J Appl Physiol

91: 1029–1034, 2001.

Inflammatory and mechanical factors of allergen-

induced bronchoconstriction in mild asthma and rhinitis
EMANUELE CRIMI, MANLIO MILANESE, SUSANNA ODDERA, CARLO MEREU,
GIOVANNI A. ROSSI, ANNAMARIA RICCIO, G. WALTER CANONICA, AND VITO BRUSASCO
Dipartimenti di Scienze Motorie e Riabilitative di Medicina Interna
e di Oncologia Biologica e Genetica, Università di Genova, 16132 Genova;
and Divisione di Pneumologia, Istituto G. Gaslini, 16130 Genova, Italy
Received 29 November 2000; accepted in final form 9 April 2001

Crimi, Emanuele, Manlio Milanese, Susanna Od-
dera, Carlo Mereu, Giovanni A. Rossi, Annamaria
Riccio, G. Walter Canonica, and Vito Brusasco. Inflam-
matory and mechanical factors of allergen-induced broncho-
constriction in mild asthma and rhinitis. J Appl Physiol 91:
1029–1034, 2001.—We studied whether different bronchial
responses to allergen in asthma and rhinitis are associated
with different bronchial inflammation and remodeling or
airway mechanics. Nine subjects with mild asthma and eight
with rhinitis alone underwent methacholine and allergen
inhalation challenges. The latter was preceded and followed
by bronchoalveolar lavage and bronchial biopsy. The re-
sponse to methacholine was positive in all asthmatic but in
only two rhinitic subjects. The response to allergen was
positive in all asthmatic and most, i.e., five, rhinitic subjects.
No significant differences between groups were found in
airway inflammatory cells or basement membrane thickness
either at baseline or after allergen. The ability of deep inha-
lation to dilate methacholine-constricted airways was greater
in rhinitis than in asthma, but it was progressively reduced
in rhinitis during allergen challenge. We conclude that 1)
rhinitic subjects may develop similar airway inflammation
and remodeling as the asthmatic subjects do and 2) the
difference in bronchial response to allergen between asthma
and rhinitis is associated with different airway mechanics.
airway hyperresponsiveness; basement membrane; bron-
choalveolar lavage; bronchial biopsy; deep inhalation
IT HAS BEEN RECENTLY SUGGESTED that asthma and rhini-
tis represent two different phenotypes of the same
disease (9). In support of this hypothesis are the obser-
vations that experimental exposure to allergen causes
bronchial narrowing (2, 6) and inflammation (17) in
subjects suffering from rhinitis alone, i.e., without
asthma. An important difference between asthma and
rhinitis alone is the much greater dose of allergen
necessary to cause significant bronchoconstriction in
the latter (2), which may explain why some allergic
subjects develop asthma symptoms on natural expo-
sure to allergen, whereas others do not. The reasons for
a greater bronchial responsiveness to allergen in
asthma than in rhinitis are, however, still largely un-
explained.
In subjects with a similar type and degree of sys-
temic allergic sensitization, differences in bronchial
responsiveness to allergen may be related to differ-
ences in bronchial inflammation and remodeling or
airway mechanics. It has been previously shown that
the bronchi of rhinitic subjects are susceptible to de-
velop an inflammatory response to allergen that is
indistinguishable from that of asthmatic bronchi (17).
However, only bronchoalveolar lavage (BAL) was used,
which precluded any evaluation of bronchial wall in-
flammation or remodeling. Furthermore, possible dif-
ferences of airway mechanics in response to allergen
between asthmatic and rhinitic subjects were not in-
vestigated.
In the present study, we used both BAL and bron-
chial biopsy (BB) to evaluate bronchial inflammation
and remodeling in subjects with mild asthma (with or
without rhinitis) and subjects with rhinitis alone. Pos-
sible differences in airway mechanics between the two
groups were investigated by comparing 1) the bron-
chial responsiveness to allergen and to methacholine
(MCh), and 2) the effects of deep inhalation (DI) on
airway caliber (15) during allergen and MCh chal-
lenges.
METHODS
Subjects. The study was conducted on 17 male, nonsmok-
ing subjects, who were sensitized to house dust mite (HDM),
as documented by a third-to-fourth radioallergosorbent test
class (Pharmacia, Uppsala, Sweden) and a positive skin prick
test. The latter was defined by a wheal response to HDM
extract (Lofarma, Milan, Italy) equal to or larger than that
caused by 10 mg/ml histamine. Eight subjects (age 25 ⫾ 3 yr)
had a history of rhinitis and never experienced symptoms
that may be related to asthma or airway hyperresponsive-
ness, such as wheeze, waking up at night with shortness of
breath or chest tightness, or cough or sputum on exposure to
house dust (4). Nine subjects (age 24 ⫾ 3 yr) had bronchial
asthma, according to the definition of the American Thoracic
Society (1). At the time of the study, all subjects were in a

