Abstracts AB223

J ALLERGY CLIN IMMUNOL

VOLUME 135, NUMBER 2

Nasal Allergen Challenge (NAC) Induced Eosinophilia - the

Allergic Rhinitis Clinical Investigator Collaborative (AR-CIC)
Jenny Thiele, MSc1,2, Daniel Adams, BSc1, Mena Soliman, MBChB1,2,
Lisa Steacy, BSc1, Anne Ellis, MD, MSc, FAAAAI1,2; 1Allergy Research
Unit, Kingston General Hospital, Kingston, ON, Canada, 2Departments of
Medicine and Biomedical & Molecular Science, Queens University,
Kingston, ON, Canada.
RATIONALE: The Allergic Rhinitis Clinical Investigator Collaborative
(AR-CIC) is a Canadian initiative with the goal of performing standardized
nasal allergen challenge (NAC) to study the effects of therapeutic agents
for allergic rhinitis (AR), while allowing the identification of mechanisms
and biomarkers of AR. Various NAC protocols have been described
previously. The multiple cumulative allergen concentration (MCAC) NAC
protocol was shown to produce more robust symptom scores than a
cumulative allergen concentration (CAC) protocol. Here we examined
NAC-induced eosinophilia for these two protocols.
METHODS: 17 atopic and 12 non-atopic participants were enrolled for
this study. Atopic individuals presented with AR symptoms in ragweed
season and a supportive skin test response. During screening incremental
concentrations of ragweed allergen were administered until each participant reached the qualifying symptom cut-off. For the subsequent NAC one
week later, ten atopics were challenged with one dose of allergen
equivalent to the cumulative amount of allergen each received during
screening (CAC). Seven atopics received the cumulative of all preceding
allergen doses to the qualifying concentration (QC), followed by the QC 15
minutes later (MCAC). Non-atopics were challenged with a 1:2 ragweed
concentration. Nasal lavage samples were collected at baseline, 1 hour
(1H) and 6 hours (6H) post NAC to determine differential counts.
RESULTS: The eosinophil fraction was significantly increased in atopics
following NAC when compared to non-allergics at both 1H and 6H for the
CAC-protocol but only at 6H for the MCAC-protocol.
CONCLUSIONS: Even though the MCAC protocol establishes more
robust symptom scores, the CAC protocol appears to produce more
pronounced eosinophilia.

722

Differential Exposure of Eosinophils to Cytokines Leads to an

Activated Phenotype in the Airways Inducing Pulmonary IL-13
Responses That Are Eosinophil Dependent
Elizabeth A. Jacobsen, PhD, Dana C. Colbert, Katie R. Zellner, Cheryl
A. Protheroe, William E. LeSuer, Nancy A. Lee, PhD, James J. Lee, PhD;
Mayo Clinic Arizona, Scottsdale, AZ.
RATIONALE: Allergic respiratory inflammation is characterized by
immune responses and induced pathologies linked with increased airway
IL-13. Mice deficient (CCR3-/-, IL-5-/-) or devoid (PHIL, DdblGATA-1,
IL-5-/-eotaxin-1-/-) of eosinophils display reduced pulmonary IL-13 in
various mouse models of allergic respiratory inflammation. Moreover, previous studies have demonstrated that eosinophils are capable of both expressing IL-13 and promoting its expression by other resident lung cells.
METHODS: Purified blood-derived wild type or cytokine deficient (IL4-/- and IL-13-/-) eosinophils were either untreated or cytokine pre-treated
prior to adoptive transfer into the lungs of allergen challenged mice deficient (IL-5-/-) or devoid (PHIL) of eosinophils. Allergen-induced pulmonary changes in Th2 cytokine levels, histopathologies, and accumulation of
leukocyte populations were assessed.
RESULTS: Eosinophils with activation-state unique activities were
achieved by differential cytokine pre-treatment prior to adoptive transfer
into the lungs of allergen-treated IL-5-/- or PHIL mice. GM-CSF treated
eosinophils activated T cells and DCs in the lung draining lymph nodes,
yet did not induce either T cell recruitment to the lung or evidence of pulmonary inflammation. In contrast, eosinophils pre-treated with both IL-33
and GM-CSF were sufficient to restore pulmonary T cell accumulation as
well as induced allergen-mediated pathologies. We demonstrated in both
IL-5-/- and PHILmice that the Th2 pulmonary immune responses and
induced pathologies mediated by differentially activated eosinophils

were specifically dependent on the expression of IL-13 (but not IL-4) by

eosinophils.
CONCLUSIONS: Our data demonstrates an underappreciated and
significant role for eosinophil-derived IL-13 in inducing Th2 pulmonary
pathologies in mouse models of allergic respiratory inflammation.

723

Preferential Production of TNFa, Compared to IL-4 and IFNg,

By Differentiating Eosinophil-Basophil Cord Blood
Progenitors
Pia Reece, PhD1, Claudia C. K. Hui, PhD2, Judah A. Denburg, MD,
FRCPC, FAAAAI1; 1Division of Clinical Immunology and Allergy,
Department of Medicine, McMaster University, Hamilton, ON, Canada,
2
Division of Clinical Immunology & Allergy, McMaster University, Hamilton, ON, Canada.
RATIONALE: Eosinophils are multi-functional leukocytes involved in a
number of infectious and inflammatory processes, including allergic
diseases. Classical TH1 (IFNg) and TH2 (IL-4) cytokines negatively impact
eosinophil differentiation from cord blood (CB) in vitro. Since we have
recently shown that CB progenitor cells produce cytokines that auto-regulate their own differentiation, and we have recently uncovered a novel role
for TNFa positively impacting eosinophil-basophil (Eo/B) colony formation, we investigated the secretion of TNFa relative to these classical TH1
and TH2 cytokines during eosinophil differentiation from cord blood.
METHODS: Fresh CB progenitor cells were stimulated with GM-CSF,
IL-5 and IL-3 for 14 days in liquid media to induce eosinophil production.
Cells were isolated at days 3, 7 and 10 and cytokine levels of IL-4, TNFa,
IFNg, were evaluated using luminex technologies. Morphological analysis
was performed to determine the presence of eosinophils in culture after 14
days.
RESULTS: As compared the TH1 cytokine IFNg, TNFa was secreted at
significantly greater amounts at day 3 (p50.033), day 7 (p50.040), and
day 10 (p50.040) of culture. When compared to the TH2 cytokine IL-4,
TNFa was secreted at significantly greater amounts at day 3 (p50.014),
day 7 (p50.013), and day 10 (p50.019) of culture.
CONCLUSIONS: We show for the first time that CB Eo/B progenitors
preferentially secrete TNFa, compared to classic TH1 and TH2 cytokines,
during their own differentiation process. Our work provides novel insights
into cytokine signatures utilized by CB progenitors that may enhance
eosinophilic-basophilic inflammation in early life.

MONDAY

721