486 Rhinovirus (RV) 16 stimulation of interferon (IFN)-α and -γ by human peripheral blood mononuclear cells (PBMC)

Abstracts

J ALLERGY CLIN IMMUNOL VOLUME 105, NUMBER 1, PART 2

an element mediating antigen-induced IL-4 expression in Th2 cells via binding of the Ca*+- and calcineurin-dependent factor(s) NFAT. We therefore studied the effects of salicylates on activation of the

487

NFAT family member NFAT-I in PBT clones generated by priming with 5 pglml PHA and maintained in media supplemented with 5 U/ml IL-2. Cells were treated with ASA (IO-3 M) or DMSO vector

pahtogenesis of asthma. In the present study, we analyzed the functional role of the c-kit ligand. stem cell factor (SCF), an important activating and chemotactic factor for mast cells and eosinophils, in the pathogenesis of asthma. We could suppress SCF expression in

induced the rapid (within 30 min) and transient translocation of NFAT-1 into the nuclei of most cells. Interestingly, treatment with ASA caused more sustained nuclear staining for NFAT- I in PBT. due

NIH 3T3 and SPI epithelial cells in vitro and murine dermis in vivo by a specific antisense phosphorothioate oligonucleotide overlapping the translation start site of SCF, whereas control oligonucleotides

to delayed cytosolic re-export of the factor. Increased nuclear expression of NFAT-I in ASA-treated PBT was apparent at 60 and 90 min post-stimulation by immunofluorescence, as well as by Western blots

were virtually inactive. We then focused on the role of SCF in a murine model of asthma associated with late phase allergic pulmonary inflammation in ovalbumin (OVA)-sensitized mice: Local intranasal administration of FITC-labelled SCF antisense oligonu-

and DNA-binding assays using nuclear extracts from parallel samples. We conclude that the inhibitory effects ofASA and related compounds on IL-4 expression in human T cells cannot be accounted for by reduced NFAT-I activation. Although sustained NFAT-I nuclear

cleotides led to strong DNA uptake in interstitial lung cells associated with a striking reduction of intacellular SCF expression. Such intrapulmonary blockade of SCF expression caused an abrogation of sign

translocation can be invoked to explain the enhancing effects of salicylates on a minimal (9%bp) IL4 promoter construct, the molecular basis for their overall inhibitory effects on IL-4 expression remains to

of lung inflammation including mast cell activation, TH2 cytokine production, infiltration of eosinophils and bronchoconstriction. SCF antisense treatment in OVA-sensitized mice was at least as effective

be determined.

Rhinovirus (RV) 16 Stimulation of Interferon (IFN) -a and -y by Human Peripheral Blood Mononuclear Cells (PBMC) J bring, R Vrris,

C Swenson.

J Gem.

W Busse

University

respiratory viruses is important and generating proinflammatory In previous studies (J Immunol

as treatment with corticosteroids. These data indicate a critical role for SCF in a murine asthma model and suggest that anitsense oligonucleotides to SCF may be a novel selective approach for the treatment

of Wisconsin,

Madison, WI Viral respiratory infections, particularly RV. are important causes of asthma exacerbations. It is proposed that the immune response to in eliminating the infectious pathogen cytokines to enhance airway injury. 1996; 157: 1605). we used an in vitro

Local Administration of Antisense Phasphorothioate Oligonucleotides to the C-Kit L&and, Stem Cell Factor, Abrogates Airway Inflammation and Suppressed TH2 Cytokine Production in a Murine Model of Asthma S Finorto, M Buerke, K Lindau, R Atreya, S Wirtz. E Schmitt, PR Galle, MF Neurath Laboratory of Immunology, I Medical Clinic, University of Mainz, Germany Mast cells and eosinophils appear to play a fundamental role in the

prior to stimulation with 0.25 l,tg/ml A23187. and NFAT-I activation was analyzed by immunofluorescent staining using specific antibodies. In agreement with previous reports, Ca*+-mediated stimulation

486

S159

of inflammatory

488

disorders

such as asthma

in humans.

