Developmental cytokine response profiles and the clinical and immunologic expression of atopy during the first year of life.

Abstract

Allergic diseases have been linked to abnormal patterns of immune development, and this has stimulated efforts to define the precise patterns of cytokine dysregulation that are associated with specific atopic phenotypes.
Cytokine-response profiles were prospectively analyzed over the first year of life and compared with the clinical and immunologic expressions of atopy.
Umbilical cord and 1-year PBMCs were obtained from 285 subjects from allergic families. PHA-stimulated cytokine-response profiles (IL-5, IL-10, IL-13, and IFN-gamma) were compared with blood eosinophil counts and total and specific IgE levels (dust mites, cat, egg, Alternaria species, peanut, milk, and dog) at age 1 year and at the development of atopic dermatitis and food allergy.
For the cohort as a whole, cytokine responses did not evolve according to a strict TH1 or TH2 polarization pattern. PHA-stimulated cord blood cells secreted low levels of IL-5 (2.1 pg/mL), moderate levels of IFN-gamma (57.4 pg/mL), and greater amounts of IL-13 (281.8 pg/mL). From birth to 1 year, IL-5 responses dramatically increased, whereas IL-13 and IFN-gamma responses significantly decreased. Reduced cord blood secretion of IL-10 and IFN-gamma was associated with subsequent sensitization to egg. In addition, there was evidence of TH2 polarization (increased IL-5 and IL-13 levels) associated with blood eosinophilia and increased total IgE levels by age 1 year.
These findings demonstrate that cytokine responses change markedly during the first year of life and provide further evidence of a close relationship between TH2 skewing of immune responses and the incidence of atopic manifestations in children.

Full text

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740
Mechanisms of allergy
Developmental cytokine response
profiles and the clinical and
immunologic expression of atopy during
the first year of life
William A. Neaville, MD,a,bChristopher Tisler, BS,aAbhik Bhattacharya, MS,d
Kelly Anklam, BS,aStephanie Gilbertson-White, BSN, RN,a
Rebekah Hamilton, MSN, RN,aKiva Adler, BS, RN,aDouglas F. DaSilva, BS,a
Kathy A. Roberg, MS, RN,aKirstin T. Carlson-Dakes, MEd, RN,a
Elizabeth Anderson, MA, BSN,aDebbie Yoshihara, BS,cRonald Gangnon, PhD,c
Lance D. Mikus, BS,bLouis A. Rosenthal, PhD,bJames E. Gern, MD,aand
Robert F. Lemanske, Jr, MDa,bMadison, Wis
Background: Allergic diseases have been linked to abnormal
patterns of immune development, and this has stimulated
efforts to define the precise patterns of cytokine dysregulation
that are associated with specific atopic phenotypes.
Objective: Cytokine-response profiles were prospectively ana-
lyzed over the first year of life and compared with the clinical
and immunologic expressions of atopy.
Methods: Umbilical cord and 1-year PBMCs were obtained
from 285 subjects from allergic families. PHA-stimulated
cytokine-response profiles (IL-5, IL-10, IL-13, and IFN-γ) were
compared with blood eosinophil counts and total and specific
IgE levels (dust mites, cat, egg, Alternaria species, peanut,
milk, and dog) at age 1 year and at the development of atopic
dermatitis and food allergy.
Results: For the cohort as a whole, cytokine responses did not
evolve according to a strict TH1 or TH2 polarization pattern.
PHA-stimulated cord blood cells secreted low levels of IL-5
(2.1 pg/mL), moderate levels of IFN-γ (57.4 pg/mL), and
greater amounts of IL-13 (281.8 pg/mL). From birth to 1 year,
IL-5 responses dramatically increased, whereas IL-13 and
IFN-γ responses significantly decreased. Reduced cord blood
secretion of IL-10 and IFN-γ was associated with subsequent
sensitization to egg. In addition, there was evidence of TH2
polarization (increased IL-5 and IL-13 levels) associated with
blood eosinophilia and increased total IgE levels by age 1 year.
