In situ localization of infectious bursal disease virus-binding cells by a biotin-streptavidin system

Abstract

A biotin-streptavidin system was established to directly visualize infectious bursal disease virus (IBDV)-binding cells in cell culture or in fresh tissues. The cells or tissue sections were first incubated with a biotinylated, purified IBDV strain GZ911 and then with a streptavidin-β-galactosidase conjugate. In the presence of the enzyme substrate X-gal, IBDV-binding cells were labeled in blue color. By applying this method to frozen tissue sections, virus-binding sites were localized in situ in the bursa, spleen, and kidney tissue sections, whereas no positive cells were detected in the thymus tissue sections. Chicken embryo fibroblasts, Veto cells, MOP-8 cells, 293-EBNA cells, PANC-1 cells, and HuTu 80 cells were found to bind to the virus. However, the binding of the virus to MDA-MB-231 cells and SVG p12 cells was undetectable. This method can be employed for the expressional cloning of IBDV receptor and can be applied to studies on other avian viruses.