Replication of the linear adenovirus DNA molecule is thought to result from semiconservative synthesis off linear templates, starting from origins at either end of the genome. Recently, however, it has been shown that in cells infected with adenovirus type 5 (Ad5) a significant fraction of the ends of viral DNA molecules become joined head‐to‐tail due at least in part to the formation of covalently closed circles. Circular DNA is not present in virions but joining of the ends of viral DNA is detectable shortly after infection, well before the onset of viral DNA replication. To learn more about the structure and possible function of these circular forms of viral DNA, I have cloned Ad5 circles as plasmids replicating in Escherichia coli. Two plasmids have been analyzed in detail and shown to generate infectious virus with an efficiency comparable with that of virion DNA following transfection into human cells. These results suggest that circles are not totally inert or functionless but that, once formed, they are capable of re‐entering the pool of replicating molecules to generate linear progeny.

A recombinant adenovirus (Ad) expressing glycoprotein B (gB) of herpes simplex virus (HSV) type 1 (AdgB8) was evaluated as a mucosal vaccine candidate. When administered intranasally (inl) to C57B1/6 mice, AdgB8 induced levels of serum anti-HSV gB IgG antibodies similar to those of mice immunized intraperitoneally (ip), which neutralized both HSV-1 and -2. Mice immunized inl with AdgB8 produced secretory IgA specific for HSV gB, but mice immunized ip did not. Splenic anti-HSV cytotoxic T lymphocytes (CTL) were observed after inl and ip immunization; however, there was a time-dependent decrease in the anti-HSV CTL activity from spleens of inl immunized mice. Anti-HSV CTL were also present in the mediastinal lymph nodes after inl but not ip AdgB8 immunization. Furthermore, mice immunized inl with AdgB8 were protected against heterologous inl challenge with HSV-2, and this protection lasted longer than in ip-immunized mice. These results indicate that mucosal immunization with a recombinant adenovirus can induce mucosal and systemic immune responses and provide long-term protection from mucosally or sexually transmitted viruses.

This report describes the design and construction of three between MCMV and MLP was HCMV which was able to recombinant adenoviruses of serotype 5 (Ad5) expressing induce significant amounts of elafin, particularly in human elafin (EL), also called elastase-specific inhibitor. Three A549 cells. When compared in vivo in rat lungs, results promoters were chosen to drive the synthesis of elafin: the were similar; MCMV was the only promoter which induced small (380 bp) human cytomegalovirus promoter (HCMV), significant amounts of elafin as assessed by Northern blot the Ad2 major late promoter (MLP) and the mouse cytoanalysis and ELISA, even with a low dose of virus megalovirus (MCMV) promoter. Human alveolar epithelial (3 x 10 8 p.f.u.). Our data indicate that the MCMV promoter cells (A549), as well as rat and human primary pulmonary is the promoter of choice for the strong induction of fibroblasts were infected with Ad5-HCMV-EL, Ad5-MLPadenovirus-mediated transgenes in the lung, and suggest EL, Ad5-MCMV-EL and with the control Ad5-dl70/3. The its suitability both in rodent experimental models and in MCMV promoter was the most efficient promoter in all cells humans for investigative and therapeutic purposes. studied. MLP was the least efficient promoter. Intermediate

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