Conclusion: These findings demonstrate that cytokine respons-
es change markedly during the first year of life and provide
further evidence of a close relationship between TH2 skewing
of immune responses and the incidence of atopic manifesta-
tions in children. (J Allergy Clin Immunol 2003;112:740-6.)
Key words: Cytokines, TH2, immune development, allergy, IgE
Recent observations have stimulated research efforts to
further define the relative importance and pathophysio-
logic contributions of cytokine dysregulation (so-called
TH1/TH2 imbalance) to the development of various atopic
phenotypes.1Although questions remain regarding the
full effect of a TH1/TH2 dysregulation in established
atopic diseases, the contribution of cytokine polarization
to the inception and evolution of various atopic diseases,
including asthma, has received more uniform support. For
example, there is evidence to suggest that at birth, possi-
bly because of placentally derived TH2 trophic factors,
allergen-induced mononuclear cell (MNC) responses are
skewed toward a TH2-like phenotype,2and IFN-γ
responses are particularly low.3,4Furthermore, IFN-γ
responses gradually increase in magnitude in normal
infants, whereas in allergic infants IFN-γ responses are
reduced at birth, and their development is delayed.2,5,6As
a result, this reduced IFN-γ generation causes a TH1/TH2
imbalance that might be a risk factor for the subsequent
development of allergic disease, asthma, or both.2,3,7,8
Although the concept of a TH1/TH2 imbalance is an
attractive theory, cord blood IL-13 responses also appear
to be suppressed in children who go on to have atopic dis-
ease.9Collectively, these data suggest a complex interplay
of TH1/TH2 responses in relationship to immune matura-
tion and the development of allergic disease.
Although these observations are of considerable inter-
est, it is important to note that they were made in a small
number of patients who displayed a variety of different
atopic phenotypes. To extend these observations, the pre-
sent study, Childhood Origins of Asthma, is designed to
analyze the development of cytokine-response profiles
beginning at birth in a large cohort of high-risk children
and to determine their relationships with clinical and bio-
logic markers of atopy and eventually asthma. To test the
hypothesis that different atopic phenotypes will be asso-
From the Departments of aPediatrics,bMedicine, and cBiostatistics and Med-
ical Informatics and dthe General Clinical Research Center, University of
Wisconsin-Madison.
Supported by National Institutes of Health grants #1R01HL61879-01 and
#1P01HL70831-01 and General Clinical Research Center grant National
Institutes of Health/National Center for Research Resources 2 MOI RR
03186-16. Additional financial support was received from the following:
the Wisconsin Medical School, the University of Wisconsin Medical School
Department of Pediatrics, the University of Wisconsin Graduate School,
AstraZeneca, Aventis, GlaxoSmithKline, Medimmune, and Merck.
Received for publication May 13, 2003; revised June 17, 2003; accepted for
publication June 19, 2003.
Reprint requests: William Neaville, MD, K4/916 University of Wisconsin
Hospital, 600 Highland Ave, Madison, WI 53792-9988.
© 2003 American Academy of Allergy, Asthma and Immunology
0091-6749/2003 $30.00 + 0
doi:10.1067/mai.2003.1716

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VOLUME 112, NUMBER 4
Neaville et al 741
Mechanisms of allergy
ciated with distinct TH2 patterns of cytokine develop-
ment, subjects were prospectively evaluated for the first
year of life with historical questionnaires, physical exam-
inations, and laboratory studies to compare cytokine
responses with the development of total and allergen-
specific IgE, blood eosinophilia, atopic dermatitis (AD),
and food allergy.
METHODS
Experimental design
Study subjects included 285 children with at least one parent
with respiratory allergies (defined as ≥1 positive aeroallergen skin
test responses), asthma (defined historically), or both (Table I).
After obtaining informed consent, subjects were enrolled at birth
(from November 1998 through March 2000) and followed prospec-
tively for at least 1 year. Data collected included parent and child
questionnaires regarding health histories, with a specific focus on
atopic diseases, behaviors affecting health, and environmental expo-
sures. Physical examinations were performed at regularly scheduled
times (generally 2, 4, 6, 9, and 12 months) by the subjects’primary
physicians, and these records were evaluated to document the devel-
opment of AD and food allergy. Blood samples were collected at
birth (cord blood) and at age 1 year. Laboratory studies included
cytokine-response profiles from stimulated cord blood MNCs and
PBMCs at age 1 year. Additional studies performed at age 1 year
included RAST testing, total IgE level measurement, and peripher-
al blood eosinophil count measurement. This study was approved
by the University of Wisconsin Human Subjects Committee.
