Analysis of VSV mutant with attenuated cytopathogenicity: Mutation in viral function, P, for inhibition of protein synthesis

SUMMARYWild-type (wt) strains of vesicular stomatitis virus (VSV) strain Indiana are poor to non-inducers of interferon (IFN) which express IFN induction-suppressing activity. At non-permissive temperatures, temperature-sensitive (ts) mutants of this virus are either like their wt parents, or they are good to excellent inducers of IFN. IFN inducibility and IFN induction-suppressing activity are mutually exclusive phenotypes in VSV-Indiana. With one exception, all Orsay ts mutants derived by A. Flamand (CNRS, Gif-sur-Yvette, France), representing the five complementation groups, were poor to non-inducers of IFN and were also capable of suppressing IFN induction by other viruses. In contrast, all Glasgow ts mutants derived by C. R. Pringle (University of Warwick, Coventry, U.K.) were excellent inducers of IFN. We demonstrate that this difference in acquisition of IFN inducibility relates primarily to the origin of the mutations; spontaneous for Orsay, and mutagen-derived for Glasgow. Tests with newly generated spontaneous and mutagen-derived mutants, and temperature-stable revertants of IFN-inducing ts mutants indicate that IFN inducibility results from non-ts, multiple mutations rarely acquired spontaneously, but generated frequently upon mutagenesis with 5-fluorouracil. The capacity of VSV-Indiana to induce IFN is considered intrinsic to the virus, but is only manifested when the dominant IFN induction-suppressing phenotype is lost through mutagenesis. Thus, non-ts mutations appear to regulate the expression of the IFN induction-suppressing phenotype, and hence the IFN inducibility of VSV-Indiana.

Section: Discussionmentioning

confidence: 99%

Interferon Induction by Viruses. XVII. Non-temperature-sensitive Mutations Regulate Interferon Induction by Vesicular Stomatitis Virus

Sekellick

Marcus²

1989

“…Vesicular stomatitis virus (VSV) T 1026R1 is a non-ts revertant of a mutant originated by C. P. Stanners (Stanners et at., 1977). Stock VSV T1026R1 was prepared from plaque-purified virus in Vero cells.…”

Section: Discussionmentioning

confidence: 99%

“…Mouse L-929 fibroblasts and Vero cells were originally obtained from Flow Laboratories. They were grown in Eagle's minimum essential medium (MEM) containing 10~ newborn bovine serum (NBS) and maintained in MEM plus NBS, 2~ for L-929 and 5~ for Vero cells.Vesicular stomatitis virus (VSV) T 1026R1 is a non-ts revertant of a mutant originated by C. P. Stanners (Stanners et at., 1977). Stock VSV T1026R1 was prepared from plaque-purified virus in Vero cells.…”

mentioning

confidence: 99%

Distinctive Characteristics of Crude Interferon from Virus-infected Guinea-pig Embryo Fibroblasts

