- Email: [email protected]
Eosinophils and basophils in allergic airway inflammation
Busse et
al.
J ALLERGY CLIN IMMUNOL
DECEMBER1994
26. Talbot SF, Atkins PC, Valenzano M, et al. Correlations of in vivo mediator release with late cutaneous allergic response in humans. I. Kinetics of histamine release. J ALLERGYCLIN IMMUNOL1984;74:819-26. 27. Shalit M, Schwartz LB, Golzar N, et al. Release of histamine and tryptase in vivo after prolonged cutaneous challenge with allergen in humans. J Immunol 1988;141: 821-6. 28. Talbot SF, Atkins PC, Goetzl EJ, et al. Accumulation of leukotriene C4 and histamine in human allergic skin reactions. J Clin Invest 1985;76:650-6. 29. Shalit M, Valone FH, Atkins PC, et al. Late appearance of phospholipid platelet-activating factor and leukotriene B4 in human skin after repeated antigen challenge. J ALLERCY CLIN IMMUNOL1989;83:691-6. 30. Tamura N, Agrawal DK, Suliaman, et al. Effects of platelet activating factor on the chemotaxis of normodense eosinophils from normal subjects. Biochem Biophys Res Commun 1987;142:638-44. 31. Chilton FH, Connell TR. 1-0-Alkyl-2-arachidonoyl-snglycero-3-phosphocholine: a common source of plateletactivating factor and arachidonate in human polymorphonuclear leukocytes. J Biol Chem 1984;259:12014-9. 32. Bochner BS, Charlesworth EN, Kagey-Sobotka A, et al. Interleukin-1 is released at sites of human cutaneous allergic reactions. J ALLERGYCLINIMMUNOL1990;86:830-9. 33. Schleimer RP, Rutledge BK. Cultured human vascular endothelial cells acquire adhesiveness for neutrophils after stimulation with interleukin 1, endotoxin, and tumorpromoting phorbol esters. J Immunol 1986;135:649-54. 34. Beviliaqua MP, Pober JS, Mendrick, et al. Identification of an inducible endothelial-leukocyte adhesion molecule. Proc Natl Acad Sci USA 1987;84:9238-42. 35. Subramanian N, Bray MA. Interleukin 1 releases histamine from human basophils and mast cells in vitro. J Immunol 1987;138:271-5. 36. Haak-Frendscho M, Dinarello CA, Kaplin AP, et al. Recombinant human interleukin 1 beta causes histamine release from human basophils. J ALLEROVCLIN IMMUNOL 1988;82:218-23. 37. Lee CE, Neuland ME, Teaford HG, et al. Interleukin-6 is released in the cutaneous response to allergen challenge in atopic individuals. J ALLERGYCLIN IMMUNOL1992;89: 1010-20.
38. Moller A, Lippert U, Lessmann D, et al. Human mast cells produce IL-8. J Immunol 1993;151:3261-6. 39. Bradding P, Feather IH, Wilson S, et al. Immunolocalization of cytokines in the nasal mucosa of normal and perennial rhinitic subjects: the mast cell as a source of IL-4, IL-5, and IL-6 in human allergic mucosal inflammation. J Immunol 1993;151:3853-65. 40. Morris FM, Whisman BA, Charlesworth EN. Measurement of TNF/alpha, GM-CSF, and IL-4 in the cutaneous late-phase response to antigen [Abstract]. J ALLERGYCLIN IMMUNOL1994;93:261. 41. Massey W, Friedman B, Kato M, et al. Appearance of granulocytes-macrophage colony stimulating factor activity at allergen-challenged cutaneous late-phase reaction sites. J Immunol 1993;150:1084-92. 42. Varney VA, Hamid QA, Gaga M, et al. Influence of grass pollen immunotherapy on cellular infiltration and cytokine mRNA expression during allergen-induced late-phase cutaneous responses. J Clin Invest 1993;92:644-51. 43. Pipkorn U, Proud D, Lichtenstein LM, et al. Effect of short-term glucocorticoid treatment on human nasal mediator release after antigen challenge. J Clin Invest 1987; 80:957-61. 44. Charlesworth EN, Kagey-Sobotka A, Schleimer RP, et al. Prednisone inhibits the appearance of inflammatory mediators and the influx of eosinophils and basophils associated with the cutaneous late-phase response to allergen. J Immunol 1991;146:671-6. 45. Schleimer RP, Lichtenstein LM, Gillespie E. Inhibition of basophil histamine release by anti-inflammatory steroids. Nature 1981;292:454-5. 46. Fadel R, Herpin-Richard N, Rihoux JP, Henolg E. Inhibitory effect of cetirizine 2 HC1 on eosinophilia in vivo. Clin Allergy 1987;17:373-6. 47. Charlesworth EN, Massey WA, Kagey-Sobotka A, Norman PS, Lichtenstein LM. Effect of H1 receptor blockade on the early and late response to cutaneous allergen challenge. J Pharmacol Exp Ther 1992;262:964-70. 48. Alam R, Dejarnatt A, Stafford S, et al. Selective inhibition of the cutaneous late but not immediate allergic response to antigens by misoprostol, a PGE analog. Am Rev Respir Dis 1993;148:1066-70.
Eosinophils and basophils in allergic airway inflammation William W. Busse, MD, Julie B. Sedgwick, Nizar N. Jarjour, and William J. Calhoun Madison, Wis.
