Serum immunoglobulin E levels predict human airway reactivity in vitro

Serum immunoglobulin E levels predict human airway

reactivity in vitro

D. SCHMIDT, N. WATSON, E. RUEHLMANN, H. MAGNUSSEN* andK. F. RABE

Leiden University Medical Center, Department of Pulmonology, Leiden, The Netherlands and

*Krankenhaus Grosshansdorf, Zentrum fuÈr Pneumologie und Thoraxchirurgie, LVA Hamburg, Grosshansdorf, Germany

Summary

Background Airway hyperresponsiveness to non-speci®c stimuli is one characteristic

feature of airway diseases such as bronchial asthma and chronic bronchitis. Until now,

studies aiming to demonstrate a relationship between in vivo conditions associated with

airway hyperreactivity and in vitro airway responsiveness have been inconclusive.

Objective Since serum immunoglobulin (Ig) E is believed to be one determinant of airway

reactivity in vivo, we studied whether in vitro airway reactivity in lung resection material

from patients with elevated levels of serum IgE was increased as compared with patients

with undetectable IgE. By this approach, we aimed to elucidate the role of circulating IgE

for bronchial smooth muscle reactivity in vitro.

Methods Bronchial rings from nine patients with total serum IgE levels above 200 U/mL

and 10 patients with total serum IgE levels below 10 U/mL were passively sensitized, i.e.

incubated overnight with buffer or sensitizing serum containing high levels of total IgE

(> 250 U/mL). Afterwards, contractile responses to histamine were assessed in the organ

bath.

Results Histamine responsiveness was signi®cantly increased in airways obtained from

patients with IgE levels above 200 U/mL as compared with airways from patients with IgE

levels below 10 U/mL (P< 0.05). Passive sensitization of bronchi from patients with low

IgE signi®cantly increased histamine responsiveness, as compared with non-sensitized

controls from the same patients (P< 0.05). In contrast, passive sensitization of airways from

patients with elevated IgE did not further increase responsiveness. There was no difference

in histamine reactivity between non-passively sensitized and passively sensitized tissue

preparations from patients with IgE above 200 U/mL and passively sensitized tissues from

patients with IgE below 10 U/mL.

Conclusion Our ®ndings reveal that elevated levels of serum IgE predict airway

hyperresponsiveness to histamine in vitro. At the same time, they indicate that the in

vitro model of passive sensitization, in addition to its ability to induce allergen responses,

also mimics conditions of non-speci®c airway hyperreactivity, which are relevant under in

vivo conditions.

Keywords: asthma, bronchial hyperresponsiveness, COPD, human airway smooth muscle,

immunoglobulin E, in vitro, in vivo, sensitization

Clinical and Experimental Allergy, Vol. 30, pp. 233±241. Submitted 26 April 1999; revised

2 July 1999; accepted 27 July 1999.

Introduction

Airway hyperresponsiveness to non-speci®c stimuli is one

characteristic feature of chronic airway diseases such as

bronchial asthma [1] and chronic bronchitis [2±4]. The

Clinical and Experimental Allergy, 2000, Volume 30, pages 233±241

233q 2000 Blackwell Science Ltd

Correspondence: K. F. Rabe, Leiden University Medical Center, Depart-

ment of Pulmonology, C3-P, PO Box 9600, NL-2300 RA Leiden, The

Netherlands (e-mail: [email protected]).

mechanisms underlying bronchial hyperresponsiveness are

not fully understood and are believed to be primarily the

consequence of in¯ammation or changes in smooth muscle

physiology. From in vitro data it appears likely that serum

immunoglobulin (Ig) E plays an important role in the

induction of airway hyperreactivity, since passive sensitiza-

tion of human airwaysÐi.e. the incubation of isolated

bronchi from normal individuals with IgE-rich serumÐ

increases non-speci®c responsiveness to histamine [5,6] and

induces speci®c responsiveness to allergen [7] in vitro. More-

over, circulating speci®c IgE antibodies in vivo cause airway

responsiveness to the corresponding allergen in vitro [8].

However, studies which looked for a relationship between

in vivo parameters and histamine responsiveness of airway

smooth muscle in vitro were inconclusive [9±15]. On the

other hand, epidemiological studies indicate that high levels

of total serum IgE correlate with impaired lung function

[16], increased prevalence of bronchial asthma [17], and

moreover, with bronchial hyperresponsiveness to histamine

in vivo [18]. Until now, however, the role of total serum IgE

levels in vivo for airway responsiveness to histamine in vitro

has not been assessed.

