J ALLERGY CLIN IMMUNOL

VOLUME 125, NUMBER 2

Abstracts AB91

Kinetics of the Immune Response in Childhood Eczema

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357 J. A. Wisniewski, T. Song, A. Reefer, M. Moloney, J. Patrie, S.

Satinover, D. Murphy, P. Heymann, J. Woodfolk; University of Virginia,

Charlottesville, VA.

RATIONALE: Eczema in childhood may be a harbinger of the ‘‘atopic

march.’’ The objective of this long-term study is to rigorously analyze

changes within the B cell and T cell compartments that could predict dis-

ease progression.

METHODS: Serum markers were measured in children with and without

eczema (ages 5 months to 18 years) and the phenotype of circulating T cells

analyzed by flow cytometry (ages 2 to 18 years).

RESULTS: Infants (<24 months (n516)) and children (>24 months

(n526)) with eczema exhibited elevated total IgE (gm580.5 IU/ml and

947 IU/ml* respectively) as compared to those without (40.1 IU/ml,

n525) (*p<0.001). Increased serum TARC levels and eosinophilia were

also evident from early infancy. Regression modeling revealed a rapid

rise in IgE antibody titers to milk and cat allergen starting before age

1 which plateaued by age 6. By contrast, IgE antibodies to dust mite, rye

and ragweed developed later and titers increased in a linear fashion through

late childhood with the most marked rise occurring for dust mite.

Regulatory T cells (CD127loFoxp3+) were scarce at age 3, but readily de-

tectable at age 6, regardless of disease status; however, while Foxp3+CD4+

T cells increased with age (r2 50.583, p50.029), there was a trend towards

decreased numbers in eczema (p50.07). In addition, increased expression

of CD25 on skin-homing (CLA+CD4+) T cells and increased CLA+ T cells

within the CD25hi subset was evident in children with eczema.

CONCLUSIONS: Differential IgE antibody kinetics associated with ec-

zema coincide with maturational and disease-specific changes in skin-

homing and regulatory T cell types.

358 B8 Improves Atopic Dermatitis-like Skin Lesion In Nc/NgaMice

H. Lee, J. Lee, H. Ha, D. Jung, M. Lee, N. Lee, J. Lee, C. Seo, H. Shin;

Korea Institute of Oriental Medicine, Deajeon, REPUBLIC OF KOREA.

RATIONALE: The topical steroids are commonly used in the treatment of

AD, but chronic use was discourage due to potential side effects such as

tachyphylaxis and symptomatic rebound following discontinuation of ther-

apy. We examined the effective activity of B8 in DfE-treated NC/Nga mice

so that want to suggest new safety candidate drug.

METHODS: The present study investigated whether the elevated expres-

sion level of plasma and chemokine genes are associated with AD using

DfE-treated NC/Nga mice. We examined the effective activity by histolog-

ical finding and TNF-a/IFN-g-induced keratinocyte (HaCaT) cell line.

RESULTS: Topical application of B8 significantly reduced the severity of

AD-like skin and ear lesion in DfE-treated NC/Nga mice compare with

positive control; protopic�. The B8 treatment significantly down-regulated

not only the plasma level of IgE and IL4 but also chemokines genes related

AD; TARC, MDC and RANTES in TNF-a/IFN-g-induced HaCaT.

CONCLUSIONS: Based on these results, it might be assumed that topical

application of B8 has systemic effect as well as local effects in the AD-like

lesions in NC/Nga mice. The topical application of B8 could improve the

AD-like lesions in NC/Nga mice by Th1 related cytokines. Taken together

it is suggested that topical application of B8 may be a novel approach to the

therapy of AD.

359 Development Of In Vitro T Cell Priming Assays ForIdentification Of Contact Allergens And RespiratorySensitizers

S. F. Martin1, P. R. Esser1, S. Schmucker2, D. Pennino3, R. Geiger4, E.

Maggi5, L. Dietz6, H. Thierse6, A. Richter2, A. Cavani3, F. Sallusto4; 1Uni-

versity Medical Center Freiburg, Freiburg, GERMANY, 2Miltenyi Biotec

GmbH, Bergisch-Gladbach, GERMANY, 3IDI-IRCCS, Rome, ITALY,4IRB, Bellinzona, SWITZERLAND, 5University of Florence, Florence,

ITALY, 6University Medical Center Mannheim, Mannheim, GERMANY.

RATIONALE: In vitro alternatives to replace animal testing in the identi-

fication of skin and respiratory sensitizers are urgently needed due to leg-

islation and for ethical reasons. One of the hallmarks of chemical contact

allergens and respiratory sensitizers is their ability to induce T cell re-

sponses. Our goal is to develop in vitro T cell priming assays within the

EU project Sens-it-iv that allow to identify such allergens.

METHODS: Autologous sorted naive T cells are primed with autologous

monocytes or dendritic cells. Contact or protein allergens are added to the

culture or DC are modified with contact allergens. Restimulation with an-

tigen is performed and T cell proliferation and cytokine production are

used to determine allergen specific T cell responses.

RESULTS: In vitro priming of naive human T cells with DC and contact

allergens or amplification of rare T cells to detect protein allergens results

in antigen-specific CD4+ and CD8+ T cell responses that allow in vitro

identification of chemical and protein allergens. Strengths and weaknesses

of the assays will be discussed.

CONCLUSIONS: In vitro T cell priming assays are of potential use as

second line tests within a tiered strategy for the identification of skin and

respiratory sensitizers, risk assessment and replacement of animal testing.

360 Correlation Of Serum Levels Of Th1 And Th2 Cytokines WithDisease Activity In Patients With Atopic Dermatitis

G. N. Drannik1, A. I. Kurchenko1, V. I. Omelchuk1, L. M. DuBuske2;1National Medical University, Kiev, UKRAINE, 2Immunology Research

Institute of New England, Gardner, MA.

RATIONALE: Atopic dermatitis (AD) involves both Th2 and Th1 im-

mune responses. This study assesses Th1 and Th2 serum cytokine levels

in relationship to disease severity.

METHODS: Fifty-five patients with AD were assessed. Clinical severity

scores of AD were monitored by using six-area, six-sign scores. Blood sera

were aliquotted, frozen at -808C, and thawed immediately prior to analysis.

Quantification of IL-18, IFN-g, IL-2, IL-12 and IL-10 (Immunotech, Co.,

Marseille, France) were performed by ELISA.

RESULTS: Serum IL-18, detectable in all AD patients and healthy con-

trols, was significantly greater in AD patients (252 pg/mL) versus controls

(106 pg/mL). Serum IL-18 levels also correlated well with the clinical re-

sponses in AD subjects. Serum IL-10, detectable in 45 of the 55 AD pa-

tients and in all 30 of the controls, did not differ between AD and

control subjects. Serum IL-12 was detected in all 55 AD patients (1.47

pg/mL) and in all 30 (0.5 pg/mL) of the controls. Serum IL-2 was detected

in 49 of the 55 AD patients (15.3 pg/mL) and in all of 30 controls (6.9 pg/

mL). There were no significant differences in IFN-g levels in AD versus

control subjects.

CONCLUSIONS: Sera IL-18 levels provided the most significant dis-

crimination between AD and control subjects and correlated with clinical

disease severity, suggesting that sera IL-18 levels may be useful to monitor

AD activity.