Isogenic adenoviruses type 5 expressing or not expressing the E1A gene: efficiency as virus vectors in the vaccination of permissive and non-permissive species.

We have previously constructed a recombinant adenovirus type 5 (Ad5) vector expressing the gD gene of pseudorabies virus (Ad-gD). In this virus the E1A gene was deleted, so that Ad-gD was replication defective. When high doses of this virus were inoculated into target species, it was nevertheless able to elicit a strong and protective immune response. We have restored a functional E1A gene in Ad-gD, by rescuing the E1A transcription unit in an ectopic position, far from the enhancer sequences of E1A. Unlike Ad-gD, this new virus (Ad-gD-E1A) was able to replicate in non-transcomplementing Vero cells. The kinetics of replication of Ad-gD-E1A were similar to that of the wild-type Ad5 when cells were infected at medium or high m.o.i. Nevertheless, the level of virus replication was low when Ad-gD-E1A was plated at low m.o.i. Cotton rats and mice were vaccinated once with 10(9) to 10(5) TCID50 of each of the two viruses. In both species, Ad-gD and Ad-gD-E1A induced similar antibody responses for each dose tested. Both were able to give a high level of protection against the challenge. However, the protective dose of Ad-gD-E1A was at least 250-fold lower in cotton rats, an Ad5-permissive species, than that of Ad-gD. In mice, a species restricted for Ad5 replication, the protective dose of Ad-gD-E1A was still not significantly different from that of Ad-gD. These data indicate that the protective dose of such an adenovirus vector vaccine for an animal species is strictly linked to the capacity of the virus to replicate in this species, at least in the model used. So, in designing an adequate vector one needs to balance the use of low dose vaccines against risks in terms of biosafety. Nevertheless, for diseases in which the protective immune response is mainly based on the antibody response, defective and non-defective adenovirus vectors should demonstrate similar protective doses.