Address for reprint requests and other correspondence: V. Bru- The costs of publication of this article were defrayed in part by the
sasco; Dipartimento di Scienze Motorie e Riabilitative; Università di payment of page charges. The article must therefore be hereby
Genova; Largo R. Benzi, 10:16132 Genova, Italy (E-mail: brusasco marked ‘‘advertisement’’ in accordance with 18 U.S.C. Section 1734
@dism.unige.it). solely to indicate this fact.

http://www.jap.org 8750-7587/01 $5.00 Copyright © 2001 the American Physiological Society 1029

Downloaded from www.physiology.org/journal/jappl by ${individualUser.givenNames} ${individualUser.surname} (185.050.250.187) on December 26, 2018.

 1030 BRONCHIAL RESPONSE TO ALLERGEN IN RHINITIS AND ASTHMA
stable clinical condition, with a 1-s forced expiratory volume
(FEV1) ⬎70% of predicted (16), and none of them was taking
inhaled or oral steroids, cromolyn, antihistamine, or regular
bronchodilators. Asthmatic subjects were using short-acting
␤2-agonists on demand, which were avoided for 12 h before
studies. The study was approved by the institutional ethics
committee, and subjects gave written, informed consent.
Protocol. All subjects first underwent a MCh inhalation
challenge followed, 24 h later, by BAL and BB. After 1 wk at
least, they underwent a HDM inhalation challenge followed,
24 h later, by BAL and BB. Six asthmatic and six rhinitic
subjects attended the laboratory on two further occasions to
have MCh and HDM challenges repeated to determine the
effects of DI.
Lung function measurements. A Vmax 6200 system (Sen-
sorMedics, Yorba Linda, CA) was used for baseline pulmo-
nary function tests and MCh challenge. Portable spirometers
(Micro Medical, Rochester, Kent, UK) were used to monitor
lung function during the allergen challenge, with the same
subject using the same instrument at the hospital and at
home. On each occasion, the highest of three FEV1 measure-
ments within 100 ml was retained. The effects of DI during
allergen and MCh challenges were determined by comparing
expiratory flows at 40% of control forced vital capacity (FVC)
during forced expiratory maneuvers started from full infla-
tion (maximal flow) and from ⬃60% of FVC (partial flow)
using dedicated software (Vmax 6200, SensorMedics). Three
acceptable sets of partial and maximal flow-volume curves
were obtained at control, and one at each step of the chal-
lenges. Each set consisted of a forceful expiration from ⬃60%
of FVC to residual volume, followed by a fast inhalation to
total lung capacity and, without breath hold, a forceful expi-
ration to residual volume. The bronchodilator effect of DI was
inferred from the slope (MP slope) and the intercept (MP
intercept) of the regression line of maximal (M) vs. partial (P)
flows measured at all steps of the challenges (15). In this
analysis, a slope ⬍1 indicates that, for any given decrease of
partial flow, the maximal flow decreases less, thus suggest-
ing a bronchodilator effect of DI. The coefficient of variation
(CV) of MP slope and MP intercept measurements was de-
termined in a separate group of stable asthmatic subjects to
be 14 and 20%, respectively (unpublished data).
MCh challenge. Solutions of MCh were prepared by adding
distilled water to dry powder MCh chloride (Laboratorio
Farmaceutico Lofarma). Aerosols were delivered by an SM-1
Rosenthal breath-activated dosimeter (SensorMedics) driven
by compressed air (30 psi) with 1-s actuations. The aerosol
output at the mouth was 10 ␮l per actuation. Aerosols were
inhaled during quiet tidal breathing. After 20 inhalations of
saline as a control, the subjects inhaled double-increasing
doses of MCh from 0.02 mg until FEV1, measured 1 min after
each dose, decreased below 80% of control value or the max-
imum dose of 1.2 mg was achieved. The double increments of
dose were obtained by using two MCh concentrations (1 and
10 mg/ml) with appropriate numbers of breaths. A 3-min
interval was allowed between dose increments. The noncu-
mulative dose causing a 20% reduction of FEV1 from control
(PD20) was calculated by interpolation between two adjacent
points of the log dose-response curve. In subjects who did not
reach a 20% reduction of FEV1, the last MCh dose was
retained as PD20 for statistical analysis. PD20 values ⬍1.2
mg were considered positive for airway hyperresponsiveness.
HDM challenge. A powder-allergen method was used (13).