Lung Delivery of an Interleukin-9 Antibody Tkatment Inhibits Airway Hyperresponsiveness (AHR), BAL Eosinophilia, Mu& Production, and Serum IgE Elevation to Natural Antigens in a Murine Model of Asthma MP M&me*. J Tepperf, C Weiss*, Y

model system to define the immune response to RV and found RV16 activated lymphocytes (increased CD69 expression) and stimulated IFN-1 production, which was a monocyte-dependent response. To

Tamer*, laides*, Meeting,

determine whether RV 16 activation of monocytes generated IFNa( to cause lymphocytes to produce IFN-y production by RV16, PBMC were obtained from subjects who had had a previous infection with

We have shown previously that interleukin-9 (IL9) is an important regulator of bronchial responsiveness and allergic inflammatory responses in the lung, and has been genetically linked to asthma and

RV 16 and then incubated with infectious RV 16 for 48 hours. Following this incubation, the cells were pelleted and supemates used to measure IFN-a and IFN-yproduction, two cytokines involved in the

allergy in humans. In order to study the effects of specific blockade of IL9 on the lung inflammatory response to natural antigens, high affinity >I nmolar) monoclonal anti-IL9 antibody was administered via

immune response to respiratory viruses (Table; n = 7; mean ( SE). These observations indicate: (1) RV16 stimulates production of IFN-a and IFN-y. (2) the IFN response to RV 16 is ICAM- 1 receptor

the tracheal instillation during the course of antigen exposure. Cohorts included mice treated with isotype matched control antibody, and untreated animals. Intranasal antigen was given over 22-days

dependent; and promotes IFN-y not a significant response to RV

(3x/wk). Approximately 12 hrs after the last antigen challenge, mice were anesthetized, airway responsiveness was measured, and BAL and blood were obtained. Lungs were harvested, fixed in 10% NBF, processed and graded for histopathology. Antigen exposure signifi-

(3) neutralization of IFN-a significantly (p ~0.05) production. These findings suggest IFN-a alone is inducer of IFN-a. It may, however, be that the IFN-a I6 is an important determinant of IFN-1 production.

Although the mechanisms of this interaction mined, it is possible that an absence of IFN-a longed RV16 survival and hence compensatory

have not been deteris associated with proactivation of IFN-p

Moreover, abnormalities in IFN-a production may, therefore, affect IFN-yproduction, its influence on virus-host defense, and the development of asthma. [Supported by an NIH grant, A1348911 IFN-a

(&ml) RV16+

RV16+

RV16+

anti-

anti-

anti-

ICAM-I

IFN-a

Control

RV16

ICAM-I

Control

RV16

Not

11.600

Not

7.5

76.6

96

280

Detected

2961

Detected

k2.4

+27.9

24.0

*94

RE Taylorf;, D Tumasf, Y Zhou*, A Ha&u*, RC Lmirt* *Magainin Pharmaceuticals, Inc. PA tGenentech Inc. South San Francisco, CA

NC NicoPlymouth

cantly increased AHR (I.7-fold), BAL eosinophils (25,000 cells/lung), serum total IgE (2.4-fold), and the overall histopathology grade compared to control mice. Anti-IL9 treatment significantly inhibited antigen-induced AHR (P ~0.05). BAL eosinophilia, serum IgE (P ~0.05). and histopathology grade (P 4.05) compared to sensitized untreated or control antibody treatment. Qualitative analyses of mucin production also demonstrated normalization with anti-IL9 antibody treatment. Significant improvement in all measures of the lung allergic inflammatory response by the specific blockade of IL9 demonstrate a broad functional role for IL9 in their pathogenesis. Moreover, our results suggest that the direct administration lung of IL9 blocking antibody may provide clinically significant efit in allergic inflammatory diseases, such as asthma.

to the ben-