Collection of blood samples
Cord blood samples were collected on each of the study partici-
pants. After the birth of the child, the umbilical cord was ligated
before delivery of the placenta. The cord blood was then collected
into sterile heparinized tubes that were labeled with the subject
name and time of collection and kept at room temperature. Cord
blood specimens were processed within 16 hours of collection.
Peripheral blood samples (2.5-10 mL) were collected at age 1 year
by means of venipuncture at the antecubital site of the arm. Lido-
caine 2.5% and prilocaine 2.5% (EMLA, AstraZeneca) was applied
to the area 45 to 60 minutes before phlebotomy. The blood was col-
lected in sterile heparinized tubes, kept at room temperature, and
then processed on the day of collection.
MNC stimulation
MNCs were separated by means of density centrifugation (LSM
Lymphocyte Separation Medium; ICN Biomedicals Inc, Aurora,
Ohio), and plasma was removed and stored at –80°C in labeled
microcentrifuge tubes. The MNCs were suspended (106cells/mL) in
RPMI-1640 supplemented with 10% FBS (Hyclone), L-glutamine (2
mmol/L), penicillin (100 U/mL), and streptomycin (100 µg/mL) and
incubated (24 hours at 37°C and 5% CO2) in a 24-well, flat-bottom,
cell-culture plate (1 mL/well; Corning Inc, Corning, NY) with PHA
(5 µg/mL; Sigma, St Louis, Mo) or medium alone. The PHA was
placed in aliquots in single-use portions and frozen at –80°C at the
beginning of the study so that the same stimulant was used through-
out the study. After the culture supernatants were collected, they
were frozen (–80°C) in aliquots pending analysis for cytokines.
Cytokine ELISA
Levels of IFN-γ, IL-5, IL-10, and IL-13 in culture supernatants
were evaluated by means of ELISA (Pharmingen, San Diego,
Calif). The manufacturer’s protocol was followed except that the
sample volume was reduced to 50 µL. The sensitivities of the
ELISAs are as follows: IFN-γ, 3.1 pg/mL; IL-5, 1.9 pg/mL; IL-10,
7.8 pg/mL; and IL-13, 3.1 pg/mL. Duplicate wells were run for each
sample, and mean values are reported.
Total and allergen-specific IgE
Fluoroenzyme immunoassays (FEIAs) were performed on plas-
ma obtained from 1-year peripheral blood samples by using an
automated instrument (Unicap 100; Pharmacia and Upjohn Diag-
nostics, Kalamazoo, Mich) to determine total and specific IgE lev-
els. Specific IgE levels were determined for 2 species of dust mite
(Dermatophagoides farinae and Dermatophagoides pteronyssinus),
cat dander, egg white, Alternaria alternata, peanut, milk, and dog.
The sensitivity for detection of specific IgE was 0.35 kU/L, and val-
ues greater than or equal to 0.35 kU/L were considered positive. The
sensitivity for detection of total IgE was 2 kU/L.
Measurement of blood eosinophils
Total white blood cell counts were performed by using an auto-
mated instrument. Blood smears from peripheral blood samples
were stained (Wright-Giemsa stain), and a 200-count white blood
cell differential was performed. The absolute eosinophil counts
were then calculated for all peripheral blood samples.
AD and food allergy definitions
AD was defined as physician diagnosed either by means of doc-
umentation by a health care provider on the medical record or by
means of parental report of physician-diagnosed AD on the histori-
cal questionnaires. Three groups were defined and used for this
study: (1) children with no AD anytime within the first year of life;
(2) children with AD documented at any point within the first year;
and (3) children with active disease at age 1 year.
