Altered trafficking of cells during acute pulmonary allergic and viral inflammatory stimuli in rats...

187 Prostaglandin D2-Induced Eosinophilic Airway InflammationIs Mediated by CRTH2 Receptor

Y. Shiraishi1,2, K. Asano1,2, T. Nakajima1,2, K. Niimi1,2, Y. Suzuki1, T.Shiomi1, K. Sayama1, T. Oguma1, E. Ikeda3, H. Hirai4, K. Yamaguchi1, A.Ishizaka1; 1Division of Pulmonary Medicine, Department of Medicine,Keio University, Tokyo, JAPAN, 2Pfizer-Keio Research Laboratories,Tokyo, JAPAN, 3Department of Pathology, Keio University, Tokyo,JAPAN, 4R&D Center, BioMedical Laboratories, Saitama, JAPAN.RATIONALE: Prostaglandin D2 (PGD2) is one of the key modulators ofallergic airway inflammation. PGD2 exhibits direct chemotactic activityfor eosinophils via CRTH2 (DP2) receptor in vitro, however, the role ofCRTH2 for the in vivo eosinophil trafficking into the airway has not beendetermined yet. We thus examined the role of CRTH2 on PGD2-inducedairway eosinophilia in vivo.METHODS: PGD2 or CRTH2/DP/TP receptor-specific agonists wereadministered intratracheally in BN rats pretreated with systemic inter-leukin 5 (IL-5). The degree of airway eosinophilia was evaluated by dif-ferential cell count of bronchoalveolar lavage (BAL) fluid and lung histol-ogy.RESULTS: Peripheral blood eosinophilia was induced by IL-5 pretreat-ment alone, and administration of PGD2 exhibits no additional effects. Incontrast, airway eosinophilia was not induced by IL-5 pretreatment alone,while intratracheal PGD2 administration in IL-5 pretreated animalsinduced a robust BAL eosinophilia accompanied by perivascular and peri-bronchial eosinophil infiltration. CRTH2 receptor-specific agonists such asDK-PGD2 and indomethacin exhibited an equivalent effect on airwayeosinophilia as PGD2, but a DP receptor agonist, BW 245C, or a TP recep-tor agonist, I-BOP, did not. The PGD2- or CRTH2 receptor agonist-inducedairway eosinophilia was significantly inhibited by the pretreatment with aCRTH2/TP dual receptor antagonist, ramatroban, but not by a TP receptorantagonist, SQ29,548, or a DP receptor antagonist, BW A868C.CONCLUSIONS: CRTH2 receptor mediates in vivo chemotactic activi-ty of PGD2 for eosinophils.

188 Evolution of Aspirin Hypersensitivity in Allergic and Non-Allergic Types of Rhinitis

E. Nizankowska, M. Swierczynska, A. Szczeklik; Med. Dept. of Jagiel-lonian University School of Medicine, Cracow, POLAND.RATIONALE: Although both non-allergic rhinitis with eosinophilia syn-drome (NARES) and allergic rhinitis (AR) were suggested to precedeaspirin hypersensitivity, no prospective study aiming to detect abnormalresponses to aspirin in NARES or AR was yet conducted.METHODS: Thirty subjects with NARES and thirty with AR, without his-tory of aspirin hypersensitivity, underwent single-blind, placebo-controlledoral challenge with aspirin, assessed by clinical symptoms, spirometry,acoustic rhinometry and rhinomanometry. Urinary leukotriene E4 (u-LTE4), plasma (p-) and urinary (u-) 9alpha,11beta-prostaglandin F2(9alpha,11beta-PGF2) were analyzed by ELISA and gas chromatogra-phy/mass spectophotometry, respectively, at baseline and following aspirinchallenge.RESULTS: In NARES all aspirin challenges were negative. In 5 of ARsubjects aspirin induced a spectrum of hypersensitive responses varyingfrom bronchial and/or nasal ones (n=2) to asymptomatic biochemicalalterations only (n=3). They had higher mean baseline levels of u-LTE4than other subjects with AR or NARES (967.4±753 vs. 524.5±789.5 and520±461.8 pg/ml creat.), whereas basal p-9alpha,11beta-PGF2 levelswere similar. An increase in u-LTE4 (p<0.05) and p-9alpha,11beta-PGF2(p=0.07) following aspirin was observed only in subjects with AR andabnormal reaction to aspirin. They were characterized by a more severerhinosinusitis, and four of them had concomitant bronchial asthma. Theaspirin challenges repeated two years later revealed nasal responses in 2out of 3 subjects with AR who previously had responded only withasymptomatic biochemical alterations.CONCLUSIONS: The individual cases of persistent and severe allergicrhinitis may be associated with early phase of aspirin hypersensitivity,characterized by eicosanoid metabolism alterations.

