Recovery of an adenovirus from a feral rodent Peromyscus maniculatus.
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Biomedical subjects
Publications and source records attributed to W P Rowe.
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The titers of adenovirus and SV40 genetic carriers in hybrid preparations (E46(+)) can be quantitated by determining the percentage of cells showing neoantigens stainable with fluorescent antibody (FA) at 21 to 24 hours; both titers can be obtained with a single coverslip. The adenovirus and SV40 antigen-inducing titers so obtained are of the same order of magnitude in stock preparations of E46(+) grown in either African green monkey kidney (AGMK) or human embryonic kidney tissue culture (HEK), and are generally within one log of the infectivity titer. Quantitative studies of 50 degrees C heat inactivation, ultracentrifugation, and equilibrium density gradient centrifugation of E46(+) gave no indication that SV40 neoantigen induction could be dissociated from adenovirus, whereas adenovirus and SV40 virus grown as a mixed infection were readily dissociated by these procedures. Pretreatment of HEK cells with a medium containing 5-fluorouracil desoxyriboside (FUDR) did not affect induction of either adenovirus or SV40 neoantigen or development of cytopathic effects after infection with E46(+), but did prevent formation of Ad. 7 viral antigen.
E46(+) virus purified by limiting dilution and plaque isolation in AGMK cells consists of a mixture of hybrid and non-hybrid virus particles. Of 102 plaque isolates obtained in HEK cells, 98 contained no detectable hybrid virus, while 4 contained a small proportion of hybrid virus. Plaque isolates from AGMK cells uniformly induced SV40 neoantigen, but when these plaque isolates were plaqued in HEK cells the progeny were again non-hybrid adenovirus. It thus appears impossible to obtain a pure line of hybrid virus. The proportion of hybrid virus produced in HEK cells fell progressively with decreasing multiplicity of infection, and plaque induction by E46(+) virus in AGMK followed 2-hit kinetics, indicating that the hybrid virus particles are defective and require dual infection with non-hybrid virus for their propagation.
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A passage line of a spontaneous hamster fibrosarcoma is contaminated by the virus. of lymphocytic choriomeningitis. Tumors from animals receiving implants when newborn contain high titers of infectious lymphocytic-choriomeningitis virus and complement-fixing antigen, and hamsters receiving implants when weanlings develop high titers of complement-fixing antibody against lymphocytic-choriomeningitis virus. In contrast with the specific reactions of tumorous hamsters to the initiating virus in virus-induced tumors, the development of complement-fixing antibody to lymphocytic-choriomeningitis virus does not depend on the development of tumors. Infant hamsters bearing the tumor have a generalized subclinical infection and seem able to spread virus to other hamsters and to humans.
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