Food allergy was defined by using allergen-specific IgE test
results (FEIA) and historical reports (parental reports, physician
documentation, or both). Three groups were defined: (1) probable
food allergy includes a positive FEIA and a convincing history,
defined as a reproducible adverse response that is temporally relat-
ed to a specific food and involves typical organ systems10; (2) ques-
tionable food allergy includes a positive FEIA result with an inde-
terminate history or a negative FEIA result with a suggestive histo-
ry; and (3) no food allergy includes a negative FEIA result with a
negative history or a positive FEIA result with no adverse reactions
to the food or foods in question.
Statistical analysis
Continuous parameters are summarized by using medians and
interquartile ranges. Dichotomous parameters are summarized as per-
centages. Changes in the cytokine-response profile from cord blood
to age 1 year were assessed by using the Wilcoxon signed-rank test.
Associations between cytokine-response profiles (cord blood, age 1
year, or change from cord blood to age 1 year) and biologic markers
of atopy (absolute eosinophil counts or total IgE levels) were assessed
by using the Spearman rank correlation coefficient. Rates of allergic
sensitization (≥1 positive FEIA result) were calculated for quartiles of
cord blood cytokines. Differences in cytokine-response profiles on
the basis of the presence or absence of allergic sensitization at age 1
year were assessed by using the Wilcoxon rank sum test. Similar
analyses were conducted for outcomes of AD during the first year of
life and active AD at the 1-year visit.
Abbreviations used
AD: Atopic dermatitis
FEIA: Fluoroenzyme immunoassay
MNC: Mononuclear cell

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742 Neaville et al
J ALLERGY CLIN IMMUNOL
OCTOBER 2003
Mechanisms of allergy
RESULTS
Immunologic development in the first year
of life
To begin to evaluate the immunologic development of
this high-risk cohort, we compared the cytokine secretion
patterns at birth (cord blood) with those obtained at age
1 year. Although the pattern of cytokine responses
changed significantly, the changes did not follow either a
TH1 or TH2 polarization pattern (Fig 1). PHA-stimulated
cord blood cells secreted low levels of IL-5 (median, 2.1
pg/mL), moderate levels of IFN-γ (57.4 pg/mL) and IL-
10 (100 pg/mL), and greater amounts of IL-13 (281.8
pg/mL). PHA-induced IL-5 responses markedly
increased from birth to 1 year (Fig 1, A), whereas IL-13
and IFN-γ responses decreased by 16% and 54%, respec-
tively (Fig 1, B and C). There was no significant change
in IL-10 responses in the first year of life (Fig 1, D).
Cytokine responses and biologic markers of
atopy
To compare cytokine-response profiles to biologic
markers of atopy at age 1 year, we first evaluated rela-
tionships between peripheral blood absolute eosinophil
counts and total IgE levels. Cord blood cytokine respons-
es did not correlate with either of these atopic markers at
age 1 year (Table II). In contrast, there were weak but
FIG 1. Interval changes for cytokine-response profiles from birth to 1 year for the entire cohort. The box rep-
resents the 25th and 75th percentiles, with medians indicated by horizontal lines, and the 5th and 95th per-
centiles are represented by filled circles.
TABLE I. Subject demographics
Maternal characteristics
Age (y)
History
Allergy
Asthma
Both
Ethnicity, white
Paternal characteristics
Age (y)
History
Allergy
Asthma
Both
Ethnicity, white
Child characteristics
Sex, male
Ethnicity, white
Birth weight (kg)
Head circumference (cm)
Apgar score
10
9
8
7
31.3 ± 4.8
43.7%
3.9%
36.2%
86.8%
32.9 ± 5.3
48.6%
2.1%
24.6%
86.8%
56.3%
86.8%
3.54 ± 0.54
34.6 ± 2.4
7.4%
80.9%
9.6%
2.1%
Values are presented as means ± SD for continuous variables and percent-
ages for categoric variables.

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VOLUME 112, NUMBER 4
Neaville et al 743
Mechanisms of allergy
significant positive correlations between 1-year IL-5 and
IL-13 responses and both markers of atopy and between
1-year IL-10 responses and the absolute eosinophil count
(Table II). Positive correlations were also noted between
changes in IL-5 and IL-13 responses from birth to 1 year
and both markers of atopy (Table II). In contrast, there
was no correlation between IFN-γ responses and either
eosinophilia or total IgE.
