Abstract
Purified type 5 adenovirus was disrupted at pH 10.5 and the capsid shown to be comprised of two characteristic morphological subunits: (a) Hollow, polygonal structures corresponding to the virus capsomeres seen in preparation of purified virus and (b) thread-like strands also identifiable in preparations of purified virus. These structures were compared morphologically with purified preparations of the group- and type-specific soluble antigens characteristically produced in mammalian cells infected with adenoviruses. The group-specific soluble antigen was a homogeneous preparation of hollow, polygonal rods identical with the virus capsomeres. The type-specific soluble antigen corresponded to the thread- or fiber-like components of the purified virus particle. Inspection of disrupted virus preparations confirmed earlier immunological data which indicated that the major virus antigen was the group-specific soluble antigen. These data provide convincing evidence for the hypothesis that the adenovirus-induced soluble antigens represent virus subunits produced in excess.
Highlights
Materials and MethodsPreparation of Purified and DegradedVir~.--Highly purified (3 times CsCl-banded) type 5 adenovirus was prepared as described in the preceding paper [2]
Purified type 5 adenovirus was disrupted at p H 10.5 and the capsid shown to be comprised of two characteristic morphological subunits: (a) Hollow, polygonal structures corresponding to the virus capsomeres seen in preparation of purified virus and (b) thread-like strands identifiable in preparations of purified virus
These structures werc compared morphologically with purified preparations of the group- and type-specific soluble antigens characteristically produced in mammalian cells infected with adenoviruscs
Summary
Preparation of Purified and DegradedVir~.--Highly purified (3 times CsCl-banded) type 5 adenovirus was prepared as described in the preceding paper [2]. Prior to electron microscopy these preparations were dialyzed against 0.01 ~ phosphate buffer, pH 7.5, and assayed for infectivity and complement-fixing activity as described previously (s). A mixture of type-specific (E) antigen and toxin (E-T) was obtained from DEAE columns. This latter material was not contaminated by L antigen or normal host cell materials. The final material obtained consisted of a mixture of E and L antigens and toxin. Prior to use, these preparations were dialyzed against 0.01 ~ phosphate buffer at pH 7.5 and the quantity of infectious virus, complementfixing antigens, and toxin determined by appropriate titration procedures [3]. Assays.--Infectivity, toxin, and complement-fixation titrations were done as described previously [2]
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