ANTI COVID-19 THERAPIES USING CD40 LIGAND FUSION PROTEIN
Compositions and methods are presented for prevention and/or treatment of a coronavirus disease wherein the composition comprises a recombinant entity. The recombinant entity comprises a nucleic acid that encodes a extracellular portion of CD40 ligand (CD40L) coupled by a flexible linker to a coronavirus 2 (CoV2) spike protein and/or a CoV2 nucleocapsid protein.
1 . An expression cassette comprising a promoter operably linked to a recombinant nucleic acid that encodes a extracellular portion of CD40 ligand (CD40L) coupled by a flexible linker to a coronavirus 2 (CoV2) spike protein and/or a CoV2 nucleocapsid protein.
2 . The expression cassette of claim 1 , wherein the flexible linker has between 4 and 50 amino acids, and optionally comprises a (GnS)x sequence.
3 . The expression cassette of claim 1 , wherein the recombinant nucleic acid encodes a fusion protein having at least 85% identity to SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11.
4 . A replication defective adenovirus, comprising:
an E1 gene region deletion;
an E2b gene region deletion; and
a nucleic acid encoding a promoter operably coupled to a recombinant nucleic acid that encodes a extracellular portion of CD40 ligand (CD40L) coupled by a flexible linker to a coronavirus 2 (CoV2) spike protein and/or a CoV2 nucleocapsid protein.
5 . The adenovirus of claim 4 , wherein the CoV2 nucleocapsid protein has at least 85% identity to SEQ ID NO:1.
6 . The adenovirus of claim 5 , wherein the CoV2 nucleocapsid protein is fused to an endosomal targeting sequence (N-ETSD) having at least 85% identity to SEQ ID NO:2.
7 . The adenovirus of claim 4 , wherein the CoV2 spike protein has at least 85% identity to SEQ ID NO:4 or SEQ ID NO:5.
8 . The adenovirus of claim 7 , wherein the recombinant nucleic acid encodes a fusion protein having at least 85% identity to any one or more of SEQ ID NOs:6-11.
9 . The adenovirus of claim 4 , wherein the adenovirus further comprises a nucleic acid encoding a trafficking sequence, a co-stimulatory molecule, and/or an immune stimulatory cytokine.
10 . The adenovirus of claim 9 , wherein the co-stimulatory molecule is selected from the group consisting of CD80, CD86, CD30, CD30L, ICOS-L, B7-H3, B7-H4, CD70, OX40L, 4-1BBL, GITR-L, TIM-3, TIM-4, CD48, CD58, TL1A, ICAM-1, and LFA3, and wherein the immune stimulatory cytokine is selected from the group consisting of IL-2, IL-12, IL-15, IL-21, IPS1, and LMP1.
11 . A vaccine composition comprising the adenovirus of claim 4 , wherein the composition is formulated for injection.
12 . A method for inducing immunity against CoV2 in a patient in need thereof, the method comprising administering to the patient the vaccine composition of claim 11 .
13 . A recombinant yeast comprising a nucleic acid encoding a promoter operably linked to a recombinant nucleic acid that encodes a extracellular portion of CD40 ligand (CD40L) coupled by a flexible linker to a coronavirus 2 (CoV2) spike protein and/or a CoV2 nucleocapsid protein.
14 . The recombinant yeast of claim 13 , wherein the CoV2 nucleocapsid protein has at least 85% identity to SEQ ID NO:1.
15 . The recombinant yeast of claim 14 , wherein the CoV2 nucleocapsid protein is fused to an endosomal targeting sequence (N-ETSD) having at least 85% identity to SEQ ID NO:2.
16 . The recombinant yeast of claim 13 , wherein the CoV2 spike protein has at least 85% identity to SEQ ID NO:4 or SEQ ID NO:5.
17 . The recombinant yeast of claim 16 , wherein the recombinant nucleic acid encodes a fusion protein having at least 85% identity to any one or more of SEQ ID NOs:6-11.
18 . The recombinant yeast of claim 13 , wherein the recombinant yeast is Saccharomyces cerevisiae.
19 . A vaccine composition comprising the recombinant yeast of claim 13 , wherein the composition is formulated for injection.
20 . A method for inducing immunity against CoV2 in a patient in need thereof, the method comprising administering to the patient the vaccine composition of claim 19 .