Herpes Simplex Virus Products in Productive and Abortive Infection I. Stabilization with Formaldehyde and Preliminary Analyses by Isopycnic Centrifugation in CsCl

Abstract

Lysates of HEp-2 cells productively infected with herpes simplex virus yielded two bands on isopycnic centrifugation in CsCl gradients, ranging from 1.2 to 1.6 g/cm 3 . One band, designated α, had a mean buoyant density of 1.27 g/cm 3 and contained herpes virions. Band β had a mean density of 1.305 g/cm 3 and contained primarily complement-fixing viral antigens and little or no viral deoxyribonucleic acid (DNA). The products banding in the α and β bands were unstable; fivefold or higher amounts were recovered by treating the cell extract with formaldehyde prior to centrifugation. Formaldehyde treatment increased the buoyant density of viral products in both the α and β bands by about 0.015 g/cm 3 . In addition, it stabilized hitherto inapparent products, forming a broad band γ with a density range of 1.37 to 1.45 g/cm 3 . The material in the γ band was heterogeneous; it contained viral DNA, cellular DNA, and viral antigen. Formalinized lysates of DK cells abortively infected with herpes simplex virus yielded a β band undifferentiated from that formed by extracts of productively infected cells. The γ band was less dense and narrower. The α band was entirely missing.