Recombinant biologically contained filovirus vaccine

A multivalent vaccine comprising an effective amount of a recombinant filovirus, wherein the genome of the recombinant filovirus contains a first deletion in one or more nucleotides for a polynucleotide sequence for a viral protein corresponding to Ebola virus VP30 which deletion is effective to prevent expression of a functional viral protein corresponding to Ebola virus VP30 upon infection of a cell with the recombinant filovirus, and wherein the genome contains a mutation in a region that is flanked by NP coding sequences and VP35 coding sequences, a mutation in GP/sGP coding sequences, and/or an insertion within 1,000 nucleotides of the first deletion site or at the first deletion site, or a combination thereof, wherein the genome encodes one or more filovirus glycoproteins, wherein the mutation in the region that is flanked by NP coding sequences and VP35 coding sequences comprises an insertion of a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product and optionally also a deletion of one or more nucleotides in the region that flanks the NP coding sequences and VP35 coding sequences, wherein the mutation in the GP/sGP coding sequences comprises an insertion of a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product and optionally also a deletion of one or more nucleotides in the GP/sGP coding sequences, or wherein the insertion that is within 1,000 nucleotides of or at the first deletion site encodes a prophylactic or therapeutic heterologous gene product.

A method of manufacturing recombinant filovirus, comprising:

• providing supernatant from mammalian cells expressing a recombinant filovirus genome that are cultured in serum free medium so as to result in progeny filovirus, wherein the genome of the recombinant filovirus contains a deletion of one or more nucleotides in a polynucleotide sequence for a viral protein corresponding to Ebola virus VP30, and wherein the deletion is effective to prevent expression of a functional viral protein corresponding to Ebola virus VP30 upon infection of a cell with the recombinant filovirus, and wherein the mammalian cells express a viral protein corresponding to Ebola virus VP30;
• contacting the supernatant with a DNase and a virus inactivating agent, thereby providing an inactivated viral preparation;
• and
• concentrating the inactivated viral preparation.