To evaluate the role of circulating IgE for bronchial

smooth muscle reactivity in vitro, we investigated whether

in vitro airway reactivity in lung resection material from

patients with elevated levels of serum IgE was increased as

compared with patients with undetectable IgE. Therefore,

we have analysed, retrospectively, those in vitro experi-

ments on human bronchial ring preparations that have been

performed in our laboratory (a) on airways obtained from

patients with undetectable or with high serum IgE levels, (b)

had passively sensitized and non-sensitized control tissues

and (c) in which histamine concentration±effect curves had

been constructed. Nineteen experiments (10 experiments in

the low-IgE group with total serum IgE levels < 10 U/mL,

nine experiments in the high-IgE group with total serum IgE

levels > 200 U/mL) ful®lled the above criteria and were

selected out of a total of 140 experiments to assess the effect

of high circulating IgE levels on airway responsiveness

in vitro.

Methods

Tissue preparation

Macroscopically normal bronchial tissue was obtained from

19 patients undergoing surgery for lung cancer. Serum IgE

levels of all patients were determined on the day of surgery.

In patients with high total serum IgE levels also speci®c IgE

antibodies against house dust mite were measured.

Immediately after resection, peripheral airways (1±4 mm

internal diameter) were dissected free of alveolar tissue and

cut into rings (2±4 mm length). Tissues were rotated overnight

at room temperature in tubes containing modi®ed Krebs

buffer (composition: NaCl 118.4 mmol/L, KCl 4.7 mmol/L,

MgSO4 0.6 mmol/L, CaCl2 1.3 mmol/L, KH2PO4 1.2 mmol/L,

NaHCO3 25.0 mmol/L, glucose 11.1 mmol/L) in the absence

(non-sensitized controls) or presence of sensitizing serum

(10% v/v). Sensitizing sera were prepared from the whole

blood of 12 individuals demonstrating high total IgE (250 to

> 3000 U/mL) and speci®c antibodies (FAST class $ 3)

against allergen (Dermatophagoides farinae). Sera were

not pooled but were frozen individually in 200±250-mL

aliquots until required.

The next morning bronchial rings were transferred to 10-mL

organ baths containing oxygenated (95% O2, 5% CO2)

modi®ed Krebs buffer (pH 7.4; 37 8C). Tissues were equili-

brated for about 60 min at a resting tension of 300±400 mg,

before the commencement of experimental protocols.

Tension measurements

All responses were recorded as changes in isometric tension

(mg). At the beginning of each experiment a single dose of a

b-adrenoceptor agonist, isoproterenol (1 mmol/L), was applied

to determine the amount of inherent tone. After several

washings and re-equilibration of the tissues, histamine

concentration±effect curves within a range of 10 nmol/L

ÿ 0.3 mmol/L were constructed. In some experiments hista-

mine concentration±effect curves were followed by allergen

(D. farinae) concentration±effect curves within a concentra-

tion range of 0.03±30 U/mL. All concentration±effect

curves were obtained in a cumulative manner, using incre-

mental concentrations spaced at half log10 intervals. At the

end of the experiments, tissues' wet weights were recorded.

Measurements and analysis of results

All concentration±effect curves were acquired in at least

one passively sensitized and one non-sensitized bronchial

ring from the same donor. The potency of histamine was

calculated from concentration±effect curves by non-linear

curve-®tting for each individual tissue and expressed as the

pEC50 value (i.e. ÿlog10 of the concentration of histamine

giving a half maximal effect). Histamine concentration±

effect curves were compared between the different conditions

(passively sensitized and non-sensitized, high and low IgE)

using repeated-measures analysis of variance (ANOVA), with

the different conditions as between±group factor and hista-

mine concentration as within±group factor. In testing for

statistically signi®cant differences between curves, we took

into account the between±group effect (level) as well as its

interaction with the within±group effect (slope). For the

purpose of data presentation, curves were compared with

each other at the different histamine concentrations using

the paired or unpaired t-test, depending on the fact whether

234 D. Schmidt et al.

q 2000 Blackwell Science Ltd, Clinical and Experimental Allergy, 30, 233±241

tissues were paired (sensitized vs non-sensitized) or not paired

(high vs low IgE). Correspondingly, the parameters (maximal

contraction and pEC50) that had been estimated from the

histamine concentration±effect curves were evaluated by t-

tests. The same approach as for histamine was followed in the

analysis of allergen response curves. All values quoted are

mean 6 SEM. The level of statistical signi®cance was assumed

at P� 0.05. t-tests were performed one-tailed regarding

maximal contraction and pEC50 and two-tailed otherwise.

