Recovery of recombinant human parainfluenza virus type 1 (HPIV1) from cDNA and use of recombinant HPIV1 in immunogenic compositions and as vectors to elicit immune responses against PIV and other human pathogens

An infectious, self-replicating, recombinant human parainfluenza virus type 1 (HPIV1) that is a complete virus or a sub-viral particle comprising a PIV major nucleocapsid (N) protein, a PIV nucleocapsid phosphoprotein (P), a PIV large polymerase protein (L), and a partial or complete, recombinant HPIV 1 genome or antigenome, said recombinant HPIV1 genome or antigenome including at least one mutation selected from the group consisting of:

• a mutation in the L protein Y942 to an amino acid selected from the group consisting of F, C, N, D, W, S, Q, G, T, V, M, L and A;
• a mutation in the L protein L992 to an amino acid selected from the group consisting of H, I, M, W, A, C, K, N, V and Y;
• a mutation in the L protein of E1711 to another amino acid;
• a mutation in the HN protein T553 to another amino acid;
• a mutation in the C protein R84 to another amino acid;
• a mutation in the C protein R84 to another amino acid and mutation of the P protein E119 to another amino acid;
• a mutation in the P protein E119 to another amino acid;
• a deletion of codons 10-15 of the C protein and codons 13-19 of the P protein;
• a deletion of codons 10-11 of the C protein and codons 13-15 of the P protein;
• a deletion of codons 12-13 of the C protein and codon 15-17 of the P protein;
• a deletion of codons 14-15 of the C protein and codons 17-19 of the P protein;
• and
• a deletion of codons 168-170 of the C protein and codons 172-173 of the P protein.

An immunogenic composition comprising an immunogenically effective amount of an infectious, self-replicating, recombinant human parainfluenza virus type 1 (HPIV 1) in a pharmaceutically acceptable carrier, said HPIV 1 comprising a recombinant HPIV 1 genome or antigenome, a PIV N protein, a PIV P protein, and a PIV L protein, said recombinant HPIV1 genome or antigenome including at least one mutation selected from the group consisting of:

• a mutation in the L protein Y942 to an amino acid selected from the group consisting Of F, C, N, D, W, S, Q, G, 1, V, M, L and A;
• a mutation in the L protein L992 to an amino acid selected from the group consisting of H, I, M, W, A, C, K, N, V and Y;
• a mutation in the L protein of E1711 to another amino acid;
• a mutation in the HN protein T553 to another amino acid;
• a mutation in the C protein R84 to another amino acid;
• a mutation in the C protein R84 to another amino acid and mutation of the P protein E119 to another amino acid;
• a mutation in the P protein E119 to another amino acid;
• a deletion of codons 10-15 of the C protein and codons 13-19 of the P protein;
• a deletion of codons 10-11 of the C protein and codons 13-15 of the P protein;
• a deletion of codons 12-13 of the C protein and codon 15-17 of the P protein;
• a deletion of codons 14-15 of the C protein and codons 17-19 of the P protein;
• and
• a deletion of codons 168-170 of the C protein and codons 172-173 of the P protein.

An infectious, self-replicating, recombinant human parainfluenza virus type 1 (HPIV 1) comprising a PIV major nucleocapsid (N) protein, a PIV nucleocapsid phosphoprotein (P), a PIV large polymerase protein (L), and a partial or complete recombinant HPIV 1 genome or antigenome, said recombinant HPIV 1 genome or antigenome including at least one mutation selected from the group consisting of:

• a mutation in the L protein Y942 to an amino acid selected from the group consisting of F, C, N, D, W, S, Q, G, T, V, M, L and A;
• a mutation in the L protein L992 to an amino acid selected from the group consisting of H, I, M, W, A, C, K, N, V and Y;
• a mutation in the L protein of E1711 to another amino acid
• a mutation in the HN protein T553 to A;
• a mutation in the C protein R84 to G;
• a mutation in the C protein R84 to G and mutation of the P protein E119 to G;
• a mutation in the P protein E119 to G;
• a deletion of codons 10-15 of the C protein and codons 13-19 of the P protein that creates an alanine residue in the P protein at the deletion site;
• a deletion of codons 10-11 of the C protein and codons 13-15 of the P protein that creates a glutamic acid residue in the P protein at the deletion site;
• a deletion of codons 12-13 of the C protein and codon 15-17 of the P protein that creates a glutamic acid residue in the P protein at the deletion site that creates a glutamic acid residue in the P protein at the deletion site;
• a deletion of codons 14-15 of the C protein and codons 17-19 of the P protein that creates an alanine residue in the P protein at the deletion site;
• and
• a deletion of codons 168-170 of the C protein and codons 172-173 of the P protein that creates a serine residue in the C protein at the deletion site.

An infectious, self-replicating, recombinant human parainfluenza virus type 1 (HPIV 1) that is a complete virus or a sub-viral particle, comprising a PIV major nucleocapsid (N) protein, a PIV nucleocapsid phosphoprotein (P), a PIV large polymerase protein (L), and a partial or complete, recombinant HPIV 1 genome or antigenome, wherein said HPIV 1 genome or antigenome is combined with one or more heterologous gene(s) or genome segment(s) encoding one or more antigenic determinant(s) of one or more heterologous pathogen(s) to form a chimeric HPIV 1 genome or antigenome, and wherein the chimeric HPIV 1 genome or antigenome is modified by introduction of a further attenuating mutation that is selected from the group consisting of:

• an amino acid substitution of Glu 1711 in the HPIV 1 L protein;
• a combination of mutations selected from
• (i) F170SMPIV 1/Y942H/L992Fcp45, and
• (ii) F170SMPIV 1/L992F/L1558Icp45;
• one or any combination of mutation(s) in the PIV L protein selected from Y942W, Y942S, Y942Q, Y942T, Y942G, Y942A, Y942V, Y942M, Y942T, Y942L, L992K, L992A, L992Y, L992N, L992V, and L992C;
• incorporation of one or more attenuating host range mutation(s) selected from (i) a mutation at codon 119 of the HPIV 1 P open reading frame (ORF) and corresponding mutation at codon 84 of the HPIV 1 C ORF and (ii) a mutation at codon 553 of the HPIV 1 HN ORF;
• incorporation of one or more attenuating host range mutation(s) specifying one or more amino acid change(s) selected from (i) E119G in HPIV 1 P, (ii) R84G in HPIV 1, and (iii) T553A in HPIV 1 HN;
• and
• one or more partial deletions corresponding to (i) codons 10-11 of the C ORF (ii) codons 12-13 of the C ORF (iii) codons 14-15 of the C ORF, (iv) codons 10-15 of the C ORF, and/or (v) codons 168-170 of the C ORF.