HSV INFECTION OF POLARIZED EPITHELIAL-CELLS ON FILTER SUPPORTS - IMPLICATIONS FOR TRANSPORT ASSAYS AND PROTEIN LOCALIZATION

Epithelial cell lines can be grown on filter supports and form polariz ed monolayers with distinct basolateral and apical plasma membrane dom ains. This property has been extensively used in cell biology to inves tigate epithelial cell function. To date, a major limitation of this a pproach was been the difficulty of obtaining transient gene expression in polarized epithelia. Here we present an approach to overcome this problem using gene transfer into polarized epithelial cells grown on f ilters using a herpes virus-based vector. Recombinant genes are insert ed into a defective HSV-1 plasmid and packaged with a replication-inco mpetent HSV-1 helper virus into virus particles which are used to infe ct the polarized epithelial cells grown on filters. The transepithelia l resistance of the cells is not affected by the addition of virus, an d there are no detectable cytopathic effects.