Original Article. correlation between bronchial hyperresponsiveness and in vitro cytokine production was noted in asthma ABSTRACT

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1 Allergology International (1998) 47: Original Artile Bronhial and nasal responsiveness in atopi asthma and allergi rhinitis patients: Relationship of loal responsiveness to ytokine prodution by peripheral blood mononulear s Kei]i Maeda, l Toshio Tonoko,? Yoshinobu Katada,2 Arata Horii," Kimihiro Nose," Hiroshi Ohi.? Satoshi Oqino," Masaki Suernuro,? Tadamitsu Kishimoto? and Tsuyoshi lqoroshi' 'Seond Department of Internal Mediine and 3Department of Otolaryngology, Osaka Teishin Hospital and Departments of 21nternal Mediine III and "Otolrvnqoloqv, Osaka University Medial Shool, Osaka, Japan ABSTRACT To investigate the relationship between loal responsiveness and allergi symptoms, bronhial and nasal responsiveness were measured in the following four groups of subjets: (i) bronhial asthma patients with serum house dust mite (HOM)-speifi Ig antibody; (ii) allergi rhinitis patients with serum HOM-speifi Ig antibody; (iii) normal ontrol subjets with HOM-speifi Ig antibody; and (iv) normal ontrol subjets without Ig antibody speifi for 1 ommon aero-allergens. Bronhial hyperresponsiveness was deteted in all subjets with asthma (group 1) and in some subjets from groups 2 and 3, but not in subjets from group 4. Nasal hyperresponsiveness was found in all subjets with allergi rhinitis (group 2) and in some subjets from groups 1 and 3, but not in subjets from group 4. These findings indiate that loal hyperresponsiveness of the non-diseased organ is influened by an individual's atopi status. Interleukin (IL)-4 and IL-5 prodution by peripheral blood mononulear s (PBMC) was measured after stimulation with HOM in groups 1, 2 and 3 and was found to be similar in all three groups. A Correspondene: Dr Keiji Maeda, Seond Department of Internal Mediine, Osaka Teishin Hospital, Karasugatsuji, Tennohji-ku, Osaka 543, Japan. Reeived 5 Marh Aepted for publiation 11 September orrelation between bronhial hyperresponsiveness and in vitro ytokine prodution was noted in asthma patients. These results suggest that the apaity of IL-4 or IL-5 prodution by PBMC may reflet loal hyperresponsiveness in ase of asthma. Key words: allergi rhinitis, atopy, bronhial asthma, bronhial responsiveness, interleukin-4, interleukin-5, nasal responsiveness INTRODUCTION Bronhial asthma (BA) is a disease haraterized by inreased bronhial reativity. 1 Patients with perennial rhinitis have a hyperreative nasal muosa (nasal hyperreativity).2,3 It has been suggested that allergi rhinitis patients who have bronhial hyperresponsiveness are more likely to develop asthma. 4,s In ontrast, it has not been learly determined whether BA patients have nasal hyperreativity, despite the strong epidemiologi assoiations between atopi asthma and allergi rhinitis." Although inreased airway responsiveness has been reported in patients with allergi rhinitis who show no asthma syrnptorns.v' it is not lear whether the assoiation is predominantly with rhinitis itself or with the atopi status of the rhinitis patients. Of all potential allergens in the environment assoiated with asthma, allergy to house dust mite (HOM) is the

