RECOMBINANT VECTORS ENCODING CHIMERIC CORONAVIRUS SPIKE PROTEINS AND USE THEREOF
The present invention provides recombinant vectors encoding a chimeric coronavirus spike protein. The present invention further provides new immunogenic compositions and vaccines comprising these recombinant vectors. Methods of administering these immunogenic compositions and vaccines to animal subjects, including humans, felines, and avians, to protect them against coronaviruses also are included. Methods of making the immunogenic compositions and vaccines alone or in combinations with other protective agents are provided too.
1 . A recombinant vector encoding a chimeric coronavirus spike protein that comprises a spike protein originating from a coronavirus, and a transmembrane domain (TMD) and a C-terminal domain (CTD) of a surface glycoprotein originating from a budding virus that buds from a host cell's plasma membrane (BV pm ), in place of a TMD and a CTD of the coronavirus spike protein; wherein when the recombinant vector is a recombinant BV pm , the TMD and the CTD of the surface glycoprotein originate from a virus species that is different from that of the recombinant BV pm , and wherein the coronavirus spike protein originates from a coronavirus selected from the group consisting of an infectious bronchitis virus (IBV).
2 . (canceled)
3 . The recombinant vector of claim 1 , wherein the surface glycoprotein originates from a BV pm species that is selected from the group consisting of the glycoprotein (G protein) of a vesicular stomatitis virus (VSV).
4 . (canceled)
5 . The recombinant vector of claim 1 , wherein the chimeric coronavirus spike protein comprises an inactivated furin cleavage site.
6 . The recombinant vector claim 1 , wherein the chimeric coronavirus spike protein comprises a central helix that is further stabilized in a prefusion state due to two consecutive amino acid residues at the beginning of the central helix being replaced by a pair of proline residues (2P).
7 .- 14 . (canceled)
15 . The recombinant vector of claim 1 , wherein the IBV is a member of a serotype selected from the group consisting of a Massachusetts serotype, a 4/91serotype, and a QX serotype.
16 . The recombinant vector of claim 15 , wherein the IBV is an IBV-Ma5 strain.
17 . The recombinant vector of claim 15 , wherein the chimeric coronavirus spike protein comprises 90% or greater identity with amino acid residues 19 to 1091 of the amino acid sequence of SEQ ID NO: 4, over the same range of amino acid residues; and wherein said chimeric coronavirus spike protein comprises an inactivated furin cleavage site.
18 . The recombinant vector of claim 15 , wherein the chimeric coronavirus spike protein comprises 90% or greater identity with amino acid residues 19 to 1091 of the amino acid sequence of SEQ ID NO: 6, over the same range of amino acid residues; wherein said chimeric coronavirus spike protein comprises an inactivated furin cleavage site; and wherein the alanine (A) residue at position 859 and the isoleucine (I) residue at position 860 of SEQ ID NO: 6 are replaced by a pair of proline residues (2P).
19 . The recombinant vector of claim 17 , wherein the chimeric coronavirus spike protein further comprises 90% or greater identity with amino acid residues 1092 to 1140 of the amino acid sequence of SEQ ID NOs: 4 or 6, over the same range of amino acid residues.
20 . The recombinant vector of claim 1 , that is a recombinant expression vector selected from the group consisting of a recombinant viral vector and a DNA expression plasmid.
21 . The recombinant expression vector of claim 20 , that is a recombinant viral vector selected from the group consisting of a recombinant herpesvirus of turkeys (HVT), a recombinant attenuated Marek's disease virus 1 (MDV1), a recombinant Marek's disease virus 2 (MDV2), a recombinant MV, a recombinant NDV, and an alphavirus RNA replicon particle (RP).
22 . The recombinant viral vector of claim 21 , that is a HVT.
23 . The recombinant viral vector of claim 21 , wherein the alphavirus RNA replicon particle is a Venezuelan Equine Encephalitis virus (VEEV) replicon particle.
24 . The recombinant expression vector of claim 20 , that is a DNA expression plasmid.
25 . The recombinant expression vector of claim 24 , that encodes an RNA replicon, wherein the RNA replicon is a VEEV RNA replicon.
26 .- 27 . (canceled)
28 . An immunogenic composition comprising the recombinant vector of claim 1 , the recombinant viral vector selected from the group consisting of a recombinant herpesvirus of turkeys (HVT), a recombinant attenuated Marek's disease virus 1 (MDV1), a recombinant Marek's disease virus 2 (MDV2), a recombinant MV, a recombinant NDV, and an alphavirus RNA replicon particle (RP), a DNA expression plasmid, or a synthetic mRNA, and a pharmaceutically acceptable carrier.
29 .- 33 . (canceled)
34 . A vaccine to aid in the protection of an avian from infectious bronchitis due to an infection of IBV in the avian comprising the recombinant vector of claim 1 , the recombinant viral vector selected from the group consisting of a recombinant herpesvirus of turkeys (HVT), a recombinant attenuated Marek's disease virus 1 (MDV1), a recombinant Marek's disease virus 2 (MDV2), a recombinant MV, a recombinant NDV, and an alphavirus RNA replicon particle (RP), a DNA expression plasmid, or a synthetic mRNA, and a pharmaceutically acceptable carrier.
35 . The vaccine of claim 34 , which further comprises at least one non-IBV antigen for eliciting protective immunity to a non-IBV avian pathogen.
36 . The vaccine of claim 34 , that comprises an adjuvant.
37 . The vaccine of claim 34 , that is a non-adjuvanted vaccine.
38 .- 46 . (canceled)
47 . A chimeric coronavirus spike protein that comprises a spike protein originating from an IBV, and a TMD and a CTD of a surface glycoprotein originating from a vesicular stomatitis virus, in place of a TMD and a CTD of the IBV spike protein.
48 . The chimeric coronavirus spike protein of claim 47 , wherein the chimeric coronavirus spike protein comprises 90% or greater identity with amino acid residues 19 to 1091 of the amino acid sequence of SEQ ID NO: 4, over the same range of amino acid residues; and wherein said chimeric coronavirus spike protein comprises an inactivated furin cleavage site.
49 . The chimeric coronavirus spike protein of claim 48 , wherein the chimeric coronavirus spike protein comprises 90% or greater identity with amino acid residues 19 to 1091 of the amino acid sequence of SEQ ID NO: 6, over the same range of amino acid residues; wherein said chimeric coronavirus spike protein comprises an inactivated furin cleavage site; and wherein the alanine (A) residue at position 859 and the isoleucine (I) residue at position 860 of SEQ ID NO: 6 are replaced by a pair of proline residues (2P).
50 . The chimeric coronavirus spike protein of claim 48 , wherein the chimeric coronavirus spike protein further comprises 90% or greater identity with amino acid residues 1092 to 1140 of the amino acid sequence of SEQ ID NOs: 4 or 6, over the same range of amino acid residues.
51 . A nucleic acid encoding the chimeric coronavirus spike protein claim 47 .
52 .- 57 . (canceled)