Vesicular stomatitis virus VSV

Vandenbroeck et al.7 used an ELISA to determine the recovery of immu-noreactive porcine interferon-gamma (IFN-y) from E. coli inclusion bodies. The ELISA used a polyclonal coating antibody with detection by a MAb. The inclusion bodies were solubilized in diluted 6 M guanidine/HCl and IFN subsequently refolded by its removal. The antiviral activity of the interferon was measured with a bioassay using the cytopathic effect (CPE) of vesicular stomatitis virus (VSV) on bovine kidney cells. The results of this study showed that the immu-noreactivity measured by ELISA matched the biological activity measured by bioassay. 

One experimental tool in this direction is provided by some enveloped animal viruses which mature at the cell surface of infected cells (K Sri inen and Renkonen, 1977 Lenard, 1978). Such viruses include influenza virus, Semliki Forest virus (SFV), Sindbis virus, and vesicular stomatitis virus (VSV). They are extremely simple in makeup and hence are very well characterized. They can be tagged with biochemical probes in many different ways. They infect many animal cells in culture, and after infection turn the cells into factories for the production of virus progeny. The protein-synthesizing machinery of the host cell is programmed by the viral RNA to make viral proteins exclusively and these include the viral surface glycoproteins. These are synthesized with signal peptides and inserted into the ER membrane (Katz et ai, 1977 Garoff et... 

The following cell cultures and virus have shown to be suitable MDBK cells (ATCC No. CCL22), or Mouse L cells (NCTC clone 929 ATCC No. CCL I) as the cell culture and vesicular stomatitis virus VSV Indiana strain (ATCC No. VR-158) as the infective agent or human diploid fibroblast FS-71 cells responsive to interferon as the cell culture, and encephalomyocarditis virus (ATCC No. VR-129B) as the infective agent. 

Fig. 38.—Structure of Glycan of Vesicular Stomatitis Virus (VSV) Membrane Glyco-protein.616...

Various types of adherent cells were grown on a coverslip, which was then laid on top of the LAPS chip. Measurements were made for acidification of (1) normal human epidermal keratinocytes stimulated by epidermal growth factor (EGP) or organic chemicals, and of (2) human uterine sarcoma cells as a response to doxorubicin and vincristine (chemotherapeutic drugs). In addition, the inhibition (by ribavirin) of the viral infection of murine fibroblastic L cells by vesicular stomatitis virus (VSV) was investigated by following the acidification rate. A limitation of these studies is the requirement for a low-buffered medium (low bicarbonate content) to achieve maximum sensitivity [847]. 

The assay for interferon involves incubating cells overnight with increasing dilutions of interferon and then challenging the cells with, say, vesicular stomatitis virus (VSV) at 20 p.f.u. per cell. Twenty hours later the culture fluids are harvested and assayed for VSV using a plaque assay ( 14.3.1) on mouse cells. The greatest dilution of interferon which inhibits virus yield by 3.2 fold (0.5 log10) contains 1 unit of interferon (Baron, 1969). 

The oncolytic viruses include adenovirus, measles, reovirus, vesicular stomatitis virus (VSV),HSV,poxvirus, and vaccinia. Specific examples include (1) ONYX-015, which is an adenoviral oncolytic virus, administered to patients with liver metastases of colorectal cancer and pancreatic cancer [29], (2) Reolysin, which is an oncolytic reovirus administered to patients with glioma [30], and (3) MV-CEA, which is an oncolytic measles virus expressing carcinoembryonic antigen, administered to patients with ovarian cancer [31]. Some oncolytic viruses are wild type and are apparently not pathogenic in humans, such as the Newcastle disease virus (NDV), which is an RNA avian paramyxovirus. PV701, a naturally attenuated, replication-competent strain of NDV, has been administered to patients with advanced solid tumors [32], The applicability of oncolytic viruses as a therapy for clinical oncology trials is due to their potential selectivity the ability to kill tumor cells but not normal cells. However, the level of attenuation of viral replication in normal cells is limited for most oncolytic vectors. 

In 1986, venustatriol (2) was isolated from the red algae Laurencia venusta [5]. This compound was shown to possess many of the same structural features as thyrsiferol (1), with the exception of the stereocenters at Cig and C19 as indicated in Fig. (1). At the time of its isolation, venustatriol (2) was reported to display anti-viral activity against vesicular stomatitis virus (VSV) and herpes simplex virus type 1 (HSV-1). The absolute configuration of venustatriol (2) was verified by x-ray crystallography, an experiment that facilitated the assignment of the absolute stereochemistry of thyrsiferol (1). 