The HDM extract consisted of a Dermatophagoides pteronys-
sinus and farinae dry mix (50:50) predosed in arbitrary units
(AU) by radioallergosorbent test inhibition technique with an
internal (Laboratorio Farmaceutico Lofarma) reference ex-
J Appl Physiol • VOL
Downloaded from www.physiology.org/journal/jappl by ${individualUser.givenNames} ${individualUser.surname} (185.050.250.187) on December 26, 2018.
tract to which a value of 1,000 AU had been assigned. One
arbitrary unit corresponded to 7.5 IU of the World Health
Organization International Union of Immunological Societ-
ies Dermatophagoides pteronyssinus International Standard
(National Institute for Biological Standards and Control code
82/518). The mean particle size of the allergen powder deter-
mined by laser particle sizer (Analysette 22, Fritsch, Idar,
Oberstain, Germany), with ethanol as dispersing fluid, was
2.5 ␮m, with 99.9% of the particles ⬍ 18.2 ␮m, 70% ⬍ 3.9 ␮m,
50% ⬍ 2.5 ␮m, 30% ⬍ 1.5 ␮m, and 10% ⬍ 0.7 ␮m. Micronized
allergen in the form of inert lactose powder was contained in
rigid gelatin capsules (40 ⫾ 2 mg of powder each). The
following doses were made available by the manufacturing
firm: 5, 10, 25, 50, 100, 200, and 400 AU. The powder was
administered through a turboinhaler device (Schiapparelli-
Searle, Torino, Italy) with a series of slow inspiratory capac-
ity maneuvers repeated until the capsule was empty. A
control measurement of FEV1 was obtained 15 min after
inhalation of lactose. The challenge was then started from
the lowest dose of allergen (5 AU) and stopped when the
FEV1, measured 15 min after the dose, was decreased below
80% of control value. If such a decrease of FEV1 did not occur
with the dose of 790 AU, an additional 400 AU were admin-
istered, and the challenge stopped at the cumulative dose of
1,190 AU. In rhinitic subjects, the first three doses (5, 10, and
25 AU) were inhaled consecutively as a unique dose of 40 AU.
Cumulative PD20 was calculated by interpolation of the dose-
response curve. In subjects who did not reach a 20% reduc-
tion of FEV1, the last allergen dose was retained as PD20 for
statistical analysis. FEV1 measurements were then taken
hourly for 24 h, except when the subject was asleep. A
decrease of FEV1ⱖ 20% of control value recorded at any time
within 24 h after allergen inhalation was considered as
evidence of a positive response.
Bronchoscopic procedure. After premedication with atro-
pine (0.5 mg im) and prometazine hydrochloride (50 mg im),
lidocaine (2% solution) and adrenaline (0.1:1,000 solution)
were instilled into the nostrils. A fiber-optic bronchoscope
(Olympus BF, type 1T20) was then passed through the nose
or the mouth without endotracheal tube and, after local
anesthesia of pharynx and airways, wedged into a subseg-
mental bronchus of the right middle lobe. Sterile saline (5
aliquots of 20 ml each) was instilled. The BAL fluid was
collected by applying a negative pressure (50–120 mmHg) at
the proximal port of the bronchoscope. Immediately after
BAL, four biopsy specimens were taken from the right upper
lobe distal bifurcation.
BAL analysis. After filtration through two layers of sterile
gauze, the fluid was centrifuged at 500 rpm for 5 min. The
cell pellet was washed once and resuspended in Hanks’
balanced salt solution, without Ca2⫹ and Mg2⫹, at a concen-
tration of 106 cells/ml. A small sample of the cell suspension
was centrifuged (Cytospin, Shandon Southern Instruments,
Sewickley, PA) at 500 rpm for 5 min, spinning ⬃100,000 cells
onto a glass slide. Cells were air dried and stained (Diff-Quik;
Merz & Dade, Dudingen, Switzerland) for a differential cell
count of 300 cells per slide, by light microscopy. Epithelial
cells were not included in the differential count, and their
absolute values are reported separately. Cell-free superna-
tants were assayed for eosinophil cationic protein (ECP) by
fluoroimmunoassay (Pharmacia UniCAP System Fluoroen-
zymeimmunoassay, Pharmacia Diagnostic) and for tumor
necrosis factor-␣ (TNF-␣) content by enzyme immunoassay
(Cytelisa, Cytimmune Science, College Park, MD) following
the manufacturer’s instruction. The sensitivity of ECP and
TNF-␣ assay is 2 ng/ml and 4.8 pg/ml, respectively. The
albumin concentration in the supernatant was determined
91 • SEPTEMBER 2001 • www.jap.org
 BRONCHIAL RESPONSE TO ALLERGEN IN RHINITIS AND ASTHMA 1031

Table 1. Bronchoalveolar lavage data

Rhinitis (n ⫽ 8) Asthma (n ⫽ 9)