J ALLERGY CLIN IMMUNOL Abstracts S47VOLUME 115, NUMBER 2

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and Viral Inflammatory Stimuli in Rats With the Chronic Post-bronchiolitis Asthma-Like Phenotype

R. L. SORKNESS1, L. A. ROSENTHAL2, T. PIER2, S. P. AMINEVA2,R. J. SZAKALY2, R. F. LEMANSKE, Jr.3; 1Sch of pharmacy, UNIVER-SITY OF WISCONSIN, Madison, WI, 2Medicine, UNIVERSITY OFWISCONSIN, Madison, WI, 3Pediatrics, UNIVERSITY OF WISCON-SIN, Madison, WI.RATIONALE: Airway inflammation in asthma may be due to enhancedvulnerability of airways to common inflammatory stimuli. We hypothe-sized that rats with chronic airway dysfunction would differ from normalsin their acute pulmonary inflammatory responses. METHODS: Normal BN rats were compared with asthma-like BN ratsthat were >10wks post early age bronchiolitis. Acute allergic inflamma-tion was induced by a single exposure to aerosolized ragweed pollenextract 2wks after sensitization, and evaluated at 1 or 7 days; viral inflam-mation was induced with a tracheal insufflation of attenuated mengovirus,and evaluated in 3 days. Airspace infiltrates were quantified with bron-choalveolar lavage (BAL), and airway wall eosinophils (eos) with stain-ing of major basic protein (MBP) in lung sections. RESULTS: Unchallenged normal and asthma-like rats did not differ sig-nificantly with regard to BAL eos or airway MBP. However, the asthma-like rats had significantly greater airway wall MBP (p=0.001) and fewerBAL eos (p=0.02) 1 day after allergen challenge, and fewer BAL eos(p=0.04) with similar airway MBP 7 days after allergen. During viralinfection a similar pattern occurred, the asthma-like rats having few eosand neutrophils in BAL (p<0.04), but present in lung sections. BAL viraltiters were similar in the 2 groups. CONCLUSION: During acute inflammation the asthma-like rats have fewereos present in the airspace, but equal or greater numbers in airway walls, com-pared with normals. These data are consistent with the hypothesis that asth-matic airways exhibit different patterns of inflammatory cell trafficking thatresult in greater vulnerability of the airways to allergic or viral insults.Funding: NIH-NHLBI

190 Specific Induction of RELM-� in a Murine Asthma Model

S. Kierstein, A. Haczku, S. A. Keilbaugh, G. Vass, X. Yang, I. G. Redai,G. D. Wu; University of Pennsylvania, Philadelphia, PA.RATIONALE: Resistin-like molecule (RELM)-� a novel cysteine-richsecreted protein has recently been shown to be activated by Th2 mediatedimmune responses in the gut. The aim of this study was to determine theinvolvement of this molecule in the pathogenesis of the allergic airwayresponse and to compare its expression pattern with the pulmonary surfac-tant protein (SP)-D, known to be secreted by epithelial cells of the airways.METHODS: Balb/c mice were sensitized and challenged withAspergillus fumigatus (Af). Sequential studies of lung function, inflam-matory cell influx and cytokine levels were performed 12, 24 and 48hafter intranasal provocation. Expression of the pulmonary surfactant pro-tein (SP)-D and RELM-� was investigated by Western blot. mRNAexpression changes in the lung were assessed using pathway-specificmicroarrays and Northern blot analysis.RESULTS: Increase in airway obstruction in sensitized and challengedanimals was paralleled by a massive influx of neutrophils at 12h andeosinophils at 24-48h after challenge. mRNA activation and release ofproinflammatory cytokines IL-4 (532.5 pg/ml), IL-6 (315.5 pg/ml) and IL-5 (186.2 pg/ml) also occurred within 24h after allergen provocation. Theheight of pro-inflammatory changes was followed by a resolution 48 hoursafter Af challenge of sensitized mice. Interestingly, levels of the potentimmunomodulatory SP-D reached their peak also at the 48h time point par-alleled by the peak of RELM-� protein expression in the BAL fluid.CONCLUSIONS: The structural similarities and similar expression pat-terns of SP-D and RELM-� may indicate coordinated regulation and/orfunction for these novel proteins during the resolution of allergen inducedinflammatory changes.Funding: R01 AI055593-01, ALA RG144N