Cord blood cytokine responses were grouped accord-
ing to quartiles to determine whether cytokine responses
at birth predicted the risk of allergen sensitization at age
1 year. Infants with high IL-10 responses at birth were
significantly less likely to be sensitized to egg at age 1
year (Table III). There were no significant associations
between egg sensitization and 1-year cytokine responses.
Allergic sensitization in general, defined as detectable
IgE for any one of the 8 allergens tested, was not associ-
ated with specific patterns of cytokine responses (data
not shown).
Cytokine responses, AD, and food allergy
Cytokine-response profiles were evaluated in children
according to the presence or absence of AD or food aller-
gy. AD was associated with increased total IgE (17 vs 11
IU/mL, P = .02), egg-specific IgE (29% vs 8%, P <
.001), and a trend toward increased peripheral blood
eosinophils (237 vs 194 cells/mL, P = .07); however,
there were no significant associations between the pres-
ence of AD and cytokine-response profiles at birth or age
1 year (Table IV). Likewise, food allergy was not associ-
ated with significant alterations in polyclonal cytokine-
response profiles (Table IV).
DISCUSSION
The increase in prevalence of allergic airway diseases
over the last several years11-13has generated interest in
gaining a better understanding of the pathogenesis of
these disorders in infancy. Our findings extend the results
of previous studies and provide new information regard-
ing the development of polyclonal cytokine responses
relative to the incidence of clinical and immunologic
expression of atopy. For the group as a whole, polyclon-
al cytokine responses over the first year of life did not
evolve according to a strict TH1 or TH2 polarization pat-
tern. For example, there were opposite directional
changes in IL-5 (increase) versus IL-13 (decrease). In
contrast to the whole group, there was a TH2 polarization
in infants who had biologic markers of atopy, such as
eosinophilia (increased IL-5, IL-10, and IL-13) and
increased total IgE levels (increased IL-5 and IL-13).
Furthermore, reduced secretion of IL-10 by mitogen-
stimulated cord blood cells was identified as a risk factor
for sensitization to egg, which was the most common
sensitization noted in our cohort. Together, these findings
provide further evidence of a close relationship between
immune development (TH2 skewing) and the incidence
of atopic manifestations in children.
When evaluating the immune development of the
entire cohort, the most dramatic change noted was the
paucity of IL-5 secretion from cord blood cells and the
marked increase by age 1 year. The relatively high IL-13
responses from the stimulated cord blood MNCs indicate
that not all TH2-like cytokines are suppressed at birth.
Instead, this pattern suggests that IL-5 production might
be selectively inhibited at birth and then developmentally
cued by an as-yet-undefined postneonatal signal.
Other groups have also evaluated the development of
cytokine responses in infancy. Prescott et al2demonstrat-
ed that allergen-specific cytokine responses at birth
might be skewed toward a TH2 phenotype. Specifically,
TABLE II. Spearman rank correlation coefficients
between cytokine responses and biomarkers
Absolute eosinophil countTotal IgE
Cord blood
IFN-γ
IL-5
IL-10
IL-13
1 y
IFN-γ
IL-5
IL-10
IL-13
Change (1 y, cord blood)
IFN-γ
IL-5
IL-10
IL-13
0.079
0.006
–0.008
–0.017
0.108
–0.038
0.013
–0.033
–0.04
0.265‡
0.168*
0.264‡
–0.043
0.163†
0.08
0.16†
–0.115
0.26†
0.109
0.215†
–0.116
0.162†
0.074
0.165†
*P < .05.
†P < .01.
‡P < .001.