Pre-HDM Post-HDM Pre-HDM Post-HDM

Cells ⫻ 10 6
2 (1.2–4.1) 3.65 (2.08–8)* 1.17 (0.94–2.5) 3.55 (3.07–8.1)
Epithelial cells ⫻ 104 17.2 (7.5–30) 10.3 (8.5–19) 11.8 (3.45–25) 9.5 (2.85–25.3)
Macrophages, % 92.8 (89.1–96.4) 84.6 (81.1–89.7)* 90.5 (87.5–93.4) 84.2 (71.8–90)*
Lymphocytes, % 4.3 (1.5–6.5) 1.4 (0.5–10.3) 2.7 (2–8.3) 4 (2.1–12.1)
Neutrophils, % 0.75 (0–1) 2 (0.5–2.6) 1.1 (0–1.3) 1.3 (0.9–2.5)
Eosinophils, % 1.3 (0.7–5.5) 8.7 (5.2–3.6)* 4.4 (1.8–5.1) 6.6 (3.5–8.8)
Albumin, mg/100 ml 8.1 (5.2–11.8) 6.8 (5.6–11.9) 7 (6–8.3) 8.6 (6.5–10)
ECP, pg/ml 3.0 (1.4–16.5) 25.8 (7.5–47.4)* 3.1 (2–8.8) 16.1 (8.2–23)*
TNF-␣, pg/ml 121 (56.5–180) 80 (59–99) 81 (78–140) 70 (61–78)
IgE, U/ml
Total (n ⫽ 5) 0.58 (0.51–1.02) 0.83 (0.77–1.12)
Specific (n ⫽ 5) 0.44 (0.35–0.85) 0.8 (0.41–1.22)
Values are medians with lower and upper quartiles in parentheses; n, no. of subjects. HDM, house dust mite; ECP, eosinophil cationic
protein; TNF-␣, tumor necrosis factor-␣. * P ⬍ 0.05 vs. pre-HDM.

by nephelometry. Total and specific IgE to dermatophagoides
were measured by reverse enzyme allergosorbent test (Labo-
ratorio Farmaceutico, Lofarma), which is based on the cap-
ture of both total and specific IgE by a specific antihuman IgE
antibody and the subsequent reaction with streptoavidin-
peroxidase and chromogenic substrate (8). The optical den-
sity was read on a human IgE reference curve titrated refer-
ring to the Second World Health Organization IgE standard
75/502. For both total and specific IgE, the lower and upper
detection limits were 0.2 and 100 U/ml, respectively.
BB analysis. Biopsy specimens were fixed in 10% formal-
dehyde at 4°C for 4 h, embedded in paraffin, cut at 5 ␮m with
a rotative microtome, and stained with hematoxylin and
eosin and toluidine blue. Eight too small and incorrectly
oriented biopsies were discarded, but for each subject at least
one biopsy was available for analysis. Microscopic examina-
tion was performed by one independent observer, who was
unaware of the aim of the study. For cell counts (eosinophils
and mast cells), bronchial mucosa was examined to a depth of
100 ␮m from the basement membrane over adjacent non-
overlapping high-power fields (at ⫻500 magnification with
the aid of an eyepiece graticule) until all of the available area
was covered. For each quantification, three sections were
examined, and cells were expressed as number per square
millimeter of submucosa. Mast cells were also examined at
⫻1,000 magnification to detect ongoing degranulation (gran-
ules surrounding the cell or crossing the cell membrane). The
intraobserver mean CV for three repeated measurements
was 9% for eosinophils and 8% for mast cells. The thickness
of reticular basement membrane (RBM) was measured on
hematoxylin- and eosin-stained preparations by light micro-
scope image analysis (Q 500, Leica, Cambridge, UK) at ⫻400
magnification from the base of bronchial epithelium to the
outer limit of the reticular lamina, at regular intervals (20
␮m), until 40 readings were obtained. The intraobserver
mean CV for three replicate measurements was 3%. The
percentage of RBM covered by epithelium was also calcu-
lated.
Statistical analysis. Unpaired t-test was used to compare
anthropometric and lung function data between groups.
Fisher’s exact test was used to compare categorical variables.
Two-factor, repeated-measure ANOVA with least significant
difference post hoc test was used to compare MP slopes and
MP intercepts between and within groups. Mann-Whitney
U-test was used for comparisons of BAL and BB data be-
tween groups before and after allergen challenge. Wilcoxon
matched-rank test was used to compare data before and after
allergen challenge. P ⬍ 0.05 was considered statistically
significant. Data are presented as means ⫾ SD or medians
with interquartile ranges.
RESULTS
BAL data. See Table 1. Neither at baseline nor after
HDM challenge were the differences in inflammatory
cell counts and epithelial cells statistically significant
between asthma and rhinitis (P ⬎ 0.2, for all compar-
isons). Total cell number and percentages of eosino-
phils were increased after HDM challenge significantly
in rhinitis and nonsignificantly in asthma, whereas the
percentage of macrophages decreased in both groups.
The ECP and TNF-␣ contents of supernatant were not
significantly different at baseline (P ⬎ 0.2), and ECP
increased similarly in the two groups after HDM chal-
lenge. Total and specific IgE, detectable in five rhinitic
and five asthmatic subjects, did not show significant
differences between groups (P ⬎ 0.3). Also the albumin