Results

Tissue donors

Experiments on isolated airways from 19 patients were

analysed. Nine patients showed total serum IgE levels

above 200 U/mL (high-IgE group) and 10 patients levels

below 10 U/mL (low-IgE group). In addition, seven patients

of the high IgE group showed speci®c antibodies against

house dust mite (D. farinae) of $ 0.2 U/mL or FAST class

$ 1. In each group, two patients were diagnosed to have

chronic obstructive pulmonary disease (COPD). One patient

in the high-IgE group was known to be sensitized against

house dust mite, but was asymptomatic. One patient in the

low-IgE group had a known bronchial hyperresponsiveness

to histamine (PC20FEV1� 0.2 mg/mL), impaired lung func-

tion (FEV1, 59% of predicted [19]), a negative history of

atopy and a negative skin prick test. All other patients

showed a negative history of atopy and no other severe

pulmonary disease other than lung cancer. Individual char-

acteristics of the patients and mean values are given in Table 1.

Baseline characteristics of bronchial rings

The number of ring preparations obtained from one

patient ranged between 2 and 8 passively sensitized and

Serum IgE and bronchial hyperreactivity 235


Table 1 Individual data of tissue donors

Patient Age Smoking Pack VC FEV1 Total IgE Spec. IgE

no. Sex (years) status years (L) (% pred.) (U/mL) (U/mL)

Total serum IgE levels below 10 U/mL

1 M 67 Ex 70 4.16 134

2 M 62 Ex 100 3.10 86

3 M 74 Yes 50 2.04 56

4 M 58 Yes 90 4.46 80

5 M 76 Yes 60 2.25 55

6 M 69 Ex 40 4.21 92

7 M 64 Ex 80 4.13 80

8 M 62 Ex 48 4.12 92

9 M 66 Yes 25 3.69 99

10 M 72 Yes 45 2.53 59

Mean ± 67 ± 61 3.47 83

6 SEM ± 2 ± 8 0.29 8

Total serum IgE levels above 200 U/mL

11 M 73 Yes 25 3.59 72 3000 0.94

12 M 70 Yes 28 3.79 81 392 0.27

13 F 52 Yes 40 3.69 92 337 0.31

14 F 66 Yes 20 2.56 93 394 0.38

15 F 72 Yes 40 1.96 78 224 0.69

16 M 69 Yes 40 4.52 95 655 0.2

17 M 62 Yes 45 4.23 96 506 0.1

18 M 60 Yes 50 ND ND 296 0.1

19 M 64 Ex 25 5.16 95 309 0.35

Mean ± 65 ± 35 3.68 88 467* 0.29*

6 SEM ± 2 ± 3 0.37 3 (1.3)* (1.3)*

VC� vital capacity. FEV1� forced expiratory volume in one second. % pred.�% predicted [19]. Ex� stopped smoking at least 5 years

ago. Yes� current smoker, smoked till the week before surgery. Spec. IgE� speci®c IgE antibodies against D. farinae, ND� not

determined. *Geometric mean and SEM which is expressed as a factor to be multiplied with the mean value.

1±4 non-sensitized tissues; their mean values sum up to

the given `n'-numbers. The wet weight (11.7 6 0.4 mg,

n� 19), resting tension (344 6 9 mg) and inherent tone

(i.e. the magnitude of relaxation after a single dose of

isoproterenol: 210 6 17 mg) showed no signi®cant differ-

ences either between passively sensitized and non-passively

sensitized tissues or between tissues from high-and low-IgE

donors.

Responses to histamine

Histamine caused concentration-dependent contractions in

both non-passively sensitized and passively sensitized

preparations in the high-IgE and the low-IgE group (Fig. 1,

Table 2).

The concentration±effect curves obtained in non- passively

sensitized tissues differed signi®cantly between both groups

(ANOVA, P� 0.002). Speci®cally, maximal contractions to

histamine were increased in non-sensitized preparations

from patients with IgE levels above 200 U/mL, as compared

with non-sensitized airways from patients with IgE levels

below 10 U/mL (P� 0.03). The difference in potency of

histamine between both groups did not reach statistical

signi®cance (P� 0.13).