From the Section of Allergy and Clinical Immunology, Department of Medicine, University of Wisconsin. J ALLERGYCUN IMMVNOL1994;94:1250-4. Copyright © 1994 by Mosby-Year Book, Inc. 0091-6749/94/$3.00 + 0 1/0/59927
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A i r w a y i n f l a m m a t i o n h a s b e e n a c c e p t e d as a feature of asthma and other allergic inflammatory d i s e a s e s . T h e m a n y cells p a r t i c i p a t i n g in this p r o c e s s i n c l u d e m a s t cells, b a s o p h i l s , e o s i n o p h i l s , a n d l y m p h o c y t e s . O f t h e s e cells t h e m a s t cell, e o s i n o -
J ALLERGYCLIN IMMUNOL VOLUME 94, NUMBER6, PART 2
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Abbreviations used
BAL: Bronchoalveolar lavage FEVI: Forced expiratory volume in 1 second PC2o: Provocative concentration needed to decrease FEV 1 by 20% PGD2: Prostaglandin D2 SBP: Segmental bronchoprovocation
±
=x 0 24 E o~ 18 e-
phil, and lymphocyte have attracted greatest attention. Further, these cells are easiest to study in the in vivo setting because they possess measurable mediators, characteristic morphology, and distinct surface receptors, and because they can be isolated in sufficient number for in vitro study. In contrast, the basophil is a less dominant cell both in circulation and airway secretions. Thus efforts to define its unique or interactive contribution to allergic inflammation have been more difficult and insights obtained remain less specific. The following article discusses evidence that basophil function may have a correlation with markers of asthma severity, participate in the late allergic airway reaction, and predict eosinophilic participation in the airway's response to allergen. BASOPHIL HISTAMINE RELEASE AND AIRWAY RESPONSIVENESS Many mechanisms determine basophil histamine release, including membrane-bound IgE and cytokine exposure. 1 However, the absolute concentration of membrane-bound IgE does not correlate with the degree of basophil histamine released after an IgE-dependent stimulus2; consequently it has been proposed that basophil secretion may be determined by an intrinsic cellular property defined as "releasability. ''3 A number of features of basophil releasability have already been determined. Furthermore, abnormalities in basophil release of histamine have been noted in atopic dermatitis, 4' 5 allergic bronchopulmonary aspergillus, 6 food hypersensitivity, 7 allergic rhinitis, 8" 9 chronic urticaria, 1° and asthma. 1144 Although these associations show a potential relationship to allergic diseases, they do not identify the mechanism of enhanced releasability. As will be discussed, basophil histamine release may participate in the process airway inflammation and be linked to or contribute to airway responsiveness. In previous work we expanded on earlier observations and evaluated basophil histamine release in subjects with asthma. 14 The premise of
LIJ 12 03 I-O. 6 cr I---<
SALINE
LOW
MED
HIGH
BAL SEGMENT (AG DOSE) FIG. 1. Comparison of leukocyte histamine release to anti-lgE (10 to 1000 ng/ml) in six normal subjects, five nonallergic patients with asthma and seven allergic patients with asthma. Values represent mean (-+SEM) percentage histamine release. Asterisks denote degree of significance when asthma groups are compared with normal group. Spontaneous histamine release was subtracted from simulated release values. (From Gaddy JN, Busse WW. A m Rev Respir Dis 1986;134:969-74.)
this study was to determine whether any relationship existed between IgE-dependent basophil histamine releasability and a pathognomonic feature of asthma, bronchial responsiveness. In this study 16 patients with asthma were selected for examination. Of these, nine demonstrated immediate skin test reactivity to common inhalant allergens; the other seven had no evidence of skin test reactivity and were considered to have nonallergic asthma. In addition, seven control subjects were identified who were age matched but who had no evidence of allergic disease or asthma. With the use of anti-IgE as the basophil-secretagogue, both allergic and nonallergic asthmatic subjects released histamine, as did the nonallergic control group (Fig. 1). At all concentrations of anti-IgE used, basophils from both asthma populations, allergic and nonallergic, released more histamine than did the control population. These data indicate that basophil histamine release may not be dictated alone by the concentration of surface-bound IgE. Rather, other factors (i.e., basophil releasability) were likely to be important. Furthermore, enhanced basophil releasability was a feature of asthma and not determined by the presence of allergic disease. To extend these observations we evaluated the relationship of basophil histamine release to a
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FIG. 2. Correlation between anti-lgE-dependent leukocyte histamine release and PC2ofor aerosol histamine was evaluated in 12 subjects with asthma (eight allergic, four nonallergic). Values represent percentage of histamine release at anti-lgE concentration of 30 ng/ml versus log e PC2o for each subject. There was significant (p < 0.05) inverse correlation as determined by linear regression analysis and Pearson's rank correlation between percentage of histamine release and log e PC2o. (From Gaddy JN, Busse WW. Am Rev Respir Dis 134:969-74.)