2 46 K MADA T AL. most ommon independent risk fator for the development of the disese." It is also known that HOM is a signifiant perennial indoor allergen that an preipitate allergi rhinitis." In the present study we used serum Ig antibody titer to HOM to evaluate atopi status. To investigate the assoiation between loal responsiveness and allergi symptoms among individuals with serum HOM-speifi Ig antibody, we studied four groups of patients and normal subjets. Our purpose was to determine how loal responsiveness (bronhial or nasal) and ytokine prodution from peripheral blood mononulear els (PBMC) influened the ourrene of BA and allergi rhinitis. MTHODS Subjets Subjets with the following harateristis were hosen randomly from our linis (Internal Mediine and Otolaryngology, Osaka Teishin Hospital and Osaka University Medial Shool, Osaka, Japan) and volunteers: (i) patients with a linial diagnosis of BA who had no history of allergi rhinitis and had elevated serum levels of HOM-speifi Ig antibody (n = 13); (ii) patients with a linial diagnosis of allergi rhinitis who had no history of asthma and had elevated serum levels of HOM-speifi Ig antibody (n = 11); (iii) volunteers working at our hospital with no history of asthma or allergi rhinitis who had elevated serum levels of HOM-speifi Ig antibody (n = 1); and (iv) volunteers working at our hospital with no speifi Ig antibody for the following 1 aero-allergens (n = 9): Dermatophagoides pteronyssinus (Der p), Dermatophagoides farinae (Der ~, house dust 1, house dust 2, ragweed pollen, orhard grass, Aspergillus fumigatus, Candida albians, Alternaria alternata and Japanese edar pollen. The linial harateristis of the subjets are shown in Table 1. Speifi Ig antibodies were measured with a Pharmaia CAP System (Pharmaia, Uppsala, Sweden) by external ommerial laboratories. A CAP-RAST lass of 2 or more was onsidered positive (lass, <.34 UA/mL; lass 1, UA/mL; lass 2, UA/mL; lass 3, UA/mL; lass 4, UA/mL; lass 5, UA/mL; lass 6, > 1 UA/mL). House dust mite was Der f in both the CAP-RAST assay and in the in vitro studies. Fored vital apaity and fored expiratory volume in 1 s were measured using a spirometer (Autospiro AS-5; MINATO Ltd, Osaka, Japan). Bronhial and nasal responsiveness Bronhial responsiveness was measured with an Astograph (Chest Corp., Tokyo, Japan, a diret-writing reorder for respiratory resistane (Rrs) dose-response urves, during ontinuous inhalation of methaholine at stepwise inremental onentrotions.!" Respiratory resistane was measured by the fored osillation method. It showed a urvilinear inrease with an inrease Table 1. Clinial harateristis of subjets --- Group 1 Group 2 Group 3 Group 4 ~_._._-~_.~_._ _._--~---_._~_.~ ~--- No. subjets Gender (M/F) 6/7 3/8 6/4 5/4 Smoker/non-smoker 2/11 2/9 2/8 3/6 Age (years)* 44.7 (17.9) 34.9 (14.6) 37. (15.5) 34.9 (13.5) CAP-RAST sore for 3.31 (1.11) 3.54 (.93) 3.5 (1.8) HOM (Der~* Serum Ig level 399 (26-12) 38 (24-71) 889 (32-44) 68 (7-2) (IU/mL)** osinophil ount in 513 (2-15) 314 (-64) 295 (65-67) 121 (61-24) peripheral blood (lmm 3)** % FVC* 89.6 (14.) 95.4 (13.) 95.6 (7.2) 91.8 (16.1) FV1o%* 81.5(13.1) 92. (6.3) 91.8 (7.7) 92.5 (5.3) *Oata are the mean with SO given in parentheses; **data are the mean with the range given in parentheses. % FVC, % of predited fored vital apaity; FV 1 %, % of predited fored expiratory volume in 1 s; HOM, house dust mite. Group 1, bronhial asthma patients with serum HOM-speifi Ig antibody; group 2, allergi rhinitis patients with serum HOM-speifi Ig antibody; group 3, ontrol subjets with no symptoms of allergi diseases who have serum HOM-speifi Ig antibody; group 4, ontrol subjets with no symptoms of allergi diseases and no speifi serum Ig antibody for a panel of 1 aero-allergens, inluding HOM.