In the low-IgE group, histamine concentration±effect

curves were signi®cantly different between sensitized and

non-sensitized tissues (ANOVA, P < 0.001). Passive sensitiza-

tion of bronchi from patients with low IgE signi®cantly

increased both the potency of histamine (P� 0.017) and

maximal contractions (P� 0.003) obtained, as compared

with non-sensitized tissues from the same individuals

(Table 2). In contrast, in the high-IgE group, histamine

concentration±effect curves did not differ signi®cantly

between passively sensitized and non-sensitized tissues

(ANOVA, not signi®cant; Table 2 and Fig. 1). Correspond-

ingly, maximal contraction and pEC50 were not different.

Furthermore, concentration±effect curves of both pas-

sively sensitized and non-passively sensitized tissues from

the high-IgE group were almost identical to those of

passively sensitized tissues in the low-IgE group (ANOVA,

not signi®cant; Fig. 1); there was no signi®cant difference in

potency or maximal contractions (Table 2).



800

600

400

200

0

Co

ntr

acti

on

(m

g)

[Histamine] (mol/L)

*

*

*

*

**

(a)

10–8 10–7 10–6 10–5 10–4 10–3

800

600

400

200

0

[Histamine] (mol/L)

(b)

10–8 10–7 10–6 10–5 10–4 10–3

Fig. 1. Contractile concentration±effect curves to histamine of human bronchi from donors with total serum IgE (a)< 10 U/mL (circles) and

(b)> 200 U/mL (squares) in non-sensitized (open) and passively sensitized (closed) preparations. Contractions were expressed as mg

changes in tension. Data are the mean 6 SEM of 10 experiments in the low-IgE and of nine experiments in the high-IgE group. *P< 0.05,

regarding contractile responses to individual concentrations of histamine

Responses to allergen

Allergen (D. farinae) caused concentration-dependent con-

tractions of passively sensitized tissues in the low-IgE

group, but not in non-sensitized control tissues from the

same donors; this difference was statistically signi®cant

(ANOVA, P < 0.001, n� 6; Fig. 2). In contrast, isolated air-

ways from donors of the high-IgE group who showed

speci®c antibodies against house dust mite demonstrated

allergen-induced contractions in both passively sensitized

and non-passively sensitized tissues. The concentration

effect-curves differed signi®cantly from each other (ANOVA,

P� 0.003) and maximal contractions in non-passively sen-

sitized tissues were about 50% of those obtained in sensi-

tized tissues (P� 0.04, n� 5; Fig. 2).

Concentration-response curves in non-passively sensi-

tized tissues from the high-IgE group were signi®cantly

different from the responses obtained in the non-sensitized

tissues of the low-IgE group (ANOVA, P� 0.002; Fig. 2 and

Table 2). Correspondingly, maximal contractions were

elevated (P� 0.02).

Relationship between smoking status and

bronchial reactivity

As the smoking status differed between the low and the

high-IgE group, we additionally analysed our data with

respect to the potential effect of current vs ex-smoking.

There were 5/5 current/ex-smokers in the low-IgE and 8/1 in

the high-IgE group, with a lower number of pack years in

the latter group. In passively sensitized tissues of the low-

IgE group, maximal contraction was signi®cantly higher in

the current smokers than in the ex-smokers (P� 0.04). The

same was true in non-sensitized tissues (P� 0.02). How-

ever, there was no difference in pEC50 values, neither

between sensitized nor between non-sensitized tissues of

the current and ex-smokers from the low-IgE group. As the

high-IgE group comprised 8/9 current smokers, the results

of passive sensitization in current smokers did not differ

from those of the whole group, in which passive sensitization

led to a further increase in smooth muscle responsiveness.

Discussion

Our study demonstrates that total serum IgE is an in vivo

determinant for airway smooth muscle hyperreactivity in

vitro. Isolated airways from patients with high levels of total

serum IgE were hyperresponsive to histamine in vitro as

compared with airways from patients with low IgE levels.

Furthermore, passive sensitization of airways from patients

with elevated IgE levels did not lead to a further increase in

histamine responsiveness. These results underline the

important role of serum IgE for bronchial hyperreactivity

and prove passive sensitization to be a useful in vitro model

for non-speci®c airway hyperresponsiveness.

The approach which we used is based on previous reports

that human airway smooth muscle preparations obtained

from nonatopic individuals become hyperresponsive to

histamine after incubation with IgE-rich serum, which is

re¯ected in an increase in maximal contraction and a

leftward shift of the concentration±effect curve of approxi-

mately a half log concentration [5±7]. Therefore, we hypoth-

esized that patients whose airways had already been



Table 2. Airway responses to histamine

and allergen in vitro Max. contr. Max. contr.

pEC50 histamine D. farinae

Group histamine (mg) (% hist. max.)