TABLE III. Cord blood cytokine responses and sensitiza-
tion to egg
Cytokine responses
(quartiles, pg/mL)
Sensitization to egg
(% positive)nP value
IFN-γ
<31.0
31.0-57.3
57.4-121.6
≥121.7
IL-5
<1.9
1.9-2.0
2.1-6.7
≥6.8
IL-10
<58.4
58.4-99.4
99.4-164.3
≥164.4
IL-13
<152.9
152.9-281.7
281.7-487.3
≥487.4
.079
71
71
72
70
25.7%
14.3%
11.3%
16.2%
.114
13720.1%
14.3%
11.8%
15.7%
7
69
71
.013
71
70
71
70
24.3%
17.6%
18.8%
7.1%
.293
71
71
71
70
21.4%
15.7%
14.5%
15.9%

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744 Neaville et al
J ALLERGY CLIN IMMUNOL
OCTOBER 2003
Mechanisms of allergy
allergen stimulation of cord blood cells upregulated IL-4,
IL-5, and IL-9 but not IFN-γ mRNA. In addition, aller-
gen-stimulated cord blood cells secreted high levels of
IL-10 (approximately 3000 pg/mL) and secreted low lev-
els (10 pg/mL) of IL-13. Additional studies by this group
indicated that, during development, there is a decrease in
TH2 responses to inhalant allergens in nonatopic chil-
dren, whereas there is an increased or persistent TH2
response in atopic children.14,15
IL-13 is produced from TH2 lymphocytes and plays a
role in inhibiting pro-inflammatory cytokine production,
in the induction of IgE isotype switching, and in vascular
cell adhesion molecule 1 expression.16-18Paradoxically,
studies have found that mitogen-stimulated7and aller-
gen-stimulated14,15cord blood MNC production of IL-13
is reduced in atopic children. In infancy there appear to
be age-dependent increases in allergen-specific IL-13
responses for the atopic children, whereas the responses
wane in nonatopic children.14,15Data from the present
study extend these findings by demonstrating significant
correlations between the development of TH2 skewing
(increased polyclonal IL-13 and IL-5 responses) and spe-
cific biologic markers of atopy: total IgE and peripheral
blood eosinophilia.
Although no statistically significant differences were
noted between cord blood cytokine responses and aller-
gen-specific IgE results grouped as a whole, reduced IL-
10 responses in cord blood were associated with an
increased frequency of egg allergen sensitization. In the
first year of life, sensitization is most frequently seen
with egg allergen, and this is a risk factor for the subse-
quent development of clinically apparent allergic dis-
eases, such as AD, allergic rhinitis, and asthma.19-22
Interestingly, IL-10 has been implicated in the develop-
ment of tolerance through several potential mechanisms,
including shifting antibody responses from IgE to
IgG423,24and promoting the development of T regulato-
ry cells.25,26
Although patterns of cytokine responses were clearly
associated with various immunologic aspects of atopy,
we did not find a significant association with AD or food
allergy. Previous studies of immune development in AD
have yielded conflicting results: some studies have
reported that AD is associated with reduced IFN-γ lev-
els,3,27whereas Halonen and Martinez8found no such
association. The divergent results of these studies are
likely due to differences in the subject population, how
AD was defined, and the length of follow-up. It is possi-
ble that a stronger relationship between AD and IFN-γ
production exists with more severe disease: many of the
children in our cohort had relatively mild disease com-
pared with subjects in other studies.3Also, the design of
the present study is a large prospective cohort, which
inherently requires the clinical assessment of several
investigators and historical data provided from the par-
ents to make the diagnosis of AD. This differs from other
studies in which the cohorts were smaller and a single
investigator could make the diagnosis of AD on the basis
of clinical findings.27We are continuing to monitor these
relationships because the clinical expression of these
atopic phenotypes appears de novo, regresses with time,
or both. In addition, we will be evaluating these out-
comes in relationship to the emergence of other atopic
phenotypes (eg, asthma and rhinitis) as well.
There are several important differences between our
study and previously published work. First, the cytokine-
response profiles for this study were evaluated after mito-
gen stimulation rather than specific allergen stimulation.
This enabled the measurement of cytokine protein secre-
tion rather than relying on the measurement of mRNA,
which has been reported for similar evaluations with
allergen-evoked responses.2,14,15Because the use of
allergen-specific stimulation would likely result in a sig-
nificant proportion of the cohort having no response at
birth, the polyclonal T-cell mitogen PHA was used to
maximize our ability to investigate the developmental
aspect of cytokine-response profiles beginning with cord
blood in the entire cohort.
