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AssigneeInternational AIDS Vaccine Initiative
InventorsJaap Goudsmit, Jerald C. Sadoff, Wayne Koff
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The present invention is directed to a bacterial delivery system for delivering alphavirus replicon DNA into an animal or animal cells with the replicon encoding one or more heterologous genes to be expressed in the animal or the animal cells. The bacteria are invasive bacteria or attenuated invasive bacteria engineered to contain a DNA vector that encodes the alphavirus replicon in a eukaryotic expression cassette. The heterologous gene can encode an antigen, a therapeutic agent, an immunoregulatory agent, an anti-sense RNA, a catalytic RNA, a protein, a peptide, an antibody or an antigen-binding fragment of an antibody. In a preferred embodiment, the heterologous gene encodes one or more antigens useful as a vaccine for HIV. In addition to the bacterial delivery system, the invention provides methods of introducing and expressing the heterologous gene(s) in animal or animal cells and methods of stimulating or inducing an immune response.
A bacterial delivery system which comprises live invasive bacteria containing a DNA comprising a eukaryotic expression cassette operably linked to an alphavirus replicon DNA capable of amplification as RNA in animal cells, wherein the alphavirus replicon DNA comprises at least one nucleic acid control sequence operably linked to a heterologous nucleic acid sequence to control expression of said heterologous gene.
A method for introducing and expressing a gene in an animal which comprises infecting said animal with live invasive bacteria containing a DNA comprising a eukaryotic expression cassette operably linked to an alphavirus replicon DNA capable of amplification as RNA in animal cells, wherein the alphavirus replicon DNA comprises at least one nucleic acid control sequence operably linked to a heterologous nucleic acid sequence to control expression of said heterologous nucleic acid, and thereby obtaining expression of a gene product encoded by said heterologous nucleic acid sequence in said animal.
A method for inducing an immune response in an animal which comprises infecting said animal with live invasive bacteria containing a DNA comprising a eukaryotic expression cassette operably linked to an alphavirus replicon DNA capable of amplification as RNA in animal cells, wherein the alphavirus replicon DNA encodes at least one antigen or antigenic fragment of a protein, and wherein said antigen or said fragment is expressed at a level sufficient to stimulate an immune response to said antigen or said fragment.
A method for introducing and expressing a gene in animal cells comprising
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