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We determined the capacity of vaccinia virus recombinants expressing individual rotavirus genes to induce virus-specific cytotoxic T lymphocytes (CTLs) in mice. Mice were orally inoculated with vaccinia virus recombinants containing genes which encode rotavirus outer capsid proteins vp4 or vp7, single-shelled virus proteins vp1, vp2, or vp6, or rotavirus nonstructural proteins NS53, NS35, NS28, or NS26/NS12. We found that (i) the greatest frequencies of virus-specific CTLs were induced by vaccinia virus recombinants expressing vp7, (ii) transport of vp7 beyond the endoplasmic reticulum was not necessary for induction of CTLs, (iii) recombinants expressing vp7 induced CTLs which reacted with different rotavirus serotypes, and (iv) CTLs were induced among both intestinal and nonintestinal lymphocytes after oral inoculation. These findings may be relevant to vaccine strategies which utilize vectors expressing individual rotavirus genes.

The virological requirements for the recognition of infected target cells by cytolytic T lymphocytes (CTL), using reovirus, a nonenveloped, icosahedral virus has been investigated. Using mouse L cells infected at the nonpermissive temperature with ts (temperature-sensitive) mutants of reovirus in complementation groups C and G, it has been shown that the production of complete viral particles is not necessary for efficient lysis of infected cells by CTL. In addition, adsorption of purified viral particles and viral top component (TC), empty capsids lacking genome ds-RNA, to L cells just prior to use in cytolytic T cell assays is sufficient to produce target cells capable of being lysed, though target production is less efficient than with L cells infected with reovirus. Membrane fluorescence analysis of cells infected with reovirus ts mutants at the nonpermissive temperature and with adsorbed viral particles revealed the presence of the viral sigma 1 protein on the cell surface. For adsorbed particles, the degree of membrane fluorescence paralleled the capacity of CTL to lyse target cells. It is concluded that cells infected with icosahedral, nonenveloped viruses, like cells infected with enveloped viruses, express viral antigens on the cell surface even in the absence of the production of complete viral particles; adsorbed viral particles can be incorporated into the cell membrane in a manner sufficient for recognition and lysis by CTL, in the absence of actual infection of the cells.

As measured by in vitro, 51Cr-release assays, the expression on plasma membranes of two, immediate-early, vaccinia virus-specified cell-surface antigens, with mol wt of 25K-27K and 16K-17K, could be directly correlated with the susceptibility of target cells to lysis by vaccinia virus-specific cytotoxic T cells.