3 LOCAL RSPONSIVNSS AND CYTOKINS 47 in the methaholine onentration. The minimum dose of methaholine (D min ) was used as an indiator of bronhial sensitivity and was defined as the umulative dose at the infletion point where the reiproal of Rrs (Grs) dereased linearly; D min was expressed in units equal to a 1. mg/ml aerosol inhalation of methaholine for 1 min. The existene of bronhial hyperresponsiveness was determined by a Dmin value < 5. 1 Drugs were disontinued at least 12 h before the study. Nasal responsiveness was measured by plaing a piee of round paper (Toyo-Roshi No.5, Tokyo, Japan) ontaining inreasing doses of histamine hloride over the surfae of the inferior turbinate bilterllv.' The logarithm of the histamine hloride onentration that indued sneezing and nasal seretion was defined as nasal responsiveness. Histamine hloride was prepared at onentrations ranging from 15 to 1 uq/rnl, by serial la-fold dilution. Bronhial responsiveness and nasal responsiveness were measured on separate days (1-2 weeks apart). All subjets were free from wheezing, ough, nasal disharge, sneezing and respiratory trat infetion for at least 3 weeks before the test. Fully informed onsent was obtained from all subjets partiipating before entry into the study. Reagents and s RPMI 164 (Gibo, Grand Island, NY, USA), supplemented with 1% fetal bovine serum (FBS; Gibo), 2 meraptoethanol (.5 mmolll), peniillin (1 U/mL) and streptomyin (.1 mg/ml), was used as the ulture medium. House dust mite (Der ~ extrat was provided by Torii Co. Ltd (Tokyo, Japan). Measurement of interleukin-4 and -5 prodution Peripheral blood mononulear s were isolated from heparinized venous blood by density gradient entrifugation on Fioll-Hypaque (Pharmaia, Uppsala, Sweden). Isolated mononulear s (2 X 1 6 /ml) were added to eah w of a 24-w ulture plate in.5 ml ulture medium with mite extrat (1 uq/ml}, whih was the optimal onentration for interleukin (IL)-4 svnthesis.!' After 48 h, the supernatant was olleted and the amount of IL-4 and IL-5 was measured by LISA (Compat Interleukin 4 LISA kit; CLB, Amsterdam, The Netherlands; LISA for human Interleukin 5; Immunoteh International, Marseilles, Frane, respetively). Using these assays,.1-1 pg/ml IL-4 or 2-1 pg/ml IL 5 was measured in the supernatants and was ompared with different onentrations of reombinant IL-4 or IL-5. Interleukin-4 and -5 prodution was measured within 6 weeks after the assessment of bronhial and nasal responsiveness. Statistial analysis The Student's t-test and the Mann-Whitney U-test were used for omparisons between groups. Correlations were investigated using Spearman's rank orrelation test. RSULTS Bronhial responsiveness All 13 asthma patients (group 1) showed bronhial -.. hyperresponsiveness (Dmin < 5; Fig. 1). Three of '" : :::l : '" Q.l Q.l.~ 1 -: '".. '" ~ o.s: ue o.1 - Group 1 Asthma HDM (+1 Group 2 Rhinitis HDM (+1 Group 3 Control HDM (+1 Group 4 Control RAST (-I Fig. 1 Individual values for the minimum dose of methaholine (Omin) in the four groups, where Omin is an indiator of bronhial responsiveness and Omin > 5 is normal.

