Binding Assay Of Human Papillomavirus-11 And Primary Human Respiratory Epithelial Cells

Objective. To establish an efficient method for direct isolation and culture of primary human respiratory epithelial cells from normal tissues, and investigate the attachment of human papillomavirus-11 (HPV-11) to respiratory epithelial cells, and discuss the effect of cellular heparan sulfate on virus-cell interaction.Methods.1. Human respiratory epithelial cells were isolated by 0.25%trypsin-EDTA, and cultured in a serum-free medium, and induced ciliated in an air-liquid interface system.2. The binding of HPV-11 virus-like particles (VLPs) to respiratory epithelial cells was detected by immunoblot, and quantified by the absorbance of bound particles measured by ELISA. The absorbance of squamous and ciliated cells was quantitively analysed using SPSS software.3. We further investigated the effect of heparin and heparitinase on the binding of HPV-11 virus-like particles (VLPs) to respiratory epithelial cells.Results.1. Primary laryngeal epithelial cells adhered to the cell flask after 6-7 days and formed a characteristic epithelioid shape showing a pavement-like arrangement. The epithelial phenotype of cells was also confirmed by expression of cytokeratins 5 and 8 on cell cytospins by immunofluorescence. Respiratory epithelial cells were observed with cilia appearance two weeks after air-liquid interface culture.2. HPV-11VLPs were capable of binding to surface of respiratory epithelial cells. There was no significant difference in the binding of HPV-11 to suamous and ciliated cells.3. The binding of HPV-11 and respiratory cells was reduced by heparin and heparitinase, and was in direct proportion with the concentration.Conclusions.1. The application of serum-free medium and air-liquid interface system in primary culture of human respiratory epithelial cells was effectively efficient.2. The disparity of HPV infection in upper airway was not caused by virus-cell attachment.3. The binding of HPV and respiratory epithelial cells required cellular heparan sulfate that possibly was the receptor of HPV.