marker of asthma: airway responsiveness to histamine measured by the provocative concentration required to decrease the forced expiratory volume in 1 second (FEV0 by 20% (PC20). A significant inverse correlation was noted between PC20 to histamine and three concentrations of anti-IgE: 30.0 ng/ml (r = -0.753), 100 ng/ml (r = -0.678), and 300 ng/ml (r = -0.671) (Fig. 2). These data indicate that the greater the level of basophil histamine release to anti-IgE, the more responsive to inhaled histamine was the patient with asthma. Although these data do not indicate that basophil histamine releasability causes bronchial hyperresponsiveness, it raised the question whether this feature of the basophil may be a reflection of similar abnormalities in the airway of patients with asthma. It is intriguing to consider the possibility that basophil releasability and airway hyperresponsiveness share a same intrinsic abnormality in signal transduction, and this abnormality may account for releasability and responsiveness. EVIDENCE THAT BASOPHILS PARTICIPATE IN THE LATE AIRWAY ALLERGIC REACTION
Mast cells and baosphils are cellular sources of airway histamine. The mast cell makes the dominant contribution to the airway histamine "pool." Casale et al. 15 and other investigators 16 found increased bronchoalveolar lavage (BAL) concentrations of histamine in patients with allergic
asthma when compared with those with allergic rhinitis and with normal subjects. Although both patients with allergic asthma and those with rhinitis had increased concentrations of mast cells in the lavage fluid compared with normal subjects, there was no correlation between BAL mast cell members and lavage fluid concentration of histamine. No mention was made of the number of basophils in the airway lavage fluid. In addition, BAL histamine concentrations were greater in those subjects with increased airway responsiveness. These data raise the possibility that ongoing mediator release and presumably other factors of allergic inflammation contribute to altered airway function in asthma, and the airway fluid concentration of histamine is a marker of this process. The relative contribution of mast cells and basophils to this process was not clarified. Evidence for basophil participation in the allergic inflammatory process first came from work by Naclerio et al. 17 Antigen challenge of the nose elicits an immediate response of sneezing and mediator release, including histamine, kinins, tosyl-arginine methyl esterases, and prostaglandin Dz (PGD2). In contrast, the late response to antigen is characterized by an increase in histamine but not in PGD2. Because PGD2 is derived from mast cells alone, the late-phase rise in histamine in nasal fluids was attributed to basophil recruitment to the nose and activation. This conclusion has been substantiated by cell staining. 18 Similar findings have been noted in the lower part of the airway. Liu et a l . 19 conducted localized antigen challenge of airway segments by bronchoscopy. Lavage fluid obtained 5 minutes after antigen challenge had increased levels of histamine, PGD2, and kinins. A similar profile of mediator release increase was noted 19 hours after the antigen challenge; these data do not indicate which mediator-releasing cells contributed to the "late" responses to antigen. However, Liu et al. were able to show an increase in alcian bluestaining cells in the late-phase lavage fluid. Furthermore, the predominant alcian blue-staining cell in this latter category was the basophil. These data support the concept that basophils are recruited to the airway after antigen challenge and release mediators in the late phase of the allergic reaction. We have also used segmental bronchoprovocation (SBP) as a method to more clearly define the airway allergic response to antigen. 2° In our protocol isolated airways are challenged with saline
J ALLERGY CLIN IMMUNOL VOLUME 94, NUMBER 6, PART 2
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90
@ m 0
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l 60
Non-AllergicAsthma, n=5 Allergic Asthmo,n=7 p
Q~
I=
45
-io
g SPONTANEOUS
I0
30 [Anti-IgE]
IO0
~500
I000
(ng/ml)
FIG. 3. Mediator release after antigen challenge. BAL fluids were assayed immediately (open bars, 12 minutes) and late (hatched bars, 48 hours) for histamine (top) and tryptase (bottom).
solution or relevant antigen. Lavage is performed immediately after antigen challenge (N 5 minutes) and again 48 hours later. There was a marked increase in histamine and tryptase immediately after antigen challenge (Fig. 3). The increase in these mast cell mediators was proportional to the concentration of antigen delivered to the airway. In contrast, when BAL fluids were obtained 48 hours after antigen challenge, tryptase values were similar to baseline concentrations of this mediator. Histamine concentrations in lavage fluid, however, showed a different pattern. Although the concentrations of histamine obtained immediately after antigen challenge were markedly elevated, lavage fluid histamine was still detectable at 48 hours after antigen challenge and concentrations were significantly elevated over baseline values. Because tryptase was not detected in the 48-hour postchallenge lavage sample, we concluded that a probable cell source of histamine was the basophil. This conclusion has to remain presumptive because we did not detect basophils in cells from the late lavage fluid. Nonetheless, in light of data from Johns Hopkins investigations that basophils are a component of the late response to antigen, we speculate that these late-phase increases in histamine reflect a basophil contribution to the airway allergic response. PERIPHERAL BLOOD BASOPHIL HISTAMINE RELEASE AND EOSINOPHIL RECRUITMENT TO THE AIRWAY
Patients recruited for studies involving SBP undergo a number of examinations before partici-
pation. All subjects must .demonstrate an immediate skin test response to antigen on skin testing. Furthermore, they must show a fall (-> 20%) in FEV 1 with antigen bronchoprovocation, even if they do not have clinical asthma. Because many of our studies are performed with patients with allergic rhinitis, a late allergic reaction is less predictable than in asthma. Despite responses that indicate an allergic airway reaction to a relevant antigen, it is not always possible to predicate which subject will manifest a late response (48 hours after antigen challenge) that includes significant eosinophilia. Evidence suggests that a late-phase response (i.e., a fall in FEV1 of 15% or more approximately 6 hours after challenge) would be a good predictor of airway eosinophilia. Thus we were interested in whether an in vitro test could identify those subjects who are more prone to have significant eosinophilia in response to antigen. In preliminary studies we wished to determine whether in vitro antigen-dependent basophil histamine release correlated with the eosinophil infiltration after SBP with allergen. That is, could we use basophil histamine release to antigen as a predictor of the patient who would manifest a significant eosinophilia to antigen? In this study allergic subjects without asthma were skin tested with common inhalant allergens. All had at least three positive prick skin test reactions. Peripheral blood samples were drawn, and the mononuclear cell band containing baosphils was isolated. Cells were then incubated with relevant antigen for 20 minutes and the
