Terri H. Beaty and Shau-Ku HuangSchleimer, Luis Caraballo, Raana P. Naidu, Paul N. Levett, Freidhoff, Claudia Sengler, James R. Plitt, Robert P.Beyer, Kathleen C. Barnes, Beverly S. Plunkett, Linda R. Renate G. Nickel, Vincenzo Casolaro, Ulrich Wahn, Kirstenhttp://www.jimmunol.org/content/164/3/1612J Immunol€2000; 164:1612-1616; ;Referenceshttp://www.jimmunol.org/content/164/3/1612.full#ref-list-1This article cites 29 articles, 14 of which you can access for free at: Subscriptionshttp://jimmunol.org/subscriptionsInformation about subscribing to The Journal of Immunology is online at: Permissionshttp://www.aai.org/ji/copyright.htmlSubmit copyright permission requests at: Email Alertshttp://jimmunol.org/cgi/alerts/etocReceive free email-alerts when new articles cite this article. Sign up at:

Rationale Since respiratory RNA viruses are an important cause of exacerbations of sinusitis and asthma, we tested the ability of human rhinovirus-16 (HRV-16) and the toll-like receptor type 3 (TLR3) agonist, dsRNA, to alter expression of B7 homolog costimulatory molecules in human airway epithelial cells. Methods BEAS2B human airway epithelial cells were cultured to confluence, exposed to dsRNA (25 μg/ml), and then expression of mRNA and cell-surface protein for B7 homologs was assessed by real-time PCR and flow cytometry, respectively. Additionally, human subjects were infected with HRV-16 in vivo, and mRNA for B7-homologs was assessed by real-time PCR in fresh nasal epithelial cell scrapings obtained before and daily up to 4 days. Results Exposure of BEAS2B cells to dsRNA for 24 hours resulted in an increase in cell-surface and mRNA expression of B7-H1 (approx. 2 and 3 fold, respectively) and B7-DC (approx. 2.5 and 3.5 fold), but not B7-H2 or B7-H3 (p Conclusion Increased expression of the B7 homologs B7-H1 and B7-DC by exposure of epithelial cells to the TLR3 agonist dsRNA or HRV-16 may influence the development of adaptive immune responses in the airways.

Chemokine-induced eosinophil chemotaxis is mediated primarily through the C-C chemokine receptor, CCR3. We have now detected CCR3 immunoreactivity on epithelial cells in biopsies of patients with asthma and other respiratory diseases. CCR3 mRNA was detected by Northern blot analysis after TNF-α stimulation of the human primary bronchial epithelial cells as well as the epithelial cell line, BEAS-2B; IFN-γ potentiated the TNF-α-induced expression. Western blots and flow cytometry confirmed the expression of CCR3 protein. This receptor is functional based on studies demonstrating eotaxin-induced intracellular Ca2+ flux and tyrosine phosphorylation of cellular proteins. The specificity of this functional response was confirmed by blocking these signaling events with anti-CCR3 mAb (7B11) or pertussis toxin. Furthermore, 125I-eotaxin binding assay confirmed that CCR3 expressed on epithelial cells have the expected ligand specificity. These studies indicate that airway epithelial cells express CCR3 and suggest that CCR3 ligands may influence epithelial cell functions.

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Airway epithelium may actively participate in inflammatory responses, such as occur in asthma. The presence and regulation of surface molecules on the airway epithelium, however, is incompletely understood. We have determined the phenotype of the human bronchial epithelial cell line BEAS-2B by flow cytometry. We confirmed previous observations that human bronchial epithelial cells constitutively express CD29, CD44, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD51, CD54 (ICAM-1), CD61, and HLA class 1. BEAS-2B cells were also found to constitutively express CD9, CD13, CD15, CD15s, CD23, CD33, CD36, CD40, CD41b, CD42b, CD48, CD50, CD71, and CD102 (ICAM-2). Culture of BEAS-2B cells with tumor necrosis factor (TNF)-alpha or interleukin (IL)-1beta (1 ng/ml) was found to enhance intercellular adhesion molecule-1 (ICAM-1) expression (several fold) and induce de novo CD106 [vascular cell adhesion molecule-1 (VCAM-1)] expression. TNF-alpha or IL-1beta did not change the expression of CD9, CD13, CD16, CD23, CD29, CD31, CD32, CD35, CD45, CD61, or CD64 in BEAS-2B cells. IL-4 (1 ng/ml) also induced expression of VCAM-1 (1.5-fold) but not ICAM- expression while interferon-gamma (1 ng/ml) enhanced only ICAM-1 expression (2-fold). Maximal VCAM-1 expression was obtained with the combination of TNF-alpha and IL-4 (8-fold). Using Northern blot hybridization analysis, ICAM-1 and VCAM-1 mRNA was detected in BEAS-2B cells stimulated with cytokines. VCAM-1 on stimulated BEAS-2B was functionally active as determined by adhesion of purified eosinophils and blockade with specific antibodies. Primary isolates of bronchial epithelial cells produced detectable levels of VCAM-1 protein and mRNA as detected by enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction, respectively. These results suggest that cytokine activation induces expression of ICAM-1 and VCAM-1 on airway epithelium, an event which may influence leukocyte infiltration and activation.