Pathogenicity of Sendai viruses adapted into polarized MDCK cells.
ABSTRACT Apically and basally released Sendai viruses (SeV) were obtained after infection of polarized Madin-Darby canine kidney (MDCK) cells grown on permeable membrane culture inserts. After 20 passages of adaptation in MDCK cells, we compared their in vivo and in vitro pathogenicity with the parental Mol-strain of SeV. These viruses had comparable in vitro pathogenicity, but the in vivo pathogenicities were varied. The apically released MDCK-adapted virus showed comparable pathogenicity with the parental virus, in contrast with the basally released MDCK-adapted virus, which showed in vivo attenuation.
Full-textDOI: · Available from: Ryoji Yamaguchi, Aug 02, 2015
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ABSTRACT: Many pathogens initiate infection at the mucosal surfaces; therefore, induction of mucosal immune responses is a first level of defense against infection and is the most powerful means of protection. Although intramuscular injection is widely used for vaccination and is effective at inducing circulating antibodies, it is less effective at inducing mucosal antibodies. Here we report a novel recombinant, attenuated Sendai virus vector (GP42-H1) in which the hemagglutinin (HA) gene of influenza A virus was introduced into the Sendai virus genome as an additional gene. Infection of CV-1 cells by GP42-H1 resulted in cell surface expression of the HA protein. Intranasal immunization of mice with 1,000 plaque forming units (pfu) of GP42-H1 induced HA-specific IgG and IgA antibodies in the blood, bronchoalveolar lavage fluid, fecal pellet extracts and saliva. The HA-specific antibody titer induced by GP42-H1 closely resembles the titer induced by sublethal infection by live influenza virus; however, in contrast to infection by influenza virus, immunization with GP42-H1 did not result in disease symptoms or the loss of body weight. In mice that were immunized with GP42-H1 and then challenged with 5LD(50) (1250 pfu) of influenza virus, no significant weight loss was observed and other visual signs of morbidity were not detected. These results demonstrate that the GP42-H1 Sendai virus recombinant is able to confer full protection from lethal infection by influenza virus, supporting the conclusion that it is a safe and effective mucosal vaccine vector.PLoS ONE 04/2011; 6(4):e18780. DOI:10.1371/journal.pone.0018780 · 3.53 Impact Factor
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ABSTRACT: Reverse genetics allows the generation of recombinant viruses entirely from cDNA. One application of this technology is the creation of reporter-expressing viruses, which greatly increase the detail and ease with which these viruses can be studied. However, there are a number of challenges when working with reporter-expressing viruses. Both the reporter protein itself as well as the genetic manipulations within the viral genome required for expression of this reporter can result in altered biological properties of the recombinant virus, and lead to attenuation in vitro and/or in vivo. Further, instability of reporter expression and purging of the genetic information encoding for the reporter from the viral genome can be an issue. Finally, a practical challenge for in vivo studies lies in the attenuation of light signals when traversing tissues. Novel expression strategies and the continued development of brighter, red and far-red shifted reporters and the increased use of bioluminescent reporters for in vivo applications promise to overcome some of these limitations in future. However, a "one size fits all" approach to the design of reporter-expressing viruses has thus far not been possible. Rather, a reporter suited to the intended application must be selected and an appropriate expression strategy and location for the reporter in the viral genome chosen. Still, attenuating effects of the reporter on viral fitness are difficult to predict and have to be carefully assessed with respect to the intended application. Despite these limitations the generation of suitable reporter-expressing viruses will become more common as technology and our understanding of the intricacies of viral gene expression and regulation improves, allowing deeper insight into virus biology both in living cells and in animals.Antiviral research 01/2014; DOI:10.1016/j.antiviral.2014.01.003 · 3.94 Impact Factor