Winship

Fong

Hsiung

1984

SUMMARYCrude interferon preparations from primary guinea-pig embryo cells infected with vesicular stomatitis virus strain T1026R1 were shown to be more sensitive to heat (37 °C), pH 2.0, and SDS than crude mouse interferon. Since the proportion of antiviral activity lost after each treatment was nearly the same, the existence of a single fraction of antiviral activity sensitive to all three treatments was suggested. Support for this possibility was given by the finding that subjecting this guinea-pig interferon to any one of the treatments rendered it insensitive to the effects of the other two.We have previously shown that guinea-pig embryo fibroblasts (GPE cells) infected with several different viruses were capable of producing more antiviral activity (interferon) than had been previously reported by other laboratories (Kaplan et al., 1962;Friedman et al., 1962;Lackovic et al., 1979;Warfel & Stewart, 1980;Sonnenfeld, 1981 ;Christofinis, 1980), but that much of this activity was too labile to be detected by conventional means (Winship et al., 1983). It was found that virus type, multiplicity of infection, and cell age are important factors in determining the total yield of these substances (Winship et al., 1983).In this communication, we describe several basic biological and physical characteristics of crude guinea-pig interferons, These were compared with crude, virus-induced mouse interferons with 'classical' characteristics. It was found that crude guinea-pig interferons were a mixture of conventional interferon and a heat-, pH 2-, and SDS-labile fraction constituting approximately 60~ of the antiviral activity.GPE fibroblasts were prepared from whole guinea-pig foetuses (30 days gestation) as described previously (Hsiung et al., 1976). Primary and passaged GPE cell cultures were grown in culture flasks or roller bottles for interferon induction and for assays as described by Winship et al. (1983). Mouse L-929 fibroblasts and Vero cells were originally obtained from Flow Laboratories. They were grown in Eagle's minimum essential medium (MEM) containing 10~ newborn bovine serum (NBS) and maintained in MEM plus NBS, 2~ for L-929 and 5~ for Vero cells.Vesicular stomatitis virus (VSV) T 1026R1 is a non-ts revertant of a mutant originated by C. P. Stanners (Stanners et at., 1977). Stock VSV T1026R1 was prepared from plaque-purified virus in Vero cells. Wild-type VSV, Indiana strain, was originally obtained from the American Type Culture Collection (no. VR-158). Assays of wild-type VSV could be conducted in Vero, GPE and L-929 cells with nearly equal efficiency.Guinea-pig interferons were produced in 32 oz culture flasks or roller bottles of confluent GPE cells. For interferon induction, confluent GPE cells were aged for 7 days in growth medium which was maintained at neutral pH by adding a small amount of sodium bicarbonate, without medium change. The aged cells were infected with VSV T1026R1 under conditions previously described (Winship et al., 1983) and fed with MEM containing 0.5 ~ NBS. The infected cultures ...

“…The titre of the revertant was 10 s p.f.u./ml. At low virus dilutions, no plaques could be seen, and this was shown to be due to the presence of defective interfering particles, whose presence was demonstrated after centrifugation on a velocity gradient (Stanners et al, 1977) (not shown).…”

Section: Effect Of Preinfection With Vsv (Ts Mutant 1026) On Rsv Exprmentioning

confidence: 99%

“…So as to be able to assess RSV expression, we chose systems which are non-permissive for the rhabdoviruses used: RV is not pathogenic for chicks and VSV ts mutant 1026 is a double mutant, non-cytopathogenic at the nonpermissive temperature (Stanners et al, 1977).…”

Section: Sarcoma Virus Does (Mouttet Et Al 1980) Both Influenza VImentioning

confidence: 99%

Inhibition of Rous Sarcoma Virus-induced Transformation by Preinfection with Rhabdoviruses

Semmel¹,

Sathasivam

1983

SUMMARYIn vivo preinfection of chicks with rabies virus (RV) or vesicular stomatitis virus (VSV) ts 1026 inhibits tumour formation after superinfection with Rous sarcoma virus (RSV). The degree of inhibition depends on the titre of the infecting viruses and the interval between rhabdovirus and RSV infection. In vitro, cells preinfected with VSV ts 1026 under non-permissive conditions and superinfected with RSV, are not transformed as judged by cell morphology, serum requirement for growth or the capacity to form colonies in soft agar, all these being the same as in uninfected cells. Doubly infected cells take up less deoxyglucose than cells infected with RSV only and more than cells infected with VSV only. RSV multiplication is inhibited in doubly infected cells: the supernatant fluid of these cells contains fewer focus-forming units and less reverse transcriptase activity than that of cells infected with RSV only. Doubly infected cells contain both VSV and RSV internal antigens 15 days after infection. The supernatant fluid of cells infected with VSV and maintained under non-permissive conditions inhibits transformation by RSV and multiplication of RSV, but not of VSV. Under non-permissive conditions, the rhabdoviruses undergo at least part of the infectious cycle, but no infectious virus is produced. RV antigen can be detected in the brain of parenterally infected chicks and VSV antigen in cells infected 15 days previously. We conclude that the inhibition of RSV multiplication and expression is probably due to one or more processes linked to the persistence of rhabdovirus components and that it cannot be attributed exclusively to interferon.