Low-IgE (< 10 U/mL)

Non-sensitized 5.10 6 0.13 496 6 56 11 6 10

Passively sensitized 5.59 6 0.15* 710 6 70* 79 6 8*

High-IgE (> 200 U/mL)

Non(passively) sensitized 5.37 6 0.19 689 6 84* 45 6 8*

Passively sensitized 5.49 6 0.14* 707 6 73* 80 6 15*#

Potency of histamine and maximal contractions to histamine and allergen (D. farinae) in

passively sensitized and non-sensitized bronchial rings from donors with serum IgE levels of

< 10 U/mL and > 200 U/mL. The potency of histamine is expressed as the pEC50 value, i.e.

ÿlog10 of the concentration of histamine giving a half maximal effect. Maximal tension

changes after the highest concentration of histamine (0.3 mmol/L) were measured in mg.

Contractile responses to allergen (30 U/mL) were expressed in percent of maximal responses

to histamine. Data are given as mean values 6 SEM. * P< 0.05, in comparison with the non-

sensitized tissues from the low-IgE group. #P < 0.05, in comparison with the non-sensitized

tissues from the high-IgE group.

exposed to high levels of serum IgE in vivo should demon-

strate airway hyperresponsiveness in vitro as compared with

those with undetectable serum IgE levels. Indeed, our data

revealed that smooth muscle preparations from patients with

high serum IgE levels showed a signi®cant increase in

concentration±effect curves and maximal contractions to

histamine, neither of which could be further enhanced by

passive sensitization. Furthermore, histamine responsive-

ness of non-passively sensitized airways from patients with

high serum IgE levels was not different from that observed

after passive sensitization of airways taken from patients

with undetectable IgE levels. These ®ndings con®rm our

hypothesis that the patients' serum, which contained high

levels of total IgE, had already induced elevated or even

`maximal' non-speci®c responsiveness of airway smooth

muscle before resection.

These in vitro results ®t well with the positive correlation

between serum IgE levels and bronchial responsiveness in

vivo, which has been demonstrated in epidemiological

studies in both adults and children [18,20±23]. Serum IgE

level appear to be a marker of the immune responses that

result in the development of airway hyperresponsiveness

[24] or bronchial asthma [25]; speci®cally, levels above

200 U/mL have been shown to be associated with airway

hyperresponsiveness in vivo, independently of skin prick

test reactions [22]. Our study demonstrates that circulating

IgE levels are also predictors of non-speci®c bronchial

responsiveness ex vivo.

The validity of our data is supported by the fact that

passive sensitization with serum containing speci®c IgE

antibodies induces airway responsiveness to the correspond-

ing allergen, as known [7], and are consistent with previous

reports that bronchi from patients with high speci®c IgE

antibodies contract in vitro when exposed to the correspond-

ing allergen [8,26±28]. Interestingly, passive sensitization

with serum containing higher concentrations of speci®c

antibodies than that of tissue donors even further increased

allergen-induced contraction in vitro.

It is known that the induction of allergen responsiveness

through passive sensitization is dependent on speci®c IgE,



100

80

60

40

20

0

Co

ntr

acti

on

(%

his

tam

ine

max

.)

[D. farinae] (U/mL)

(a)

0.01 0.1 1 10 100

*

100

80

60

40

20

0

[D. farinae] (U/mL)

(b)

0.01 0.1 1 10 100

*

**

Fig. 2. Contractile concentration±effect curves to D. farinae of human bronchi from donors with total serum IgE (a) < 10 U/mL (circles)

and (b)> 200 U/mL and speci®c IgE antibodies against D. farinae (squares), in non-sensitized (open) and passively sensitized (closed)

preparations. Contractions were expressed as percentage of maximal contraction to histamine. Data are the mean 6 SEM of six experiments

in the low-IgE and of ®ve experiments in the high-IgE group. *P< 0.05, in comparison with the non-sensitized tissues from the low-IgE group

regarding maximal contractile responses. **P< 0.05, in comparison with the non-sensitized tissues from the high-IgE group regarding maximal

contractile responses

since complexing of IgE by anti-IgE antibodies nearly

abolishes allergen-induced responses, through inhibition

of IgE coupling to Fc receptors on mast cells [29]. In

contrast with allergen responsiveness, however, the increase

in histamine responsiveness after passive sensitization could

be inhibited neither by pretreatment with anti-IgE antibodies

[29] nor by depletion of serum IgE [30]. Conversely, incuba-

tion with IgE alone was not suf®cient to increase non-speci®c

responsiveness to histamine in vitro [7]. Therefore, IgE-rich

sera must contain a so far unknown factor that is closely

linked both to IgE levels and to the ability of serum to induce

histamine hyperresponsiveness [7]. Despite that, however, our

data indicate that serum IgE could serve as a surrogate marker

of the immunological processes that are linked to the devel-

opment of non-speci®c airway hyperresponsiveness.