4 48 K MADA T AL. rhinitis patients (group 2) and three of 1 HOM-speifi Ig positive normal ontrol subjets (group 3) also showed bronhial hyperresponsiveness. Overall, 55.9% of subjets with elevated levels of HOM-speifi Ig showed bronhial hyperresponsiveness. In ontrast, none of the nine normal ontrol subjets without speifi Ig (group 4) showed bronhial hyperresponsiveness. Nasal responsiveness In a preliminary study, normal individuals without a history of rhinitis showed a nasal responsiveness of 4 or 5. Most normal ontrol subjets without speifi Ig (7/9; group 4) also showed a nasal responsiveness of 4 or 5 (Fig. 2). Seven of 13 BA patients (group 1) and eight of 1 HOMspeifi Ig positive normal ontrol subjets (group 3) showed nasal hyperresponsiveness (sore ~ 3). All 11 rhinitis patients (group 2) showed nasal hyperresponsiveness and it was more prominent than in the other three groups. Overall, 76.4% of subjets with elevated levels of HOM-speifi Ig showed nasal hyperresponsiveness. Interleukin-4 and -5 prodution from PBMC Peripheral blood mononulear s from subjets in groups 1-3 were stimulated with HOM extrat in vitro and the IL-4 and -5 levels in ulture supernatants were <, ) CL U => \J 2 4 CL ""<:/', ~ !......L. - Group 1 Group 2 Group 3 Asthma Rh initis Control HDM l+l Q) Fig. 3 Interleukin (IL)-4 prodution by peripheral blood s stimulated with house dust mite. Mean values, expressed as the geometri mean, are represented by horizontal bars. - 2 Q).:: - Q. <, - ) ~ A.: C 2 - Z - - u 1 => \J Ci l() 1 - ~ Group 1 Group 2 Group 3 Group 4-4\ Group 1 Group 2 Group 3 Asthma Rhinitis Control Asthma Rhinitis Control Control HDM l+l HDM l+l HDM l+l HDM l+l RAST H Fig. 4 Interleukin (IL)-5 prodution by peripheral blood Fig. 2 Individual nasal responsiveness to histamine hloride mononulear s stimulated with house dust mite. Geometri in the four groups. A nasal responsiveness of 4 or 5 is normal. mean values are represented by horizontal bars.

5 LOCAL RSPONSIVNSS AND CYTOKINS 49 <, m Q. o U :::l - o.1 Q. """".J (a) 1.1-t-"""T""'"T"T'TTTnr-...,...,.,rrnrrrr---,...,.-T"T'I"T"Tfr"""""r-T'""""-'om (b) 1 <, m Q.. 1 o U :::l - o 1 Q. I.().J r~~--r..,...,...'f""T"T"M'lT"'"--...,..TTTTTll.1.1 Bronhial responsiveness (D min ; units) Fig. 5 (a) Relationship between interleukin (IL)-4 and the minimum dose of methaholine (Omin) in bronhial asthma (group 1). There is a signifiant negative orrelation (r = -.79; P <.1). (b) Relationship between IL-5 and Omin in bronhial asthma patients (group 1). There is a weak negative orrelation (r =.51; P =.7). measured (Figs 3,4). The mean IL-4 levels were 1.5 ± 1.8,1.8 ± 1.9 and 1.7 ± 2.1 pg/ml in groups 1,2 and 3, respetively, while the respetive IL-5 levels were found to be 64 ± 65, 16 ± 54 and 75 ± 81 pg/ml. There was no signifiant differene in the prodution of either ytokine among the different groups with HOM-speifi Ig positivity. Loal responsiveness and ytokine prodution To test whether loal responsiveness (bronhial and nasal responsiveness) may be influened by ytokine prodution in peripheral blood, the relationship between loal responsiveness and IL-4 or -5 prodution was investigated. There was no orrelation between nasal responsiveness and IL-4 or -5 prodution among the 1 subjets in eah of the groups 1-3 or among all subjets in these three groups ombined. However, the relationship between Omin (bronhial responsiveness) and IL-4 prodution in asthma patients (group 1) showed a strong negative orrelation (Fig. 5; r = -.79; P <.1), although no suh orrelation was found among all subjets who had bronhial hyperresponsiveness (Omin < 5) in groups 1-3. The relationship between Omin and IL-5 prodution in asthma patients showed a weaker negative orrelation (Fig. 5b; r = -.51; P =.7), while IL-5 and bronhial responsiveness showed no relationship among all subjets with