Second, this study is relatively unique in that it is a
prospective evaluation of a large cohort of children who
all have a parental history of allergy or asthma. Although
this design could be criticized for the lack of inclusion of
a low-risk (by parental history) control group, more than
half of the study subjects have exhibited no biologic or
clinical evidence of atopy. We are intrigued by the fact
that the cytokine responses of the infants who are
TABLE IV. Cord blood cytokine responses compared with AD and food allergy
Atopic dermatitisFood allergy
Never
(n = 162)
Ever
(n = 123)
P value
(E vs N)
NoYesP value
(N vs Y)(n = 251)(n = 15)
Cord blood
IFN-γ
IL-5
IL-10
IL-13
Change (1 year, cord blood)
IFN-γ
IL-5
IL-10
IL-13
55.1 (32.9, 117.7)
2.2 (1.5, 6.5)
106.7 (66.5, 172.6)
290.7 (170.2, 490.8)
60 (30.7, 127.1)
1.9 (1.5, 7.1)
97.8 (49.5, 161.7)
280.4 (145.6, 457.0)
.93
.8
.43
.37
61.3 (33.8, 119.4)
2.1 (1.5, 6.3)
103.7 (59.6, 171.0)
281.8 (152.9, 486.0)
37.7 (14.7, 93.6)
1.5 (1.5, 7.8)
96.1 (35.1, 126.7)
316.9 (102.7, 511.9)
.12
.73
.3
.91
–27.1 (–86.2, 2.1)
124.7 (62.1, 237.5)
–16.5 (–75.8, 56.9)
–73.3 (–260.3, 116.0)
–34.8 (–94.1, –1.7)
156.1 (69.4, 244.8)
–5.2 (–51.7, 41.5)
–6.7 (–219.9, 149.9)
.34
.39
.55
.16
–32.4 (–92.3, 0.0)
137.2 (69.8, 247.3)
–5.8 (–67.4, 49.4)
–37.8 (–237.9, 122.6) –55.9 (–319.8, 77.1)
–15.7 (–52.7, 15.2)
110.3 (52.3, 179.3)
–24.3 (–60.7, 42.1)
.36
.15
.85
.49
Values represent median (range).

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VOLUME 112, NUMBER 4
Neaville et al 745
Mechanisms of allergy
expressing biologic markers of atopy (increased total IgE
and absolute eosinophil counts) by the first year of life
diverge from those of the children who are not and that
cytokine-response profiles when evaluated during infan-
cy are more likely to reflect the biologic rather than the
clinical expression of atopy. As such, we would predict
the continued study of this high-risk cohort will provide
a unique opportunity to comprehensively evaluate gene-
gene and gene-environment interactions that might facil-
itate not only the clinical expression of atopic phenotypes
but also the downregulation of these traits.
In summary, the results of our study demonstrate the
complex interplay of cytokine responses during infancy
in relationship to age (development), biologic markers of
atopy, and disease expression. Defining the immunolog-
ic characteristics of atopy in infancy and early childhood
continues to be an important research goal. By compar-
ing and contrasting the immunologic development for the
different atopic phenotypes in infancy, it might be possi-
ble to identify early immunologic characteristics, clinical
characteristics, or both that predispose to the develop-
ment of allergic diseases, such as asthma.
This project would not have been possible without the support
and cooperation of the following health care professionals: Gail
Allen, Conrad Andringa, Candye Andrus, Richard Anstett, Maribeth
Baker, Robert Baker, Adam Balin, Ann Behrmann, Patricia Bellis-
simo, Arnold Benardette, George Benton, Tom Best, Gregory Bills,
John Bohn, Connie Brandt, Don Breckbill, Don Buckstein, Rebec-
ca Bull, Renee Burk, Deirdre Burns, Robert Cape, Colleen Calvy,
Michael Cardwell, Susan Carson, Cally Christiansen, R. Christ-
mann, Timothy Chybowski, Marcus Cohen, Robert Cole, Alison
Craig, Alison Dahlrymple, James Davis, Nancy Deaton, Patricia
Deffner-Valley, Jean Demopolous, Gregory DeMuri, Kathleen
DeSantes, Sherri DeVries, Klaus Diem, Karla Dickmeyer, Sabine
Droste, Bruce Drummond, Paul Dvorak, Rebecca Eckland, M.
