Phenotyping and Cytokine Regulation of the BEAS-2B Human Bronchial Epithelial Cell: Demonstration of Inducible Expression of the Adhesion Molecules VCAM-1 and ICAM-1

Abstract

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.

Transcriptional Mechanisms Regulating Alveolar Epithelial Cell-specific CCL5 Secretion in Pulmonary Tuberculosis

“…Epithelial cells, covering the entire alveolar surface area of the lung of ϳ70 m 2 (12), may contribute to host defense by chemokine production (13), by adhesion molecule expression (14), and possibly by antigen presentation via HLA-DR expression (15). CCL5 may be secreted by pulmonary epithelial cells in response to proinflammatory cytokines (5,13,16,17), hyperosmolarity (18), or pathogens such as RSV (19,20).…”

TNF-α Potentiates C5a-Stimulated Eosinophil Adhesion to Human Bronchial Epithelial Cells: A Role for α5β1 Integrin

“…Despite a number of studies, including our own, that have addressed the subject of eosinophil adhesion to epithelial cells (11)(12)(13)(14)(15), the adhesion pathways governing these interactions remain to be fully elucidated. It is known that eosinophils bind ICAM-1 expressed on HBEC predominantly via the ␤ 2 integrin, ␣ M ␤ 2 (CD11b/CD18) (14,15).…”

Expression and function of beta1 integrins on human eosinophils

“…Until recently, only ICAM-1, but not E-selectin or VCAM-1, had been identified in the respiratory epithelium in vitro and in in vivo biopsies from patients with asthma (Bloemen et al 1993, Fukuda et al 1996, Stark et al 1996. However, in the BEAS-2B bronchial epithelial cell line, culture with TNF or IL-1 was found to induce VCAM-1 mRNA and cell surface expression (as well as ICAM-1 expression), while culture with IL-4 induced VCAM-1 but not ICAM-1 expression (Atsuta et al 1997). Maximal VCAM-1 expression resulted from the combination of TNF and IL-4.…”

“…Unlike neutrophils, eosinophils express the α4 integrins VLA-4 (α4β1) and α4β7 which mediate binding to VCAM-1, an immunoglobulin superfamily member induced by cytokines on endothelium and epithelial cell lines (Atsuta et al 1997, and to an alternatively spliced domain in fibronectin, CS-1 (Anwar et al 1994. The α4β7 integrin binds to the mucosal addressin cell adhesion molecule-1 (MAdCAM-1) that has structural homology to ICAM-1 and VCAM-1 (Walsh et al 1996, Briskin 1997.…”