide, montelukast, nedocromil sodium and formoterol on eosinophil blood counts, ECP serum levels and clinical progression of asthma in children]. Pol Merkuriusz Lek 2002 Mar;12(69):208-13
26. Druilhe A, Letuve S, Pretolani M. Eosinophil apoptosis in asthma. Pathol Biol (Paris) 2000 Jul;48(6):566-73
27. Wardlaw AJ. Eosinophil trafficking in asthma. Clin Med 2001 May-Jun;1(3):214-8
28. Dahl R. Monitoring bronchial asthma in the blood. Allergy, 1993, 48:77-80.
29. Griffin E, Hakansson L, Formgren H, Jorgensen K, Peterson C, Venge P. Blood eosinophil number and activity in relation to lung function in patients with asthma and with eosinophilia. J Allergy Clin Immunol 1991 Feb;87(2):548-57
30.
Burrows Bl. Epidemiologic observations on eosinophilia and its relation
to respiratory disorders. American review of respiratory disease, 1980,
122(5):709-19.
31. Bousquet J, Chanez P, Lacoste JY, Barnéon G, Ghavanian N, Enander I et al. Eosinophilic inflammation in asthma. New Engl J Med 1990;323: 1033-1039.
32. DeMonchy JGR, Kauffman HF, Venge F, Koëter GH, Jansen HM, Sluiter HJ, De Vries K (1985): Bronchoalveolar eosinophilia during allergeninduced late asthmatic reactions. Rev Respir Dis 131: 373-376.
33. Djukanovic R, Wilson JW, Britten KM, Wilson SJ, Walls AF, Roche WR et al. Quantitation of mast cells and eosinophils in the bronchial mucosa of symptomatic atopic asthmatics and healthy control subjects using immunohistochemistry. Am Rev Respir Dis 1990;142: 863-871.
34. Beasley R, Roché WR, Roberts JA, Holgate ST (1989): Cellular events in the bronchi in mild asthma and after bronchial provocation. Am Rev Respir Dis 1989;139: 806-817.
35. Laitinen LA, Laitinen A, Haahtela T. Airway mucosal inflammation even in patients with newly diagnosed asthma. Am Rev Respir Dis 1993;147:697-704.
36. Louis R, Sele J, Henket M, Cataldo D, Bettiol J, Seiden L, Bartsch P. Sputum eosinophil count in a large population of patients with mild to moderate steroid-naive asthma: distribution and relationship with methacholine bronchial hyperresponsiveness. 5: Allergy 2002 Oct;57(10):907-12
37. Kivity S, Argaman A, Onn A, Shwartz Y, Man A, Greif J, Fireman E. Eosinophil influx into the airways in patients with exercise-induced asthma. Respir Med 2000 Dec;94(12):1200-5
38. Zhao J, Takamura M, Yamaoka A, Odajima Y, Iikura Y. Altered eosinophil levels as a result of viral infection in asthma exacerbation in childhood. 11: Pediatr Allergy Immunol 2002 Feb;13(1):47-50
40. Bruijnzeel PL. Contribution of eosinophil-derived mediators in asthma. Int Arch Allergy Appl Immunol 1989;90 Suppl 1:57-63
41. Lundgren R et al. Morphological studies of bronchial mucosal biopsies from asthmatics before and after ten years of treatment with inhaled steroids. European respiratory journal, 1988, 1(10):883-9.
42. Adamko D, Lacy P, Moqbel R. Mechanisms of Eosinophil Recruitment and Activation. Current Allergy and Asthma Reports 2002 2:107-116.
43. Onadeko BO, Khadadah ME, Mustafa HT, Ezeamuzie CI, Marouf R, Khamis A, et al. Serum eosinophil cationic protein as a predictor of disease activity in acute and chronic asthma. East Afr Med J 1999 Sep;76(9):524-9
44. Nutku E, Gounni AS, Olivenstein R, Hamid Q. Evidence for expression of eosinophil-associated IL-12 messenger RNA and immunoreactivity in bronchial asthma. J Allergy Clin Immunol 2000 Aug;106(2):288-92
45. Kankaanranta H, Lindsay MA, Giembycz MA, Zhang X, Moilanen E, Barnes PJ et al. Asthma attack severity and urinary concentration of Eosinophil X protein in children. Allergol Immunopathol (Madr) 2002 Jul-Aug;30(4):225-31
46. Foresi A, Teodoro C, Leone C, Pelucchi A, D'Ippolito R, Chetta A, Olivieri D. Eosinophil apoptosis in induced sputum from patients with seasonal allergic rhinitis and with asymptomatic and symptomatic asthma. Ann Allergy Asthma Immunol 2000 Apr;84(4):411-6
47. Kuo HP, Wang CH, Lin HC, Hwang KS, Liu SL, Chung KF. Interleukin-5 in growth and differentiation of blood eosinophil progenitors in asthma: effect of glucocorticoids. Br J Pharmacol 2001 Dec;134(7):1539-47
48. Badr-el-Din OM, el-Sawy IH, el-Azzouni OE, Badr-el-Din MM, Salem AM. Eosinophil cationic protein as a serological marker of disease activity in childhood bronchial asthma. East Mediterr Health J 1999 Jul;5(4):664-75