Table 2. Bronchial biopsy data

Rhinitis (n ⫽ 8) Asthma (n ⫽ 9)

Pre-HDM Post-HDM Pre-HDM Post-HDM

Eosinophils, mm⫺2 20.5 (8.5–61) 68.5 (31–110)* 17 (8–35) 17 (15–24)

Mast cells, mm⫺2 28.5 (28–42.5) 42.5 (21.5–61.5) 29 (10–38) 38 (19–57)
Degranulating mast cells, % 33 (23.5–67.5) 51 (33–74) 33 (0–66) 62 (40–75)*
RBM thickness, ␮m 10 (7.4–15.6) 11.2 (9.1–11.6) 12 (8.8–14.1) 10.6 (6.8–16)
Epithelium-covered RBM, % 85 (68.5–100) 87.5 (76–96) 78 (62–100) 75 (36–80)
Values are medians with lower and upper quartiles in parentheses; n, no. of subjects. * P ⬍ 0.05 vs. preallergen. RBM, reticular basement
membrane.

J Appl Physiol • VOL 91 • SEPTEMBER 2001 • www.jap.org

Downloaded from www.physiology.org/journal/jappl by ${individualUser.givenNames} ${individualUser.surname} (185.050.250.187) on December 26, 2018.

 1032 BRONCHIAL RESPONSE TO ALLERGEN IN RHINITIS AND ASTHMA

content of BAL supernatant was not significantly dif-

ferent between groups (P ⬎ 0.5).
BB data. See Table 2. There were no significant
differences in inflammatory cell numbers between rhi-
nitis and asthma at baseline (P ⬎ 0.5 for all compari-
sons), but the number of eosinophils increased signifi-
cantly in rhinitis, and the percentage of degranulating
mast cells increased in asthma after HDM. Neither
RBM thickness nor the percentage of RBM covered by
intact epithelium were significantly different between
groups (P ⬎ 0.7 for both comparisons), although the
latter tended (P ⫽ 0.06) to be less in asthma after
HDM.
Lung function data. See Table 3. Both FEV1 and its
ratio to FVC (FEV1/FVC) were less in asthma than in
rhinitis, suggesting mild airway obstruction at base-
line in the former. Individual baseline FEV1 values on
the allergen challenge days were always within ⫾8% of
those on the MCh challenge days, without significant
differences between or within groups. Both allergen
PD20 and MCh PD20 were significantly less in asthma
than in rhinitis. The response to MCh was positive in
all asthmatic subjects but in only two rhinitic subjects;
this difference was statistically significant (P ⬍ 0.01).
The response to HDM was positive in all asthmatic
subjects and in five out of eight rhinitic subjects, a
difference that was not statistically significant (P ⬎
0.08). Of the two rhinitic subjects with positive re-
sponse to MCh, one (PD20 ⫽ 1,200 ␮g) did respond to
HDM, whereas the other (PD20 ⫽ 487 ␮g) did not. The
late-phase bronchial response to HDM tended to be
more severe in asthma than in rhinitis (maximum late
FEV1 fall: 32 ⫾ 16 vs. 18 ⫾ 11%, P ⫽ 0.05), although
five asthmatic subjects were given bronchodilator
treatment to relieve symptoms.
In both groups, DI had a bronchodilator effect during
induced bronchoconstriction, as indicated by the MP
slope values that were significantly (P ⬍ 0.01) less than
unity (Fig. 1). This effect was greater in rhinitis than in
Table 3. Lung function data
Rhinitis (n ⫽ 8) Asthma (n ⫽ 9)
FEV1, % predicted 104 ⫾ 8 91 ⫾ 10†
FEV1/FVC, % 86 ⫾ 6 77 ⫾ 7*
Log MCh-PD20, mg 3.031 ⫾ 0.139 2.078 ⫾ 0.400‡
Log HDM-PD20, AU 2.940 ⫾ 0.184 2.446 ⫾ 0.551*
MCh positive/negative, no. 2/6 9/0†