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s u p e r n a t a n t s c o l l e c t e d a n d a s s a y e d for h i s t a m i n e . T h e p a t i e n t s t h e n u n d e r w e n t a SBP a n d lavage with t h e s a m e a n t i g e n t h a t was u s e d for b a s o p h i l histamine release. Airway segments were identified by b r o n c h o s c o p y a n d c h a l l e n g e d with e i t h e r saline s o l u t i o n o r antigen, a n d lavage was p e r f o r m e d w i t h i n 5 minutes. T h e s a m e subjects w e r e s e e n again 48 h o u r s later; b r o n c h o s c o p y was rep e a t e d , a n d t h e s a m e s e g m e n t s w e r e lavaged. T h e s a m p l e s w e r e a n a l y z e d for h i s t a m i n e , a n d cell counts were made. I n t h e s e subjects t h e following o b s e r v a t i o n s w e r e m a d e . A l t h o u g h a high d e g r e e of b a s o p h i l h i s t a m i n e r e l e a s e ( > 2 0 % ) to a n t i g e n was n o t an a b s o l u t e m e a s u r e o f t h e 4 8 - h o u r airway r e s p o n s e to in vivo a n t i g e n challenges, low b a s o p h i l histam i n e r e l e a s e i n d i c a t e d (1) a low infiltration in t o t a l B A L cells, (2) a low p e r c e n t a g e o f B A L eosinophils, a n d (3) a low a b s o l u t e n u m b e r of B A L eosinophils. T h e factors d e t e r m i n i n g this c o r r e l a t i o n a r e n o t a p p a r e n t b u t raise a n u m b e r o f possibilities. First, t h e c e l l u l a r r e s p o n s e o f b a s o p h i l s , as m e a s u r e d by h i s t a m i n e release, m a y m i r r o r similar i n f l a m m a tory events in t h e airways, p a r t i c u l a r l y in t h e late allergic reactions. S e c o n d , b a s o p h i l h i s t a m i n e release m a y p a r a l l e l o t h e r i m m e d i a t e cell activation a n d i n d i c a t e an activation o f t h o s e p r o c e s s e s w h i c h also r e c r u i t e o s i n o p h i l s to t h e airways. B e c a u s e t h e r e was no c o r r e l a t i o n b e t w e e n i m m e d i a t e m a s t cell r e s p o n s e s a n d e o s i n o p h i l r e c r u i t m e n t , this corr e l a t i o n m a y b e l i m i t e d to t h e b a s o p h i l a n d reflected in b a s o p h i l h i s t a m i n e release. Finally, t h e s e m a y b e r a n d o m events. B e c a u s e t h e relative d e g r e e o f r e s p o n s e is similar in cells o f c o m m o n lineage, t h e possibility is r a i s e d t h a t t h e r e s p o n s e o f o n e cell (i.e., t h e b a s o p h i l ) to a n t i g e n m a y b e similar to t h a t which occurs in a n o t h e r celt, t h e e o s i n o p h i l . F u r t h e r m o r e , t h e d e f e c t a s s o c i a t e d with e n h a n c e d b a sophil r e l e a s a b i l i t y m a y b e a clue to a l t e r e d signal t r a n s d u c t i o n in cells in a t o p i c d i s e a s e in g e n e r a l . M o r e o v e r , t h e s e o b s e r v a t i o n s i n d i c a t e h o w little we k n o w a b o u t t h e b a s o p h i l a n d its r e l e v a n c e to allergic i n f l a m m a t i o n . REFERENCES
1. Kuna P, Reddigari SR, Schall TJ, Rucinski D, Sadick M, Kaplan AP. Characterization of the human basophil response to cytokines, growth factors, and histamine releasing factors of the intercrine/chemokine family. J Immunol 1993;150:1932-43. 2. Conroy MC, Adkinson NF Jr, Lichtenstein LM. Measurement of IgE on human basophils: relationship to serum IgE and anti-lgE-induced histamine release. J Immunol 1977;118:1317-21.
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3. Marone G, Poto S, Giugliano R, Celestino D, Bonic S. Control mechanisms of human basophil releasability. J ALLERGYCLINIMMLrNOL1986;78:974-80. 4. Butler JM, Chan SC, Stevens S, Hanifin JM. Increased leukocyte histamine released with elevated cyclic AMP: phosphodiesterase activity in atopic dermatitis. J ALLERGY CLIN IMMUNOL1983;71:490-7. 5. Butler JM, Ebertz M, Chan SC, Stevens SR, Sobieszczuk B, Hanifin JM. Basophil histamine release in atopic dermatitis and its relationship to disordered cyclic nucleotide metabolism. Acta Derm Venereol Suppl (Stockh) 1985; 144:55-60. 6. Ricketti AJ, Greenberger PA, Pruzansky JJ, Patterson R. Hyperreactivity of mediator-releasing cells from patients with allergic bronchopulmonary aspergillosis as evidence by basophil histamine release. J ALLERGYCLIN IMMUNOL 1983;72:386-92. 7. May CD. High spontaneous release of histamine in vitro from leukocytes of persons hypersensitive to foods. J ALLERGYCLIN IMMUNOL1976;58:432-7. 8. Skov PS, Norn S, Weeke B. Basophil histamine release in patients with bay fever: results compared with specific IgE and total IgE during immunotherapy. Clin Exp Immunol 1977;27;432-9. 9. Busse WW, Swenson CA, Sharpe G, Koschat M. Enhanced basophil histamine release to concanavalin A in allergic rhinitis. J ALLERGYCLIN IMMUNOL1986;78:90-7. 10. Kern F, Lichtenstein LM. Defective histamine release in chronic urticaria. J Clin Invest 1976;57:1369-77. 11. Findlay SR, Lichtenstein LM. Basophil "releasability" in patients with asthma. Am Rev Respir Dis 1980;122:53-9. 12. Tung R, Lichtenstein LM. In vitro histamine release from basophils of asthmatic and atopic individuals in D20. J Immunol 1982;128:2067-72. 13. Peters SP, Tung RS, Chatlam M, Bleeker ER, Lichtenstein L. Inhibitability and enbanceability of basophil histamine release in asthmatic and normal subjects. Int Arch Allergy Appl Immunol 1985;76:344-9. 14. Gaddy JN, Busse WW. Enhanced IgE-dependent basophil histamine release and airway reactivity in asthma. Am Rev Respir Dis 1986;134:969-74. 15. Casale TB, Wood D, Richerson HB, et al. Elevated bronchoalveolar lavage fluid histamine levels in allergic asthmatics are associated with methacholine bronchial hyperresponsiveness. J Clin Invest 1987;79:1197-203. 16. Jarjour NN, Calhoun WJ. Bronchoalveolar lavage in stable asthmatics does not cause pulmonary inflammation. Am Rev Respir Dis 1990;142:100-3. 17. Naclerio RM, Proud D, Togia AG, et al. Inflammatory mediators in late antigen-induced rhinitis. N Engl J Med 1985;313:65-70. 18. Bascom R, Wachs M, Naclerio RM, Pipkorn U, Galli SJ, Lichtenstein LM. Basophil influx occurs after nasal antigen challenge: effects of topical corticosteroid pretreatment. J ALLERGYCLIN IMMUNOL1988;81:580-9. 19. Liu MC, Hubbard WC, Proud D, et al. Immediate and late pulmonary responses to ragweed antigen challenge of the peripheral airways in allergic asthmatics. Am Rev Respir Dis 1991;144:51-8. 20. Sedgwick JB, Calhoun WJ, Gleich GJ, et al. Immediate and late airway response of allergic rhinitis patients to segmental antigen challenge. Am Rev Respir Dis 1991; 144:1274-81.
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26. Talbot SF, Atkins PC, Valenzano M, et al. Correlations of in vivo mediator release with late cutaneous allergic response in humans. I. Kinetics of histamine release. J ALLERGYCLIN IMMUNOL1984;74:819-26. 27. Shalit M, Schwartz LB, Golzar N, et al. Release of histamine and tryptase in vivo after prolonged cutaneous challenge with allergen in humans. J Immunol 1988;141: 821-6. 28. Talbot SF, Atkins PC, Goetzl EJ, et al. Accumulation of leukotriene C4 and histamine in human allergic skin reactions. J Clin Invest 1985;76:650-6. 29. Shalit M, Valone FH, Atkins PC, et al. Late appearance of phospholipid platelet-activating factor and leukotriene B4 in human skin after repeated antigen challenge. J ALLERCY CLIN IMMUNOL1989;83:691-6. 30. Tamura N, Agrawal DK, Suliaman, et al. Effects of platelet activating factor on the chemotaxis of normodense eosinophils from normal subjects. Biochem Biophys Res Commun 1987;142:638-44. 31. Chilton FH, Connell TR. 1-0-Alkyl-2-arachidonoyl-snglycero-3-phosphocholine: a common source of plateletactivating factor and arachidonate in human polymorphonuclear leukocytes. J Biol Chem 1984;259:12014-9. 32. Bochner BS, Charlesworth EN, Kagey-Sobotka A, et al. Interleukin-1 is released at sites of human cutaneous allergic reactions. J ALLERGYCLINIMMUNOL1990;86:830-9. 33. Schleimer RP, Rutledge BK. Cultured human vascular endothelial cells acquire adhesiveness for neutrophils after stimulation with interleukin 1, endotoxin, and tumorpromoting phorbol esters. J Immunol 1986;135:649-54. 34. Beviliaqua MP, Pober JS, Mendrick, et al. Identification of an inducible endothelial-leukocyte adhesion molecule. Proc Natl Acad Sci USA 1987;84:9238-42. 35. Subramanian N, Bray MA. Interleukin 1 releases histamine from human basophils and mast cells in vitro. J Immunol 1987;138:271-5. 36. Haak-Frendscho M, Dinarello CA, Kaplin AP, et al. Recombinant human interleukin 1 beta causes histamine release from human basophils. J ALLEROVCLIN IMMUNOL 1988;82:218-23. 37. Lee CE, Neuland ME, Teaford HG, et al. Interleukin-6 is released in the cutaneous response to allergen challenge in atopic individuals. J ALLERGYCLIN IMMUNOL1992;89: 1010-20.
38. Moller A, Lippert U, Lessmann D, et al. Human mast cells produce IL-8. J Immunol 1993;151:3261-6. 39. Bradding P, Feather IH, Wilson S, et al. Immunolocalization of cytokines in the nasal mucosa of normal and perennial rhinitic subjects: the mast cell as a source of IL-4, IL-5, and IL-6 in human allergic mucosal inflammation. J Immunol 1993;151:3853-65. 40. Morris FM, Whisman BA, Charlesworth EN. Measurement of TNF/alpha, GM-CSF, and IL-4 in the cutaneous late-phase response to antigen [Abstract]. J ALLERGYCLIN IMMUNOL1994;93:261. 41. Massey W, Friedman B, Kato M, et al. Appearance of granulocytes-macrophage colony stimulating factor activity at allergen-challenged cutaneous late-phase reaction sites. J Immunol 1993;150:1084-92. 42. Varney VA, Hamid QA, Gaga M, et al. Influence of grass pollen immunotherapy on cellular infiltration and cytokine mRNA expression during allergen-induced late-phase cutaneous responses. J Clin Invest 1993;92:644-51. 43. Pipkorn U, Proud D, Lichtenstein LM, et al. Effect of short-term glucocorticoid treatment on human nasal mediator release after antigen challenge. J Clin Invest 1987; 80:957-61. 44. Charlesworth EN, Kagey-Sobotka A, Schleimer RP, et al. Prednisone inhibits the appearance of inflammatory mediators and the influx of eosinophils and basophils associated with the cutaneous late-phase response to allergen. J Immunol 1991;146:671-6. 45. Schleimer RP, Lichtenstein LM, Gillespie E. Inhibition of basophil histamine release by anti-inflammatory steroids. Nature 1981;292:454-5. 46. Fadel R, Herpin-Richard N, Rihoux JP, Henolg E. Inhibitory effect of cetirizine 2 HC1 on eosinophilia in vivo. Clin Allergy 1987;17:373-6. 47. Charlesworth EN, Massey WA, Kagey-Sobotka A, Norman PS, Lichtenstein LM. Effect of H1 receptor blockade on the early and late response to cutaneous allergen challenge. J Pharmacol Exp Ther 1992;262:964-70. 48. Alam R, Dejarnatt A, Stafford S, et al. Selective inhibition of the cutaneous late but not immediate allergic response to antigens by misoprostol, a PGE analog. Am Rev Respir Dis 1993;148:1066-70.
Eosinophils and basophils in allergic airway inflammation William W. Busse, MD, Julie B. Sedgwick, Nizar N. Jarjour, and William J. Calhoun Madison, Wis.
From the Section of Allergy and Clinical Immunology, Department of Medicine, University of Wisconsin. J ALLERGYCUN IMMVNOL1994;94:1250-4. Copyright © 1994 by Mosby-Year Book, Inc. 0091-6749/94/$3.00 + 0 1/0/59927
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A i r w a y i n f l a m m a t i o n h a s b e e n a c c e p t e d as a feature of asthma and other allergic inflammatory d i s e a s e s . T h e m a n y cells p a r t i c i p a t i n g in this p r o c e s s i n c l u d e m a s t cells, b a s o p h i l s , e o s i n o p h i l s , a n d l y m p h o c y t e s . O f t h e s e cells t h e m a s t cell, e o s i n o -
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Abbreviations used
BAL: Bronchoalveolar lavage FEVI: Forced expiratory volume in 1 second PC2o: Provocative concentration needed to decrease FEV 1 by 20% PGD2: Prostaglandin D2 SBP: Segmental bronchoprovocation
±
=x 0 24 E o~ 18 e-
phil, and lymphocyte have attracted greatest attention. Further, these cells are easiest to study in the in vivo setting because they possess measurable mediators, characteristic morphology, and distinct surface receptors, and because they can be isolated in sufficient number for in vitro study. In contrast, the basophil is a less dominant cell both in circulation and airway secretions. Thus efforts to define its unique or interactive contribution to allergic inflammation have been more difficult and insights obtained remain less specific. The following article discusses evidence that basophil function may have a correlation with markers of asthma severity, participate in the late allergic airway reaction, and predict eosinophilic participation in the airway's response to allergen. BASOPHIL HISTAMINE RELEASE AND AIRWAY RESPONSIVENESS Many mechanisms determine basophil histamine release, including membrane-bound IgE and cytokine exposure. 1 However, the absolute concentration of membrane-bound IgE does not correlate with the degree of basophil histamine released after an IgE-dependent stimulus2; consequently it has been proposed that basophil secretion may be determined by an intrinsic cellular property defined as "releasability. ''3 A number of features of basophil releasability have already been determined. Furthermore, abnormalities in basophil release of histamine have been noted in atopic dermatitis, 4' 5 allergic bronchopulmonary aspergillus, 6 food hypersensitivity, 7 allergic rhinitis, 8" 9 chronic urticaria, 1° and asthma. 1144 Although these associations show a potential relationship to allergic diseases, they do not identify the mechanism of enhanced releasability. As will be discussed, basophil histamine release may participate in the process airway inflammation and be linked to or contribute to airway responsiveness. In previous work we expanded on earlier observations and evaluated basophil histamine release in subjects with asthma. 14 The premise of
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HIGH
BAL SEGMENT (AG DOSE) FIG. 1. Comparison of leukocyte histamine release to anti-lgE (10 to 1000 ng/ml) in six normal subjects, five nonallergic patients with asthma and seven allergic patients with asthma. Values represent mean (-+SEM) percentage histamine release. Asterisks denote degree of significance when asthma groups are compared with normal group. Spontaneous histamine release was subtracted from simulated release values. (From Gaddy JN, Busse WW. A m Rev Respir Dis 1986;134:969-74.)
this study was to determine whether any relationship existed between IgE-dependent basophil histamine releasability and a pathognomonic feature of asthma, bronchial responsiveness. In this study 16 patients with asthma were selected for examination. Of these, nine demonstrated immediate skin test reactivity to common inhalant allergens; the other seven had no evidence of skin test reactivity and were considered to have nonallergic asthma. In addition, seven control subjects were identified who were age matched but who had no evidence of allergic disease or asthma. With the use of anti-IgE as the basophil-secretagogue, both allergic and nonallergic asthmatic subjects released histamine, as did the nonallergic control group (Fig. 1). At all concentrations of anti-IgE used, basophils from both asthma populations, allergic and nonallergic, released more histamine than did the control population. These data indicate that basophil histamine release may not be dictated alone by the concentration of surface-bound IgE. Rather, other factors (i.e., basophil releasability) were likely to be important. Furthermore, enhanced basophil releasability was a feature of asthma and not determined by the presence of allergic disease. To extend these observations we evaluated the relationship of basophil histamine release to a
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FIG. 2. Correlation between anti-lgE-dependent leukocyte histamine release and PC2ofor aerosol histamine was evaluated in 12 subjects with asthma (eight allergic, four nonallergic). Values represent percentage of histamine release at anti-lgE concentration of 30 ng/ml versus log e PC2o for each subject. There was significant (p < 0.05) inverse correlation as determined by linear regression analysis and Pearson's rank correlation between percentage of histamine release and log e PC2o. (From Gaddy JN, Busse WW. Am Rev Respir Dis 134:969-74.)
marker of asthma: airway responsiveness to histamine measured by the provocative concentration required to decrease the forced expiratory volume in 1 second (FEV0 by 20% (PC20). A significant inverse correlation was noted between PC20 to histamine and three concentrations of anti-IgE: 30.0 ng/ml (r = -0.753), 100 ng/ml (r = -0.678), and 300 ng/ml (r = -0.671) (Fig. 2). These data indicate that the greater the level of basophil histamine release to anti-IgE, the more responsive to inhaled histamine was the patient with asthma. Although these data do not indicate that basophil histamine releasability causes bronchial hyperresponsiveness, it raised the question whether this feature of the basophil may be a reflection of similar abnormalities in the airway of patients with asthma. It is intriguing to consider the possibility that basophil releasability and airway hyperresponsiveness share a same intrinsic abnormality in signal transduction, and this abnormality may account for releasability and responsiveness. EVIDENCE THAT BASOPHILS PARTICIPATE IN THE LATE AIRWAY ALLERGIC REACTION
Mast cells and baosphils are cellular sources of airway histamine. The mast cell makes the dominant contribution to the airway histamine "pool." Casale et al. 15 and other investigators 16 found increased bronchoalveolar lavage (BAL) concentrations of histamine in patients with allergic
asthma when compared with those with allergic rhinitis and with normal subjects. Although both patients with allergic asthma and those with rhinitis had increased concentrations of mast cells in the lavage fluid compared with normal subjects, there was no correlation between BAL mast cell members and lavage fluid concentration of histamine. No mention was made of the number of basophils in the airway lavage fluid. In addition, BAL histamine concentrations were greater in those subjects with increased airway responsiveness. These data raise the possibility that ongoing mediator release and presumably other factors of allergic inflammation contribute to altered airway function in asthma, and the airway fluid concentration of histamine is a marker of this process. The relative contribution of mast cells and basophils to this process was not clarified. Evidence for basophil participation in the allergic inflammatory process first came from work by Naclerio et al. 17 Antigen challenge of the nose elicits an immediate response of sneezing and mediator release, including histamine, kinins, tosyl-arginine methyl esterases, and prostaglandin Dz (PGD2). In contrast, the late response to antigen is characterized by an increase in histamine but not in PGD2. Because PGD2 is derived from mast cells alone, the late-phase rise in histamine in nasal fluids was attributed to basophil recruitment to the nose and activation. This conclusion has been substantiated by cell staining. 18 Similar findings have been noted in the lower part of the airway. Liu et a l . 19 conducted localized antigen challenge of airway segments by bronchoscopy. Lavage fluid obtained 5 minutes after antigen challenge had increased levels of histamine, PGD2, and kinins. A similar profile of mediator release increase was noted 19 hours after the antigen challenge; these data do not indicate which mediator-releasing cells contributed to the "late" responses to antigen. However, Liu et al. were able to show an increase in alcian bluestaining cells in the late-phase lavage fluid. Furthermore, the predominant alcian blue-staining cell in this latter category was the basophil. These data support the concept that basophils are recruited to the airway after antigen challenge and release mediators in the late phase of the allergic reaction. We have also used segmental bronchoprovocation (SBP) as a method to more clearly define the airway allergic response to antigen. 2° In our protocol isolated airways are challenged with saline
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-io
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I0
30 [Anti-IgE]
IO0
~500
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(ng/ml)
FIG. 3. Mediator release after antigen challenge. BAL fluids were assayed immediately (open bars, 12 minutes) and late (hatched bars, 48 hours) for histamine (top) and tryptase (bottom).
solution or relevant antigen. Lavage is performed immediately after antigen challenge (N 5 minutes) and again 48 hours later. There was a marked increase in histamine and tryptase immediately after antigen challenge (Fig. 3). The increase in these mast cell mediators was proportional to the concentration of antigen delivered to the airway. In contrast, when BAL fluids were obtained 48 hours after antigen challenge, tryptase values were similar to baseline concentrations of this mediator. Histamine concentrations in lavage fluid, however, showed a different pattern. Although the concentrations of histamine obtained immediately after antigen challenge were markedly elevated, lavage fluid histamine was still detectable at 48 hours after antigen challenge and concentrations were significantly elevated over baseline values. Because tryptase was not detected in the 48-hour postchallenge lavage sample, we concluded that a probable cell source of histamine was the basophil. This conclusion has to remain presumptive because we did not detect basophils in cells from the late lavage fluid. Nonetheless, in light of data from Johns Hopkins investigations that basophils are a component of the late response to antigen, we speculate that these late-phase increases in histamine reflect a basophil contribution to the airway allergic response. PERIPHERAL BLOOD BASOPHIL HISTAMINE RELEASE AND EOSINOPHIL RECRUITMENT TO THE AIRWAY
Patients recruited for studies involving SBP undergo a number of examinations before partici-
pation. All subjects must .demonstrate an immediate skin test response to antigen on skin testing. Furthermore, they must show a fall (-> 20%) in FEV 1 with antigen bronchoprovocation, even if they do not have clinical asthma. Because many of our studies are performed with patients with allergic rhinitis, a late allergic reaction is less predictable than in asthma. Despite responses that indicate an allergic airway reaction to a relevant antigen, it is not always possible to predicate which subject will manifest a late response (48 hours after antigen challenge) that includes significant eosinophilia. Evidence suggests that a late-phase response (i.e., a fall in FEV1 of 15% or more approximately 6 hours after challenge) would be a good predictor of airway eosinophilia. Thus we were interested in whether an in vitro test could identify those subjects who are more prone to have significant eosinophilia in response to antigen. In preliminary studies we wished to determine whether in vitro antigen-dependent basophil histamine release correlated with the eosinophil infiltration after SBP with allergen. That is, could we use basophil histamine release to antigen as a predictor of the patient who would manifest a significant eosinophilia to antigen? In this study allergic subjects without asthma were skin tested with common inhalant allergens. All had at least three positive prick skin test reactions. Peripheral blood samples were drawn, and the mononuclear cell band containing baosphils was isolated. Cells were then incubated with relevant antigen for 20 minutes and the
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s u p e r n a t a n t s c o l l e c t e d a n d a s s a y e d for h i s t a m i n e . T h e p a t i e n t s t h e n u n d e r w e n t a SBP a n d lavage with t h e s a m e a n t i g e n t h a t was u s e d for b a s o p h i l histamine release. Airway segments were identified by b r o n c h o s c o p y a n d c h a l l e n g e d with e i t h e r saline s o l u t i o n o r antigen, a n d lavage was p e r f o r m e d w i t h i n 5 minutes. T h e s a m e subjects w e r e s e e n again 48 h o u r s later; b r o n c h o s c o p y was rep e a t e d , a n d t h e s a m e s e g m e n t s w e r e lavaged. T h e s a m p l e s w e r e a n a l y z e d for h i s t a m i n e , a n d cell counts were made. I n t h e s e subjects t h e following o b s e r v a t i o n s w e r e m a d e . A l t h o u g h a high d e g r e e of b a s o p h i l h i s t a m i n e r e l e a s e ( > 2 0 % ) to a n t i g e n was n o t an a b s o l u t e m e a s u r e o f t h e 4 8 - h o u r airway r e s p o n s e to in vivo a n t i g e n challenges, low b a s o p h i l histam i n e r e l e a s e i n d i c a t e d (1) a low infiltration in t o t a l B A L cells, (2) a low p e r c e n t a g e o f B A L eosinophils, a n d (3) a low a b s o l u t e n u m b e r of B A L eosinophils. T h e factors d e t e r m i n i n g this c o r r e l a t i o n a r e n o t a p p a r e n t b u t raise a n u m b e r o f possibilities. First, t h e c e l l u l a r r e s p o n s e o f b a s o p h i l s , as m e a s u r e d by h i s t a m i n e release, m a y m i r r o r similar i n f l a m m a tory events in t h e airways, p a r t i c u l a r l y in t h e late allergic reactions. S e c o n d , b a s o p h i l h i s t a m i n e release m a y p a r a l l e l o t h e r i m m e d i a t e cell activation a n d i n d i c a t e an activation o f t h o s e p r o c e s s e s w h i c h also r e c r u i t e o s i n o p h i l s to t h e airways. B e c a u s e t h e r e was no c o r r e l a t i o n b e t w e e n i m m e d i a t e m a s t cell r e s p o n s e s a n d e o s i n o p h i l r e c r u i t m e n t , this corr e l a t i o n m a y b e l i m i t e d to t h e b a s o p h i l a n d reflected in b a s o p h i l h i s t a m i n e release. Finally, t h e s e m a y b e r a n d o m events. B e c a u s e t h e relative d e g r e e o f r e s p o n s e is similar in cells o f c o m m o n lineage, t h e possibility is r a i s e d t h a t t h e r e s p o n s e o f o n e cell (i.e., t h e b a s o p h i l ) to a n t i g e n m a y b e similar to t h a t which occurs in a n o t h e r celt, t h e e o s i n o p h i l . F u r t h e r m o r e , t h e d e f e c t a s s o c i a t e d with e n h a n c e d b a sophil r e l e a s a b i l i t y m a y b e a clue to a l t e r e d signal t r a n s d u c t i o n in cells in a t o p i c d i s e a s e in g e n e r a l . M o r e o v e r , t h e s e o b s e r v a t i o n s i n d i c a t e h o w little we k n o w a b o u t t h e b a s o p h i l a n d its r e l e v a n c e to allergic i n f l a m m a t i o n . REFERENCES
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J ALLERGY CLIN IMMUNOL DECEMBER 1994
3. Marone G, Poto S, Giugliano R, Celestino D, Bonic S. Control mechanisms of human basophil releasability. J ALLERGYCLINIMMLrNOL1986;78:974-80. 4. Butler JM, Chan SC, Stevens S, Hanifin JM. Increased leukocyte histamine released with elevated cyclic AMP: phosphodiesterase activity in atopic dermatitis. J ALLERGY CLIN IMMUNOL1983;71:490-7. 5. Butler JM, Ebertz M, Chan SC, Stevens SR, Sobieszczuk B, Hanifin JM. Basophil histamine release in atopic dermatitis and its relationship to disordered cyclic nucleotide metabolism. Acta Derm Venereol Suppl (Stockh) 1985; 144:55-60. 6. Ricketti AJ, Greenberger PA, Pruzansky JJ, Patterson R. Hyperreactivity of mediator-releasing cells from patients with allergic bronchopulmonary aspergillosis as evidence by basophil histamine release. J ALLERGYCLIN IMMUNOL 1983;72:386-92. 7. May CD. High spontaneous release of histamine in vitro from leukocytes of persons hypersensitive to foods. J ALLERGYCLIN IMMUNOL1976;58:432-7. 8. Skov PS, Norn S, Weeke B. Basophil histamine release in patients with bay fever: results compared with specific IgE and total IgE during immunotherapy. Clin Exp Immunol 1977;27;432-9. 9. Busse WW, Swenson CA, Sharpe G, Koschat M. Enhanced basophil histamine release to concanavalin A in allergic rhinitis. J ALLERGYCLIN IMMUNOL1986;78:90-7. 10. Kern F, Lichtenstein LM. Defective histamine release in chronic urticaria. J Clin Invest 1976;57:1369-77. 11. Findlay SR, Lichtenstein LM. Basophil "releasability" in patients with asthma. Am Rev Respir Dis 1980;122:53-9. 12. Tung R, Lichtenstein LM. In vitro histamine release from basophils of asthmatic and atopic individuals in D20. J Immunol 1982;128:2067-72. 13. Peters SP, Tung RS, Chatlam M, Bleeker ER, Lichtenstein L. Inhibitability and enbanceability of basophil histamine release in asthmatic and normal subjects. Int Arch Allergy Appl Immunol 1985;76:344-9. 14. Gaddy JN, Busse WW. Enhanced IgE-dependent basophil histamine release and airway reactivity in asthma. Am Rev Respir Dis 1986;134:969-74. 15. Casale TB, Wood D, Richerson HB, et al. Elevated bronchoalveolar lavage fluid histamine levels in allergic asthmatics are associated with methacholine bronchial hyperresponsiveness. J Clin Invest 1987;79:1197-203. 16. Jarjour NN, Calhoun WJ. Bronchoalveolar lavage in stable asthmatics does not cause pulmonary inflammation. Am Rev Respir Dis 1990;142:100-3. 17. Naclerio RM, Proud D, Togia AG, et al. Inflammatory mediators in late antigen-induced rhinitis. N Engl J Med 1985;313:65-70. 18. Bascom R, Wachs M, Naclerio RM, Pipkorn U, Galli SJ, Lichtenstein LM. Basophil influx occurs after nasal antigen challenge: effects of topical corticosteroid pretreatment. J ALLERGYCLIN IMMUNOL1988;81:580-9. 19. Liu MC, Hubbard WC, Proud D, et al. Immediate and late pulmonary responses to ragweed antigen challenge of the peripheral airways in allergic asthmatics. Am Rev Respir Dis 1991;144:51-8. 20. Sedgwick JB, Calhoun WJ, Gleich GJ, et al. Immediate and late airway response of allergic rhinitis patients to segmental antigen challenge. Am Rev Respir Dis 1991; 144:1274-81.