49. Jahnz-Ro yk K, Plusa T, Mierzejewska J. Eotaxin in serum of patients with asthma or chronic obstructive pulmonary disease: relationship with eosinophil cationic protein and lung function. Mediators Inflamm 2000;9(3-4):175-9
50. Koller DY, Halmerbauer G, Frischer T, Roithner B. Assessment of eosinophil granule proteins in various body fluids: is there a relation to clinical variables in childhood asthma? Clin Exp Allergy 1999 Jun;29(6):786-93
51. Perfetti L, Galdi E, Brame B, Speciale L, Moscato G. Serum eosinophil cationic protein (sECP) in subjects with a history of asthma symptoms with or without rhinitis. Allergy 1999 Sep;54(9):962-7
52. Melbostad E, Venge P, Danielsen TA, Kjuus H. Serum eosinophil cationic protein in asthma of farmers. Scand J Clin Lab Invest 2000 Apr;60(2):111-8
53. Marguet C, Dean TP, Basuyau JP, Warner JO. Eosinophil cationic protein and interleukin-8 levels in bronchial lavage fluid from children with asthma and infantile wheeze. Pediatr Allergy Immunol 2001 Feb;12(1):27-33
54. Ishigaki N, Masuhara C, Sakamaki K, Ishikawa Y, Ohta K, Koike R, Mikuni K, Haruna H, Awa S. Relation between serum eosinophil cationic protein (ECP) level and asthma attack in children. Arerugi 2000 Nov;49(11):1093-103
55. Matsumoto H, Niimi A, Minakuchi M, Izumi T. Serum eosinophil cationic protein levels measured during exacerbation of asthma: characteristics of patients with low titres. Clin Exp Allergy 2001 Apr;31(4):637-43
56. Sorkness C, McGill K, Busse WW. Evaluation of serum eosinophil cationic protein as a predictive marker for asthma exacerbation in patients with persistent disease. Clin Exp Allergy 2002 Sep;32(9):1355-9
57. Filley WV, Holley KE, Kephart GM, Gleich GJ.Identification by immunofluorescence of eosinophil granule major basic protein in lung tissues of patients with bronchial asthma. Lancet 1982 Jul 3;2(8288):11-6
58. Morioka J, Kurosawa M, Inamura H, Nakagami R, Mizushima Y, Chihara J, et al. Development of a novel enzyme-linked immunosorbent assay for blood and urinary eosinophil-derived neurotoxin: a preliminary study in patients with bronchial asthma. Int Arch Allergy Immunol 2000 May;122(1):49-57
59. Severien C, Artlich A, Jonas S, Becher G. Urinary excretion of leukotriene E4 and eosinophil protein X in children with atopic asthma. Eur Respir J 2000 Oct;16(4):588-92
60. Studnicka M. Urinary eosinophil protein X in children: the relationship to asthma and atopy and normal values. Allergy 2000 Jul;55(7):647-52
61. Oymar K. High levels of urinary eosinophil protein X in young asthmatic children predict persistent atopic asthma. Pediatr Allergy Immunol 2001 Dec;12(6):312-7
62. Meijer RJ, Postma DS, Kauffman HF, Arends LR, Koeter GH, Kerstjens HA. Accuracy of eosinophils and eosinophil cationic protein to predict steroid improvement in asthma. Clin Exp Allergy 2002 Jul;32(7):1096-103
63. Pizzichini E, Pizzichini MMM, Efthimiadis A, Dolovich J, Hargreave FE. Measuring airway infl ammation in asthma: Eosinophils and eosinophilic cationic protein in induced sputum compared with peripheral blood. J Allergy Clin Immunol 1997;99:539-544.
64. Lonnkvist K, Hellman C, Lundahl J, Hallden G, Hedlin G. Eosinophil markers in blood, serum, and urine for monitoring the clinical course in childhood asthma: impact of budesonide treatment and withdrawal. J Allergy Clin Immunol 2001 May;107(5):812-7
65. Green RH, Brightling CE, McKenna S, Hargadon B, Parker D, Bradding P, Wardlaw AJ, Pavord ID. Asthma exacerbations and sputum eosinophil counts: a randomised controlled trial. Lancet 2002 Nov 30;360(9347):1715-21
66. Jang AS, Choi IS, Koh YI, Jeong TK, Lee KY, Kim YS, Lee JU, Park CS. Effects of prednisolone on eosinophils, IL-5, eosinophil cationic protein, EG2+ eosinophils, and nitric oxide metabolites in the sputum of patients with exacerbated asthma. J Korean Med Sci 2000 Oct;15(5):521-8
67. Grutters JC, Brinkman L, Aslander MM, van den Bosch JM, Koenderman L, Lammers JW. Asthma therapy modulates priming-associated blood eosinophil responsiveness in allergic asthmatics. Eur Respir J 1999 Oct;14(4):915-22
68. Rais M, Wild JS, Choudhury BK, Alam R, Stafford S, Dharajiya N, Sur S. Interleukin-12 inhibits eosinophil differentiation from bone marrow stem cells in an interferon-gamma-dependent manner in a mouse model of asthma. Clin Exp Allergy 2002 Apr;32(4):627-32
Address for Correspondence:
Dr. Ahmed Badar, Email: Badar@ayubmed.edu.pk