HDM positive/negative, no. 5/3 9/0
MP-MCh
Slope, units 0.54 ⫾ 0.18 0.66 ⫾ 0.17
Intercept, l/s 1.76 ⫾ 0.58 1.18 ⫾ 0.39*
MP-HDM
Slope, units 0.79 ⫾ 0.08§ 0.53 ⫾ 0.11*
Intercept, l/s 1.16 ⫾ 0.66§ 1.15 ⫾ 0.32
FEV1, 1-s forced expiratory volume; FVC, forced vital capacity;
FEV1/FVC, ratio of FEV1 to FVC; MCh, methacholine; PD20, provoc-
ative dose causing a FEV1 decrease of 20% from control; AU, arbi-
trary units. Slopes and intercepts are from linear regressions of
maximal (M) vs. partial (P) flows recorded at all steps of MCh and
HDM challenges. Significant difference vs. rhinitis: * P ⬍ 0.05;
† P ⬍ 0.01; ‡ P ⬍ 0.001. Significant difference vs. MP-MCh, § P ⬍
0.05.
J Appl Physiol • VOL 91 • SEPTEMBER 2001 •
Fig. 1. Mean regression lines of maximal vs. partial flows during
challenges, calculated by solving the linear model y ⫽ a ⫹ bx, where
a is mean intercept and b is mean slope from Table 3. The extreme
x values for each line are the mean values of partial flow at control
and at challenge end point. Solid lines indicate methacholine chal-
lenge; dashed lines indicate house dust mite challenge. A, asthmatic
subjects; R, rhinitic subjects. See Table 3 for statistical differences.
Note the higher maximal flows for the same partial flows in R than
in A during MCh challenge, suggesting a greater bronchodilator
effect of deep inhalation. Note also the increase in slope in R during
allergen challenge, indicating a progressive reduction of the bron-
chodilator effect of deep inhalation with increasing allergen dose.
asthma when bronchoconstriction was induced by
MCh, as indicated by the significantly (P ⬍ 0.05)
higher MP intercept, but decreased progressively when
bronchoconstriction was induced by HDM, as indicated
by the significantly (P ⬍ 0.05) steeper MP slope.
DISCUSSION
The new findings of this study are that 1) the re-
sponse to MCh better separated asthmatic from rhi-
nitic subjects than did the response to HDM, 2) the
bronchodilator effect of DI was greater in rhinitis than
in asthma during MCh challenge, but it progressively
decreased in rhinitis during HDM challenge, and 3)
bronchial responsiveness to HDM was greater in
asthma than in rhinitis, even if baseline airway inflam-
mation and remodeling were similar. In addition, we
confirm and extend previous studies (17), showing that
the bronchi of rhinitic subjects developed an inflamma-
tory response to inhaled allergen that was similar to
that of asthmatic subjects.
Comments on methodology. This study has some
limitations. First, inflammatory cell data were ob-
tained only 24 h after the HDM inhalation. Therefore,
differences in the timing of inflammatory cell influx
into the airways cannot be evaluated. However, enu-
meration of blood eosinophils in seven subjects (4 with
asthma and 3 with rhinitis alone) showed a nonsignifi-
cantly different decrease 3 h after HDM inhalation
(57 ⫾ 9% in asthmatic subjects vs. 70 ⫾ 30% in rhinitic
www.jap.org

Downloaded from www.physiology.org/journal/jappl by ${individualUser.givenNames} ${individualUser.surname} (185.050.250.187) on December 26, 2018.