bronhial hyperresponsiveness (Omin < 5) in groups 1-3. The relationship between Omin and ytokine prodution ould not be assessed separately in groups 2 or 3 beause only three subjets showed bronhial hyperresponsiveness in eah group. There was a strong positive orrelation between IL-4 and -5 prodution in BA patients (r =.66; P <.3; data not shown). DISCUSSION The results of the present study suggest an assoiation between atopy and subjets with an elevated loal hyperresponsiveness. Of the serum level of HOM-speifi Ig, all asthma patients (group 1) had bronhial hyperresponsiveness (Omin < 5). In addition, approximately 3% of both allergi rhinitis patients (group 2) and subjets with no allergi symptoms (group 3) had bronhial hyperresponsiveness. In ontrast, the normal ontrol subjets with no aero-allergen-speifi Ig (group 4) did not have bronhial hyperresponsiveness. Similar results were obtained in the ase of nasal responsiveness. Of the subjets with elevated serum HOM-speifi Ig, all rhinitis patients (group 2) had nasal hyperresponsiveness and more than half of both groups 1 and 3 also had moderate nasal hyperresponsiveness. Most normal ontrol subjets with no aero-allergen-speifi Ig had normal nasal responsiveness. Witt et /. have reported that the prevalene of bronhial histamime responsiveness in adults inreased from 5.8% in subjets who did not respond to allergen prik tests to 22.2% in subjets who responded to all five allergen qroups.'? Woolok et 1. reported that 7.3% of non-atopi subjets showed bronhial hyperresponsiveness ompared with 16.5% of those with severe atopy, and stated that there was a signifiant assoiation between bronhial responsiveness and atopy in random adult populotions.':' Our results are ompatible with these

6 5 K MADA TAL. findings and extend the assoiation with atopy to nasal hyperresponsiveness. It is w known that allergi rhinitis patients often have bronhial hvperresponsiveness;"? but whether BA patients have nasal hyperresponsiveness has been unlear. In aordane with our linial experiene, more than half the asthma patients with serum HOM-speifi Ig (group 1) showed nasal hyperresponsiveness in the present study. This may explain why asthma patients often omplain of nasal symptoms. Although bronhial responsiveness showed almost no overlap between asthma patients and non-atopi normal ontrol subjets, nasal responsiveness did not ompletely separate rhinitis patients from non-atopi normal ontrol subjets. This may be beause nasal responsiveness an be aquired readily after nasal infetion and rhinitis is more ommon than asthma. In any event, loal responsiveness of the non-diseased organ was inreased to some extent among serum HOM-speifi Ig positive subjets (groups 2 and 3 for bronhial responsiveness and groups 1 and 3 for nasal responsiveness), suggesting that loal responsiveness of the non-diseased organ may be affeted by an individual's atopi status. and Interleukin-4 is known to be a mast growth foetor" it also promotes B isotype swithing to Ig.15 Allergen-indued Ig-dependent mast degranulation may be responsible for aute bronhoonstritor responses in atopi sthrn.l? Interleukin-5 promotes the differentiation,17 vasular adhesion 18 and in vitro survival of eosinophils'? and eosinophil infiltration and ativation is known to ontribute to hroni bronhial hyperreotivitv." It has also been shown that atopi asthma is assoiated with inreased expression of mrna for IL-3, IL-4, IL-5 and granuloyte-marophage olony stimulating fator in the bronhi." It has also been reported that s expressing mrna for IL-4 and -5 inrease during the allergen-indued nasal response in allergi rhinitis. 