Holothurinosides A (75), C (76) and D (77) from Holothuria forskali and the desulfated derivative 78 at a concentration 20 )Xg/ml caused an inhibition of cytopathic effect induced by vesicular stomatitis virus (VSV) in cell culture (20% inhibition of VSV in baby hamster kidney cell line) [68]. 

An in vitro study examined the ability of six components of a a cat s claw bark extract to inhibit two RNA viruses, vesicular stomatitis virus (VSV) and rhinovirus type IB (HRV IB) (Aquino et al., 1989). Three major glycosides and three other quinovic acid glycosides were studied. An inhibitory effect was evident for all six compounds at relatively high concentrations (i.e., the MIC50 approached the concentration that affected host cell morphology and growth). 

Target RNA species that have been reported to be inhibited include HIV (28), vesicular stomatitis virus (VSV) (76), n-myc(109), and a number of normal cellular genes (110-113). However, to demonstrate that RNase H is not involved in effects observed, it is necessary to use antisense drugs that do not form duplexes that are RNase H substrates (e.g., fully modified 2 -oligonucleotides). 

In both procedures, a gene encoding an abundant membrane glycoprotein (G protein) from vesicular stomatitis virus (VSV) Is Introduced Into cultured mammalian cells either by transfection or simply by Infecting the cells with the virus. The treated cells, even those that are not specialized for secretion, rapidly synthesize the VSV G protein on the ER like normal cellular secretory proteins. Use of a mutant encoding a temperature-sensitive VSV G protein allows researchers to turn subsequent protein transport on and off. At the restrictive temperature of 40 °C, newly made VSV G protein Is misfolded and therefore retained within the ER by quality control mechanisms discussed in Chapter 16, whereas at the permissive temperature of 32 C, the accumulated... 

Such direct basolateral-apical sorting has been investigated in cultured Madin-Darby canine kidney (MDCK) cells, a line of cultured polarized epithelial cells (see Figure 6-6). In MDCK cells infected with the influenza virus, progeny viruses bud only from the apical membrane, whereas in cells infected with vesicular stomatitis virus (VSV), progeny viruses bud only from the basolateral membrane. This difference occurs because the HA glycoprotein of influenza... 

There have been numerous other host resistance models used for immunotox-icity testing and these have been discussed and reviewed by Burleson and Burleson (2007). The Candida albicans fungal model (Herzyk et al., 2001) is another important model and may be used instead of or in addition to the Listeria monocytogenes model. Candida-specr c IgG and cytokines may also be quantified. Other bacterial host resistance models include Pseudomonas aeruginosa, Escherichia colt, Staphylococcus aureus, and Klebsiella pneumoniae, parasitic host resistance models including malaria and Trichinella spiralis, and viral host resistance models including encephalomyocarditis (EMC) virus, vesicular stomatitis virus (VSV), and reovirus. 

To study the antiviral activity, Type 1 simplex herpes virus, KOS stock (HSV-Hg) and Indiana vesicular stomatitis virus (VSV) were used and HeLa cells were cultured in Eagle-modified Dulbecco medium (EMDM), supplemented w ith 10% foetal calf serum, on plates with 24 holes. Monolayers of these cells were infected with HSV-1 or VSV at 0.5 ufp/cell or 0.01 ufp/cell, respectively, and later the product to be assayed, pre-dissolved in DMSO, was added in concentrations of 10, 20, 50, 100 and 200 pg/ml. After 48 h incubation for HSV-1 and 24 h for VSV, at 37°C in CO2 atmosphere, the cytopathic CPE effect was measured on a phase-contrasting microscope [85],... 

These cell-agglutinating proteins have been reported active against certain membrane-containing viruses. Concanavalin A (con A, from Canavalia ensiformis) was found to inactivate HSV, vesicular stomatitis virus (VSV), influenza virus and CMV infectivity and also found to interfere with the viral replication [11]. Other examples for these toxins are lentil lectin... 

Apparently related to the problem of host cell protein synthesis inhibition by viruses is the phenomenon of viral interference. In some cases, superinfection of a cell already infected with one virus does not affect the yield of the original virus. In other cases, however, the superinfecting virus completely inhibits translation of the original viral mRRA. Such is the case for superinfection of vesicular stomatitis virus (VSV)-infected cells by poliovirus (32). The kinetics and general properties of the shut-off of VSY protein synthesis appear to be the same as the shut-off of cellular protein synthesis after poliovirus infection... 

The first explanation was based on measurements of the relative affinity of EMC RNA,Vesicular Stomatitis Virus (VSV) mMA, and... 

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