 BRONCHIAL RESPONSE TO ALLERGEN IN RHINITIS AND ASTHMA
subjects), suggesting a similar early recruitment of
eosinophils from blood in both groups. Second, inflam-
matory cells lying in the bronchial mucosa deeper than
100 ␮m below the basement membrane were not eval-
uated, and differences cannot be excluded. Third, no
relationship between inflammatory response and HDM
dose could be determined, as it would have been ethi-
cally unacceptable to have repeated HDM challenges
and bronchoscopies. Fourth, in this study, biopsy spec-
imens were taken at a single site and assumed to be
representative of airway remodeling throughout the
lung. This assumption seems to be justified by the
findings of Bradley et al. (3) and Jeffery et al. (11).
Finally, only subjects with mild asthma were studied,
and the findings cannot be extrapolated to more severe
disease.
Comments on results. In asthma, the degree of aller-
gic sensitization and the degree of bronchial respon-
siveness are independent determinants of both early
and late bronchoconstrictor responses to allergen (5).
In the present study, neither blood nor BAL IgE levels
were significantly different between asthmatic and rhi-
nitic subjects. For the same degree of systemic allergic
sensitization, as revealed by serum-specific IgE level
and/or skin test response, the bronchoconstrictor re-
sponse to allergen may be expected to be determined by
local factors, such as baseline bronchial inflammation,
allergen-induced bronchial inflammation, and mechan-
ical response.
In the present study, the degree of baseline bronchial
inflammation was not greater in asthma than in rhi-
nitis, but the dose of HDM required to cause a bron-
choconstrictor response (PD20) was lower in the former.
This suggests that the bronchoconstrictor response to
allergen is not critically dependent on the presence of
inflammatory cells or mediators in the airways before
challenge. The thickness of RBM was above the re-
ported normal values (19) but with no difference be-
tween groups, suggesting similar degrees of airway
wall remodeling (10). The functional consequences of
RMB thickening are complex and are discussed in a
companion paper (14).
As in a previous study (17), the inflammatory re-
sponse to allergen was similar in asthma and rhinitis,
but the HDM dose given to rhinitic subjects was
greater than that given to asthmatic subjects. Possible
reasons for a similar inflammatory response to differ-
ent doses of allergen in subjects with similar allergic
sensitization may be due to the compartmentalization
of the inflammatory-immune response with differences
in IgE receptor density on metachromatic cells, vascu-
lar leakage, or release of chemotactic substances from
resident cells, including smooth muscle cells. Whatever
the underlying mechanism, these data suggest, al-
though they do not prove, a different bronchial inflam-
matory responsiveness to allergen between asthma
and rhinitis.
An important, new finding of this study is that the
two groups differed more in respect to bronchial re-
sponsiveness to MCh than to HDM. All asthmatic
subjects were hyperresponsive to MCh and had a pos-
J Appl Physiol • VOL
Downloaded from www.physiology.org/journal/jappl by ${individualUser.givenNames} ${individualUser.surname} (185.050.250.187) on December 26, 2018.
1033
itive response to HDM. By contrast, the majority of
rhinitic subjects exhibited a bronchial responsiveness
to MCh within the normal range but a positive re-
sponse to HDM. It should be noted that falsely positive
responses in rhinitis are unlikely to have occurred
owing to the rather large cutoff value chosen, i.e., a
20% decrease of FEV1. MCh is a bronchoconstrictor
stimulus acting directly on airway smooth muscle. Al-
lergen is an indirect stimulus acting via the release of
a number of mediators, which may cause airway nar-
rowing by other mechanisms in addition to airway
smooth muscle contraction. A greater response to MCh
in asthma than in rhinitis is, therefore, suggestive of a
greater airway smooth muscle responsiveness. The ob-
served effects of DI on airway caliber are in line with
this interpretation. The magnitude of the bronchodila-
tor effect of DI is believed to be directly related to
airway hysteresivity and to the amplitude of stretching
exerted by lung parenchyma (7). A direct contractile
stimulus to airway smooth muscle should result in an
increase in hysteresivity only, thus fostering the bron-
chodilator effect of DI. An indirect bronchoconstrictor
stimulus may also cause airway wall edema, thus re-
ducing the magnitude of airway stretching and blunt-
ing the bronchodilator effect of DI. The greater MP
intercept during MCh challenge in rhinitis than in
asthma suggests that the ability to distend contracted
airway smooth muscle was greater in the former. Fur-
thermore, the similarity between the effects of DI dur-
ing MCh and HDM challenges in asthmatic subjects
suggests airway smooth muscle contraction as the ma-
jor determinant of response to HDM in this group. By
contrast, the increased MP slope during HDM in rhi-
nitis suggests a progressive reduction of the broncho-
dilator effect of DI with the increasing allergen dose.
This suggests a significant contribution of other mech-
anisms, such as airway wall edema and vascular leak-
age, possibly uncoupling airways from lung paren-
