Replication-competent controlled alpha-herpesvirus vectors and uses therefore
The present disclosure relates to replication-competent controlled herpesviruses whose transient replication in a desired inoculation site region of a subject can be activated by the delivery of an appropriate heat dose to the inoculation site region. In related recombinant viruses, activation requires delivery of a heat dose in the presence in the inoculation site region of an effective concentration of a small-molecule regulator. The viruses are further engineered to be capable of replicating efficiently in the desired inoculation site region but essentially not in nerve ganglia and other nerve cells.
1. A heat-controlled or heat- and small-molecule regulators-controlled replication-competent controlled alpha-herpesvirus whose replication can be transiently activated in nonneural cells infected with said replication-competent controlled alpha-herpesvirus but that cannot be transiently activated in neural cells infected with said replication-competent controlled alpha-herpesvirus, wherein the replication-competent controlled alpha-herpesvirus is a recombinant alpha-herpesvirus derived from a virus of the group consisting of a herpes simplex virus type 1 (HSV-1), a herpes simplex virus type 2 (HSV-2) and a varicella-zoster virus, said replication-competent controlled alpha-herpesvirus comprising:
(a) a first heterologous promoter that is a nucleic acid sequence that acts as a heat shock promoter, the first heterologous promoter controlling the expression of a first replication-essential gene of the replication-competent controlled alpha-herpesvirus, wherein
(1) the first heterologous promoter is functionally linked to the first replication-essential gene of the replication-competent controlled alpha-herpesvirus, or
(2) the first heterologous promoter is functionally linked to a gene for a heterologous, unregulated or small-molecule-regulated transactivator that has been inserted in the genome of the replication-competent controlled alpha-herpesvirus and the first replication-essential gene is functionally linked to a promoter that is responsive to the transactivator, and
(b) a second heterologous promoter that is known to be active in nonneural cells of a mammalian subject to which the replication-competent controlled alpha-herpesvirus is to be administered but is also known to be essentially inactive in neural cells of the mammalian subject, the second heterologous promoter being functionally linked to a second replication-essential gene of the replication-competent controlled alpha-herpesvirus.
2. The replication-competent controlled alpha-herpesvirus according to claim 1 , wherein the second heterologous promoter is active in a cutaneous or subcutaneous region, or a mucosal membrane.
3. The replication-competent controlled alpha-herpesvirus of claim 1 , wherein the replication-competent controlled alpha-herpesvirus is an HSV-1 or HSV-2 and is lacking a functional ICP47 gene.
4. The replication-competent controlled alpha-herpesvirus of claim 1 , further comprising at least one of an expressed gene from another pathogen, an expressed heterologous gene encoding an immune-modulatory polypeptide and an expressed heterologous gene encoding another polypeptide.
5. The replication-competent controlled alpha-herpesvirus of claim 4 , wherein the expressed gene from another pathogen is a gene encoding an influenza virus surface antigen, an internal protein or parts thereof.
6. The replication-competent controlled alpha-herpesvirus of claim 4 , wherein the expressed gene from another pathogen is a gene encoding a human immunodeficiency virus surface antigen, an internal protein or parts thereof.
7. The replication-competent controlled alpha-herpesvirus of claim 1 , wherein the second heterologous promoter is selected from the promoters of a KRT1, a KRT4, a KRT5, a KRT6A, a KRT10, a KRT11, a KRT13, a KRT77, an MLANA and a TYR gene.
8. The replication-competent controlled alpha-herpesvirus according to claim 4 , wherein replication-competent controlled alpha-herpesvirus further comprises a gene encoding a cytokine.
9. The replication-competent controlled alpha-herpesvirus according to claim 4 , wherein replication-competent controlled alpha-herpesvirus further comprises a gene encoding IL-4, IL-2 or GM-CSF.
10. A vaccine composition comprising an effective amount of the replication-competent controlled alpha-herpesvirus of claim 1 and a pharmaceutically acceptable carrier or excipient.
11. A method for vaccinating a subject against herpetic disease caused by an alpha-herpesvirus comprising administering the replication-competent controlled alpha-herpesvirus of claim 1 to a site in the skin of the subject and administering an activating heat dose to the administration site, wherein the herpetic disease is caused by the alpha-herpesvirus from which the replication-competent controlled alpha-herpesvirus was derived.
12. A method for vaccinating a subject against disease caused by another pathogen comprising administering the replication-competent controlled alpha-herpesvirus of claim 1 further comprising an expressed gene from the other pathogen to a site in the skin of the subject and administering an activating heat dose to the administration site.