Modified gene vaccines against avian coronaviruses and methods of using the same
The present invention provides both QuilA-loaded chitosan (QAC)-encapsulated NA vaccine compositions and viral vaccine compositions that encode an Infectious Bronchitis Virus (IBV) spike (S) protein, an IBV nucleocapsid (N) protein, or both the S protein and the N protein. Additionally, the present invention provides methods in which the disclosed vaccines are administered to a subject to induce an immune response against IBV or to vaccinate the subject against IBV.
1 . A vaccine composition comprising a polynucleotide that encodes an infectious bronchitis virus (IBV) spike(S) protein, an IBV nucleocapsid (N) protein, or both the S protein and the N protein, wherein the sequence encoding the S protein comprises one of SEQ ID NOs: 22, 24, 26, 28, 30, or 32 or a sequence with at least 90% identity to one of one of SEQ ID NOs: 22, 24, 26, 28, 30, or 32 and, wherein the sequence encoding the N protein comprises one of SEQ ID NOs: 36 or 38 or a sequence with at least 90% identity to one of SEQ ID NOs: 36 or 38.
2 . The vaccine composition of claim 1 , further comprising an adjuvant.
3 . The vaccine composition of claim 2 , wherein the adjuvant comprises disaggregated spherical nanostructures comprising a saponin fraction isolated from an extract of the bark of Quillaja saponaria (Quil A) and chitosan, and wherein the Quil A and chitosan are present at a ratio between 1:15 and 1:100.
4 . The vaccine composition of claim 3 , wherein the chitosan is functionalized by treatment with 5-formyl-2-furan sulfonic acid and sodium borohydride, such that the chitosan surface is negatively charged.
5 . A method of inducing an immune response against infectious bronchitis virus (IBV) in a subject, the method comprising:
(a) administering a first vaccine composition comprising the vaccine composition of claim 1 in an amount effective to induce the immune response against at least one IBV antigen in the subject; and
(b) administering a second vaccine composition comprising a protein, nucleic acid or viral vectored vaccine composition, the second vaccine composition comprising a polynucleotide encoding an IBV S protein, an IBV N protein, or both or comprising an IBV polypeptide selected from an IBV S protein, an IBV N protein or both, wherein administration of the first vaccine composition and the second vaccine composition induces the immune response against IBV in the subject, wherein administration of the second vaccine composition occurs at least two weeks after administration of the first vaccine composition, and wherein both the first vaccine composition and the second vaccine composition are administered by a route selected from intranasal, intramuscular, oral, or in ovo, and wherein the first vaccine composition and the second vaccine composition are administered via the same route or different routes.
6 . The method of claim 5 , wherein the first vaccine composition and the second vaccine composition are administered intranasally.
7 . The method of claim 5 , wherein the subject is an avian subject.
8 . The method of claim 7 , wherein the avian subject is a chicken.
9 . The method of claim 5 , wherein the second vaccine composition comprises a protein comprising an infectious bronchitis virus (IBV) (S) protein, an IBV nucleocapsid (N) protein, or both the S protein and the N protein.
10 . A method comprising administering the vaccine composition of claim 1 to a subject.
11 . The method of claim 10 , wherein the subject is an avian subject.
12 . The method of claim 10 , wherein the vaccine composition is administered intranasally to the subject.
13 . A vaccine composition comprising a viral vector comprising an infectious bronchitis virus (IBV) (S) protein, an IBV nucleocapsid (N) protein, or both the S protein and the N protein, wherein the viral vector further comprises a polynucleotide encoding the S protein, the N protein or both the S protein and the N protein, and wherein the sequence encoding the S protein comprises one of SEQ ID NOs: 22, 24, 26, 28, 30, or 32 or a sequence with at least 90% identity to one of one of SEQ ID NOs: 22, 24, 26, 28, 30, or 32 and, wherein the sequence encoding the N protein comprises one of SEQ ID NOs: 36 or 38 or a sequence with at least 90% identity to one of SEQ ID NOs: 36 or 38.
14 . The vaccine composition of claim 13 , wherein the viral vector is selected from an adeno-associated virus or a poxvirus.
15 . The vaccine composition of claim 13 , further comprising an adjuvant.
16 . The vaccine composition of claim 15 , wherein the adjuvant comprises disaggregated spherical nanostructures comprising a saponin fraction isolated from an extract of the bark of Quillaja saponaria (Quil A) and chitosan, and wherein the (Quil A) and chitosan are present at a ratio between 1:15 and 1:100.
17 . The vaccine composition of claim 16 , wherein the chitosan is functionalized by treatment with 5-formyl-2-furan sulfonic acid and sodium borohydride, such that the chitosan surface is negatively charged.
18 . A method comprising administering the vaccine composition of claim 13 to a subject.
19 . The method of claim 18 , wherein the subject is an avian subject.
20 . The method of claim 18 , wherein the vaccine composition is administered intranasally to the subject.