From epidemiological studies it is known that not only

atopy but also smoking can be associated with elevated total

serum IgE levels [31,32]. The observation that high IgE

levels are related to current smoking and normalize after

cessation of smoking [33±35] is consistent with our results.

Furthermore, our data re¯ect the in vivo observations that (a)

total serum IgE levels are associated with current smoking

and (b) correlate with bronchial hyperresponsiveness. In

addition, among nonatopic individuals, bronchial respon-

siveness occurs more frequently in current smokers as

compared with non- or ex-smokers; however, in atopic

subjects smoking does not lead to increased prevalence of

airway responsiveness [36]. Although the relationship

between current smoking and elevated levels of total IgE,

as observed in epidemiological studies, is not fully under-

stood [36,37], stimulation of TH2-type (interleukin-4) cyto-

kine production by smoking may represent one mechanistic

link [38±42]. Thereby, current smoking may be associated

with the unknown serum factor that is linked to total IgE and

causes non-speci®c airway responsiveness [7]. It is impor-

tant to note that airway responsiveness in vitro could be

enhanced by passive sensitization only in current smokers

who showed low levels of circulating IgE, whereas in

smokers with high IgE levels airway responsiveness in

vitro could not be further increased. Although it is dif®cult

to exclude smoking as a confounding factor in our study,

this association between IgE levels and inducibility of

airway hyperresponsiveness in vitro renders it unlikely

that smoking was the primary cause of airway hyperrespon-

siveness. Therefore, the association between smoking and

total IgE as observed in our study is consistent with our

notion that it is the elevated IgE or rather the `unknown

factor' level in vivo which is associated with airway hyper-

responsiveness in vitro.

Previous studies which investigated the relationship

between in vivo airway responsiveness, as determined by

histamine inhalation challenges, and in vitro responsiveness

of isolated airways, as determined by the potency of

histamine (EC50), have been inconclusive [10±15] and

still it remains unresolved to what extent bronchial hyper-

responsiveness in vivo results from in¯ammation, neural

dysregulation, or intrinsic abnormality of airway smooth

muscle. In vitro data obtained with passive sensitization

demonstrate that airway hyperresponsiveness can be

induced and modulated in isolated bronchial rings and that

there is a close analogy between in vivo and in vitro

®ndings. Therefore, owing to the fact that the model of

passive sensitization mimics important characteristics of

airway hyperresponsiveness in vivo, it is rather unlikely

that airway hyperresponsiveness in vivo results only from

in¯ammation or neural dysregulation. In contrast, the rela-

tionship between airway hyperresponsiveness to histamine

in vitro and exposure to IgE-rich serum in vivo is likely to

re¯ect an abnormality of airway smooth muscle that is

essential in the pathogenesis of airway hyperreactivity.

In summary, our study demonstrates that airways from

patients with high levels of circulating IgE are hyperreactive

to histamine in vitro. Therefore, total serum IgE is a

predictor for non-speci®c airway hyperreactivity in vitro,

in accordance with epidemiological studies, which show a

link between serum IgE levels and bronchial hyperrespon-

siveness in vivo. Thus, it is likely that serum IgE or closely

associated factors play a crucial role in the development of

bronchial hyperreactivity under both in vivo and in vitro

conditions. Since passive sensitization appears to be a

model that closely mimics in vivo conditions, it remains to

be seen, for example, whether therapeutic intervention with

anti-IgE antibodies will not only alter allergen responses

[43] but also non-speci®c hyperresponsiveness which could

be of even greater relevance for the treatment of bronchial

asthma.

Acknowledgements

The authors would like to thank the surgical staff of the

Krankenhaus Groûhansdorf for their co-operation and the

members of the Clinical Laboratory for performing the total

and speci®c IgE analysis. We are grateful to Rudolf JoÈrres,

Krankenhaus Groûhansdorf, for valuable comments to the

manuscript and statistical advice. This work was supported

by a grant from GlaxoWellcome Medicines Research (UK).

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Serum immunoglobulin E levels predict human airway reactivity in vitro