chyma (12). The tendency for a greater eosinophil
influx into rhinitic than asthmatic bronchi is consistent
with a somehow greater inflammatory response in the
former, likely related to the much greater dose of
allergen received. Another possibility is that the differ-
ence in the ability of DI to dilate constricted airways
between rhinitic and asthmatic subjects is due to a
relative difference in epithelial function, as suggested
by a tendency for less RBM surface being covered by
intact epithelium in the latter.
In conclusion, the results of this study suggest that
different mechanical responses, possibly related to air-
way smooth muscle responsiveness (18), may contrib-
ute to the different airway response to allergen, and
perhaps in symptoms, between asthma and rhinitis,
although different inflammatory responses cannot be
excluded.
The authors gratefully acknowledge Dr. Riccardo Pellegrino for
valuable comments on the manuscript.
This study was supported in part by grants from Ministero
dell’Università e della Ricerca Scientifica e Tecnologica (Rome, Italy)
(to V. Brusasco and G. W. Canonica) and by Associazione per la
Ricerca delle Malattie Immunologiche ed Allergiche (Genoa, Italy).
91 • SEPTEMBER 2001 • www.jap.org
 1034 BRONCHIAL RESPONSE TO ALLERGEN IN RHINITIS AND ASTHMA
REFERENCES
1. American Thoracic Society. Standards for the diagnosis and
care of patients with chronic obstructive pulmonary disease
(COPD) and asthma. Am Rev Respir Dis 136: 225–244, 1987.
2. Bonavia M, Crimi E, Quaglia A, and Brusasco V. Compari-
son of early and late asthmatic response between patients with
allergic rhinitis and mild asthma. Eur Respir J 9: 905–909, 1996.
3. Bradley BL, Azzawi M, Jacobson M, Assoufi B, Collins JV,
Irani A-M, Schwartz LB, Durham SR, Jeffery PK, and Kay
AB. Eosinophils, T-lymphocytes, mast cells, neutrophils, and
macrophages in bronchial biopsy specimens from atopic subjects
with asthma: comparison with biopsy specimens from atopic
subjects and relationship to bronchial hyperresponsiveness. J
Allergy Clin Immunol 88: 661–674, 1991.
4. Burney PGJ, Chinn S, Britton JR, Tattersfield AE, and
Papacosta AO. What symptoms predict the bronchial response
to histamine? Evaluation in a community survey of the Bron-
chial Symptoms Questionnaire (1984) of the International Union
Against Tubercolosis and Lung Disease. Int J Epidemiol 18:
165–173, 1989.
5. Crimi E, Brusasco V, Losurdo E, and Crimi P. Predictive
accuracy of late asthmatic reaction to dermatophagoides ptero-
nyssinus. J Allergy Clin Immunol 78: 908–913, 1986.
6. Fish JE, Rosenthal RR, Lichtenstein LM, and Norman PS.
Quantitative inhalation bronchial challenge in ragweed hay fe-
ver patients: a comparison with ragweed-allergic asthmatics. Am
Rev Respir Dis 113: 579–586, 1976.
7. Fredberg JJ, Inouye D, Miller B, Nathan M, Jafari S,
Raboudi SH, Butler JP, and Shore SA. Airway smooth mus-
cle, tidal stretches, and dynamically determined contractile
states. Am J Respir Crit Care Med 156: 1752–1759, 1997.
8. Giannarini L and Maggi E. Decrease of allergen-specific T-cell
response induced by local nasal immunotherapy. Clin Exp Al-
lergy 28: 404–412, 1998.
9. Grossman J. One airway one disease. Chest 111: 11–16, 1996.
10. James AL, Pearce-Pinto G, Elliot J, and Carroll N. The
relationship between basement membrane and airway wall di-
J Appl Physiol • VOL
Downloaded from www.physiology.org/journal/jappl by ${individualUser.givenNames} ${individualUser.surname} (185.050.250.187) on December 26, 2018.
mensions in asthma. Am J Respir Crit Care Med 161: A110,
2000.
11. Jeffery PK, Godfred RW, Ädelroth E, Nelson F, Rogers A,
and Johansson S-A. Effects of treatment on airway inflamma-
tion and thickening of basement membrane reticular collagen in
asthma. Am Rev Respir Dis 145: 890–899, 1992.
12. Macklem PT. A theoretical analysis of the effect of airway
smooth muscle load on airway narrowing. Am J Respir Crit Care
Med 153: 83–89, 1996.
13. Melillo G, Bonini S, Cocco G, Davies RJ, De Monchy JGR,
Frolund L, and Pelikan Z. EAACI provocation tests with
allergens. Allergy 52, Suppl 35: 1–35, 1997.
14. Milanese M, Crimi E, Scordamaglia A, Riccio A, Pellegrino
R, Canonica GW, and Brusasco V. On the functional conse-
quences of bronchial basement membrane thickening. J Appl
Physiol 91: 1035–1040, 2001.
15. Pellegrino R, Wilson O, Jenouri G, and Rodarte JR. Lung
mechanics during induced bronchoconstriction. J Appl Physiol
81: 964–975, 1996.
16. Quanjer PH, Tammelin GJ, Cotes JE, Pedersen OF, Peslin
R, and Yernault JC. Lung volumes and forced ventilatory
flows. Report Working Party “Standardization of Lung Function
Tests” European Coal and Steel Community. Eur Respir J 6,
Suppl 16: 5–40, 1993.
17. Shaver JR, O’Connor JJ, Pollice M, Cho SK, Kane GC,
Fish JE, and Peters SP. Pulmonary inflammation after seg-
mental ragweed challenge in allergic asthmatic and nonasth-
matic subjects. Am J Respir Crit Care Med 152: 1189–1197,
1995.
18. Solway J and Fredberg JJ. Perhaps airway smooth muscle
dysfunction contributes to asthmatic bronchial hyperresponsive-
ness after all. Am J Respir Cell Mol Biol 17: 144–146, 1997.
19. Vignola AM, Chanez P, Campbell AM, Souques F, Lebel B,
Enander I, and Bousquet J. Airway inflammation in mild
intermittent and in persistent asthma. Am J Respir Crit Care
Med 157: 403–409, 1998.
91 • SEPTEMBER 2001 • www.jap.org