Bruce Edmonson, Anne Eglash, Susan Ehrlich, Lawrence Elfman,
Marguerite Elliott, Richard Ellis, Dolores Emspak, Sue Engel-
baugh, Aida Evans, Gordon Faulkner, Christopher Federman, Ellen
Flannery, Joseph Fok, Nadine French, Carolyn Fruehling, Ann
Ganch, Claire Gervais, Michelle Gigot, Jenny Hackforth-Jones,
Nancy Hagan, David Hann, Mark Hansen, Thomas Hartjes, Jean
Haughwout, William Heifner, Russell Hermus, Russell Hess, Karen
Hillery, Sherry Holtzmann, Barbara Hostetler, Catherine James,
Brenda Jenkin, Sandy Kamnetz, Peter Karofsky, Jennifer Kaufman,
Helen Kay, Holly Keevil, Jeffrey Keil, Catherine Kelley, Jeanine
Kies, Steve Kincaid, Gretchen Kind, Kristen Knoepke, J. Brendt
Kooistra, Paul Kornaus, Steven Koslov, Jeff Krawcek, Dean Kresge,
Ann Krigbaum, Robert Kriz, Karen Kronman, Gordon Kronquist,
Randy Krszjzaniek, Greg Landry, Mary Landry, Martha Lauster,
Karen Lentfer, Joanne Leovy, Stephen Lo, David Lonsdorf, Allan
Luskin, Francois Luyet, Joseph Mahoney, Dimitria Manesis, Daniel
Marley, Edward McCabe, Gwen McIntosh, Anne Means, Thomas
Meier, Frederic Melius, Theresa Mendoza, J. Michael, Eileen
Michaels, Bernard Micke, Kaye Mickelson, James Milford, Angie
Miller, Julie Mokhtar, Jonathan Morey, Vickie Mulkerin, Maureen
Murphy-Greenwood, Thomas Murwin, Eric Nagle, Elizabeth
Neary, Paul Neary, James Nettum, Susan Nondahl, Carolyn Ogland,
David Okada, Mark Olinger, Reid Olson, Kathleen Oriel, Sandra
Osborn, John Ouellette, Meg Parker, Daniel Paulson, Karen Pletta,
Amy Plumb, Beth Potter, Peter Pryde, William Ranum, Richard
Rice, David Ringdahl, Michael Ritter, Richard Roberts, Everett
Roley, Meriel Ronstadt, Karl Rudat, Sherwin Rudman, Jerry Ryan,
Julie Saxton, William Scheibel, Ben Schmidt, Rosemarie Schu-
macher, Julie Schurr, Gerald Shay, Debra Shenoi, James Shrop-
shire, Charles Shutt, Elma Sia, Linda Siewart, Theresa Sizer, Susan
Skochelak, Jeffrey Sleeth, Greg Smith, Sheryl Spitzer-Resnick,
Patricia Staats, Jennifer Stevens, Katherine Stewart, Mary Stoffel,
Joanne Taylor, Jonathan Temte, Stephen Thomas, Mark Timmer-
man, Ordean Torstenson, Mary-Anne Urtes, Thomas Varley,
Eleanor Vita, Lisa Wacholz, David Weber, Bonny Whalen, Margaret
Wilcots, Gary Williams, W. Michael Wilson, Robin Wright,
Michael Yaffe, and Kok-Peng Yu. The support and participation of
the following hospitals and clinics has been key to the success of the
project: the obstetric nursing staff at Meriter Hospital, St Mary’s
Medical Center, Fort Atkinson Memorial Health Services, Inc, St
Clare Hospital, Reedsburg Area Medical Center, and Sauk Prairie
Memorial Hospital. Additionally, the clinic staff from Physicians
Plus, Associated Physicians, Dean Medical Center, Group Health
Cooperative, and other clinics in southwestern Wisconsin seeing
individual Childhood Origins of Asthma families are appreciated.
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