22 Although inreases in IL-4 and IL-5 ativity has been shown in diseased organs (bronhial muosa or reovered s from bronhoalveolar lavage for BA and nasal muosa for allergi rhinitis), in vitro ytokine prodution by PBMC stimulated with antigen has not been w haraterized in patients with allergi diseases. Serist et a/. reported that allergen immunotherapy dereased IL-4 prodution by C4 + T s from allergi individuols." Ohi et a/. reported that both ultured lymphoytes and basophils from the peripheral blood of allergi individuals produed IL-4 in response to ollerqen.!' In the present study, we evaluated the in vitro prodution of IL-4 and -5 by PBMC stimulated with HOM and found that the levels of these ytokines were similar among serum HOM-speifi Ig-positive subjets (groups 1-3). This finding implies that the amount of these ytokines produed in vitro is not related to either the presene or the type of allergi disease. Next we evaluated the orrelation between loal responsiveness and various linial parameters. No orrelation was found between nasal responsiveness and the total serum peripheral blood and Ig level, the eosinophil ount in the IL-4 or -5 prodution by PBMC stimulated with HOM. Bronhial responsiveness (Omin) was negatively orrelated with IL-4 prodution by PBMC among asthma patients (group 1). Although there was a negative orrelation between bronhial responsiveness and IL-5 prodution by PBMC among asthma patients, it was not statistially signifiant. Bronhial responsiveness was also not orrelated with various linial parameters, suh as the total serum Ig level or the eosinophil ount in the peripheral blood. We used methaholine for the assay of bronhial responsiveness and should point out, however, that the possibility exists that the use of histamine for bronhial responsiveness may provide a more signifiant orrelation between bronhial responsiveness and ytokine prodution by PBMC. Robinson et 1. reported a negative orrelation between airway methaholine responsiveness and the number of bronhoalveolar lavage s expressing IL-5 mrna, but not IL-4 mrna24 Akerman et 1. reported that the level of airway hyperresponsiveness in atopi asthma patients was negatively orrelated with the IL-5 immunoreativity of bronhial biopsies, but not with IL-4 immunoretivity" Our results appear to be different from those of Robinson et a/. 24 and Akerman et a/. 25 One reason may be that we assayed in vitro ytokine prodution by PBMC stimulated with antigen, whereas they assayed either bronhoalveolar lavage s or bronhial muosa. However, the signifiant negative orrelation between bronhial hyperresponsiveness and IL-4 prodution, as w as the weaker negative orrelation between bronhial hyperresponsiveness and IL-5 prodution in our asthma patients, supports the notion that the apaity of ytokine prodution by PBMC may reflet bronhial responsiveness among asthmati patients. Finally, there were two non-asthmati subjets who had bronhial hyperresponsiveness in the asthmati range, atopi status (an HOM CAP-RAST sore ~ 3) and an above average prodution of both IL-4 and -5 by peripheral blood (one in the allergi rhinitis group and

7 LOCAL RSPONSIVNSS AND CYTOKINS 51 one with no allergi symptoms). The reason why these subjets did not develop asthma symptoms is unlear and the additional fators neessary to provoke overt symptoms of asthma need to be defined in the future. ACKNOWLDGMNTS We thank all the subjets who volunteered to partiipate in this study. RFRNCS Hargreave F, Ryan G, Thompson NC et a/. Bronhial responsiveness to histamine or methaholine in asthma: Measurement and linial signifiane. J. Allergy C1in. Immuno/. 1981; 68: Borum P. Nasal methaholine hallenge. A test for the measurement of nasal reativity. J. Allergy C1in. Immuno/. 1979; 63: Okuda M. Nasal provoation testing. In: Settipane GA, ed. Rhinitis, 2nd edn. OeanSide Publiations, Providene, 1991; Townley RG, Ryo UY, Kolotkin BM, Kang B. Bronhial sensitivity to methaholine in urrent and former asthmati and allergi rhinitis patients and ontrol subjets. J. Allergy C1in. Immuno/. 1975; 56: Braman SS, Barrows M, DeCotiis BA, Settipane GA, Corrao WM. Airway hyperresponsiveness in allergi rhinitis. A risk fatorfor asthma. Chest 1987; 91: Hagy GW, Settipane GA. Risk fators for developing asthma and allergi rhinitis. J. Allergy C1in. Immuno/. 1976; 58: Ramsdale H, Morris MM, Roberts RS, Hargreave F. Asymptomati bronhial hyperresponsiveness in rhinitis. J. AllergyC1in. Immuno/. 1985; 75: Thompson PJ, Stewart GA. Allergens. In: Holgate S, Churh MK, eds. Allergy. Gower Medial Publishing, London, 1993; Cretios PS. Allergi rhinitis. In: Busse WW, Holgate ST, eds. Asthma and Rhinitis. Blakw Siene, Boston, 1995; Takishima T, Hida W, Sasaki H, Suzuki S, Sasaki T. Diretwriting reorder of the dose-response urves of the airway to methaholine. Chest 1981; 8: Ohi H, Tanaka T, Katada Y et 1. Peripheral blood T lymphoytes and basophils, freshly isolated from housedust-mite-sensitive patients, produe, interleukin-4 in response to allergen-speifi stimulation. Int. Arh. Allergy Immuno/. 1996; 111: Witt C, Stukey MS, Woolok AJ, Dawkins RL. Positive allergy prik tests assoiated with bronhial histamine responsiveness in an unseleted population. J. AllergyC1in. Immuno/. 1986; 77: Woolok AJ, Peat JK, Salome CM et 1. Prevalene of bronhial hyperresponsiveness and asthma in a rural adult population. Thorax 1987; 42: Saito H, Hatake K, Dvorak AM et 1. Seletive differentiation and proliferation of hematopoieti s indued by reombinant human interleukins. Pro. Natl Aad. Si. USA 1988; 85: Del Prete G, Maggi, Parronhi Pet 1. IL-4 is an essential o-fator for the Ig synthesis indued in vitro by human T lones and their supernatants. J. Immuno/. 1988; 14: Liu MC, Hubbard WC, Proud et 1. Immediate and late inflammatory responses to ragweed antigen hallenge of the peripheral airways in allergi asthmatis. Am. Rev. Respir. Dis. 1991; 144: Clutterbuk J, Hirst MA, Sanderson CJ. Human interleukin-5 (IL-5) regulates the prodution of eosinophils in human bone marrow ultures: Comparison and interation with IL-1, IL-3, IL-6, and GM-CSF. Blood 1989; 73: Walsh GM, Hartn A, Wardlaw AJ, Kurihara K, Sanderson CJ, Kay AB. IL-5 enhanes the in vitro adhesion of human eosinophils, but not neutrophils, in a leuoyte integrin (CD 11/18)-dependent manner. Immunology 199; 71: Lopez AF, Sanderson CJ, Gamble JR, Campb HD, Young IG, Vadas MA. Reombinant human interleukin-5 is a seletive ativator of human eosinophil funtion. J. xp. Med. 1988; 167: Bradley BL, AZZQwi M, Jaobson Met 1. osinophils, T lymphoytes, mast s, neutrophils, and marophages in bronhial biopsy speimens from atopi subjets with asthma: Comparison with biopsy speimens from atopi subjets without asthma and normal ontrol subjets and relationship to bronhial hyperresponsiveness. J. Allergy C1in. Immuno/. 1991; 88: Robinson OS, Hamid, Ying Set 1. Predominant Th2-like bronhoalveolar T-lymphoyte population in atopi asthma. N. ngl. J. Med. 1992; 326: Durham SR, Ying S, Varney VA et 1. Cytokine messenger RNA expression for IL-3, IL-4, IL-5, and granuloyte/ marophage-olony-stimulating fator in the nasal muosa after allergen provoation: Relationship to tissue eosinophilia. J. Immuno/. 1992; 148: Serist H, Chelen CJ, Wen Y, Marshall JD, Umetsu DT. Allergen immunotherapy dereases interleukin-4 prodution in CD4 + T s from allergi individuals. J. xp. Med. 1993; 178: Robinson OS, Ying S, Bentley AM et 1. Relationships among numbers of bronhoalveolar lavage s expressing messenger ribonulei aid for ytokines, asthma symptoms, and airway methaholine responsiveness in atopi asthma. J. Allergy C1in. Immuno/. 1993; 92: Akerman V, Marini M, Vittori, Bini A, Vassali G, Mattoli S. Detetion of ytokines and their soures in bronhial biopsy speimens from asthmati patients: Relationship to atopi status, symptoms, and level of airway hyperresponsiveness. Chest 1994; 15: