Coronavirus T Cell Epitopes and Uses Thereof

Abstract

The present invention includes compositions and methods for detecting the presence of: a coronavirus or an immune response to a coronavirus infection including T cells responsive to one or more coronavirus peptides or proteins comprising, consisting of, or consisting essentially of: one or more amino acid sequences selected from SEQ ID NO: 1 to 1126, subsequences, portions, homologues, variants or derivatives; a fusion protein comprising one or more amino acid sequences selected from SEQ ID NO: 1 to 1126; a pool of peptides or proteins selected from the amino acid sequences set forth in SEQ ID NO: 1 to 1126; or a polynucleotide that encodes one or more peptides or proteins, comprising, consisting of, or consisting essentially of an amino acid sequence selected from SEQ ID NO: 1 to 1126, subsequences, portions, homologues, variants or derivatives. The invention further provides vaccines, diagnostics, therapies, and kits, comprising such proteins or peptides.

Description

Claims

1-157. (canceled)

158. A composition comprising monomers or multimers of: one or more peptides or proteins comprising, consisting of, or consisting essentially of: one or more SARS-CoV-2 amino acid sequences selected from SEQ ID NO: 1 to 1126, concatemers, subsequences, portions, homologues, variants or derivatives thereof; a fusion protein comprising one or more amino acid sequences selected from SEQ ID NO: 1 to 1126.

159. The composition of claim 158, wherein the one or more peptides or proteins or the fusion protein comprises, 22, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1125, or more amino acid sequences selected from SEQ ID NO: 1 to 1126, or a subsequence, portion, homologue, variant or derivative thereof.

160. The composition of claim 158, wherein the one or more protein or peptide comprises at least one of: a SARS-CoV-2 T cell epitope, a SARS-CoV-2 CD8+ or CD4+ T cell epitope, the SARS-CoV-2 T cell epitope is not conserved in another coronavirus, the SARS-CoV-2 T cell epitope is conserved in another coronavirus, or the one or more peptides or proteins exclude the amino acid sequences selected from SEQ ID NOS: 245-280 and 804-873.

161. The composition of claim 158, wherein one or more peptides or proteins has a length from about 9-15, 15-20, 20-25, 25-30, 30-40, 40-50, 50-75 or 75-100 amino acids.

162. The composition of claim 158, wherein the one or more peptides or proteins elicits, stimulates, induces, promotes, increases or enhances a T cell response to SARS-CoV-2, or wherein the one or more peptides or proteins that elicits, stimulates, induces, promotes, increases or enhances the T cell response to SARS-CoV-2 is a SARS-CoV-2 spike, nucleoprotein, membrane, replicase polyprotein 1ab, protein 3a, envelope small membrane protein, non-structural protein 3b, protein 7a, protein 9b, non-structural protein 6, or non-structural protein 8a protein or peptide, or a variant, homologue, derivative or subsequence thereof.

163. The composition of claim 158, further comprising formulating the one or more peptides or proteins into an immunogenic formulation with an adjuvant, wherein the adjuvant is selected from the group consisting of adjuvant is selected from the group consisting of alum, aluminum hydroxide, aluminum phosphate, calcium phosphate hydroxide, cytosine-guanosine oligonucleotide (CpG-ODN) sequence, granulocyte macrophage colony stimulating factor (GM-CSF), monophosphoryl lipid A (MPL), poly(I:C), MF59, Quil A, N-acetyl muramyl-L-alanyl-D-isoglutamine (MDP), FIA, montanide, poly (DL-lactide-coglycolide), squalene, virosome, AS03, ASO4, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, STING, CD40L, pathogen-associated molecular patterns (PAMPs), damage-associated molecular pattern molecules (DAMPs), Freund’s complete adjuvant, Freund’s incomplete adjuvant, transforming growth factor (TGF)-beta antibody or antagonists, A2aR antagonists, lipopolysaccharides (LPS), Fas ligand, Trail, lymphotactin, Mannan (M-FP), APG-2, Hsp70 and Hsp90, pattern recognition receptor ligands, TLR3 ligands, TLR4 ligands, TLR5 ligands, TLR⅞ ligands, and TLR9 ligands.

164. The composition of claim 158, wherein the composition further comprises a modulator of an immune response, an innate immune response, or the modulator is Interleukin-6 (IL-6), Interferon-gamma (IFN-g), Transforming growth factor beta (TGF-B), Interleukin-10 (IL-10), or an agonist or antagonist thereof.

165. A method for detecting the presence of: (i) SARS-CoV-2 or (ii) an immune response relevant to SARS-CoV-2 infections, vaccines or therapies, including T cells responsive to one or more SARS-CoV-2 peptides, comprising: providing one or more proteins or peptides for detection of an amount or a relative amount of, and/or the activity of, and/or the state of antigen-specific T-cells; contacting a biological sample suspected of having SARS-CoV-2-specific T-cells to one or more proteins or peptides for detection; and detecting an amount or a relative amount of, and/or the activity of, and/or the state of antigen-specific T-cells in the biological sample, wherein the one or more proteins or peptides for detection comprise one or more amino acid sequences set forth in SEQ ID NO: 1 to 873, or comprise a pool of 22, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1125, or more amino acid sequences set forth in SEQ ID NO: 1 to 1126.

166. A kit for the detection of SARS-CoV-2 or an immune response to SARS-CoV-2 in a subject comprising: one or more T cells that specifically detect the presence of: one or more amino acid sequences selected from SEQ ID NO: 1 to 1126, or a subsequence, portion, homologue, variant or derivative thereof;a fusion protein comprising one or more amino acid sequences selected from SEQ ID NO: 1 to 1126; ora pool of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1125 more peptides selected from the amino acid sequences set forth in SEQ ID NO: 1 to 1126.

167. The kit of claim 166, wherein the one or more amino acid sequences are selected from at least one of: a coronavirus T cell epitope set forth in SEQ ID NO:874 to 1126; wherein the one or more amino acid sequences comprises: one or more amino acid sequences selected from SEQ ID NO: 1 to 873, or a subsequence, portion, homologue, variant or derivative thereof;a fusion protein comprising one or more amino acid sequences selected from SEQ ID NO: 1 to 873;a pool of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 450, 500, 600, 700, 800, or 873 more peptides selected from the amino acid sequences set forth in SEQ ID NO: 1 to 873; orthe fusion protein has a length from about 9-15, 15-20, 20-25, 25-30, 30-40, 40-50, 50-75 or 75-100 amino acidsa coronavirus CD8+ or CD4+ T cell epitope;the T cell epitope is not conserved in another coronavirus; orthe T cell epitope is conserved in another coronavirus.

168. The kit of claim 166, wherein the kit includes instruction for a diagnostic method, a process, a composition, a product, a service or component part thereof for the detection of: (i) coronavirus or (ii) an immune response relevant to coronavirus infections, vaccines or therapies, including T cells responsive to coronavirus.

169. The kit of claim 166, wherein the kit includes reagents for detecting an amount or a relative amount of, and/or the activity of, and/or the state of antigen-specific T-cells in the biological sample comprises measuring one or more of a cytokine or lymphokine secretion assay, T cell proliferation, immunoprecipitation, immunoassay, ELISA, radioimmunoassay, immunofluorescence assay, Western Blot, FACS analysis, a competitive immunoassay, a noncompetitive immunoassay, a homogeneous immunoassay a heterogeneous immunoassay, a bioassay, a reporter assay, a luciferase assay, a microarray, a surface plasmon resonance detector, a florescence resonance energy transfer, immunocytochemistry, or a cell mediated assay, or a cytokine proliferation assay.

170. The kit of claim 166, wherein the kit includes reagents for determining a Human Leukocyte Antigen (HLA) profile of a subject, and selecting peptides that are presented by the HLA profile of the subject for detecting an immune response to coronavirus.

171. A method of stimulating, inducing, promoting, increasing, or enhancing an immune response against SARS-CoV-2 in a subject, comprising: administering to a subject an amount of a protein or peptide or a polynucleotide that expresses the protein or peptide comprising an amino acid sequence of the SARS-CoV-2 spike, nucleoprotein, membrane, replicase polyprotein 1ab, protein 3a, envelope small membrane protein, non-structural protein 3b, protein 7a, protein 9b, non-structural protein 6, or non-structural protein 8a protein or peptide, or a variant, homologue, derivative or subsequence thereof, wherein the protein or peptide comprises at least two peptides selected from the amino acid sequences set forth in SEQ ID NO:1 to 1126 or a subsequence, portion, homologue, variant or derivative thereof, in an amount sufficient to prevent, stimulate, induce, promote, increase, immunize against, or enhance an immune response against SARS-CoV-2 in the subject.

172. The method of claim 171, wherein the immune response provides the subject with protection against SARS-CoV-2 infection or pathology, or one or more physiological conditions, disorders, illnesses, diseases or symptoms caused by or associated with SARS-CoV-2 infection or pathology, reduces SARS-CoV-2 viral titer, increases or stimulates SARS-CoV-2 viral clearance, reduces or inhibits SARS-CoV-2 viral proliferation, reduces or inhibits increases in SARS-CoV-2 viral titer or SARS-CoV-2 viral proliferation, reduces the amount of a SARS-CoV-2 viral protein or the amount of a SARS-CoV-2 viral nucleic acid, or reduces or inhibits synthesis of a SARS-CoV-2 viral protein or a SARS-CoV-2 viral nucleic acid the SARS-CoV-2 infection is an acute infection, or the subject is a mammal or a human, or the immune response is to two or more circulating forms of SARS-CoV-2 or two or more coronaviruses.

173. The method of claim 171, wherein the one or more amino acid sequences are selected from at least one of: the one or more peptides selected from SEQ ID NO: 1 to 1126, or a subsequence, portion, homologue, variant or derivative thereof;a fusion protein comprising one or more amino acid sequences selected from SEQ ID NO: 1 to 1126; ora pool of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1125 more peptides selected from the amino acid sequences set forth in SEQ ID NO: 1 to 1126.the anti-SARS-CoV-2 T cell response is a CD8+, a CD4+ T cell response, or both; orthe T cell epitope is conserved across two or more clinical isolates of SARS-CoV-2.

174. The method of claim 171, wherein the composition further comprising formulating the one or more peptides or proteins into an immunogenic formulation with an adjuvant, wherein the adjuvant is selected from the group consisting of adjuvant is selected from the group consisting of alum, aluminum hydroxide, aluminum phosphate, calcium phosphate hydroxide, cytosine-guanosine oligonucleotide (CpG-ODN) sequence, granulocyte macrophage colony stimulating factor (GM-CSF), monophosphoryl lipid A (MPL), poly(I:C), MF59, Quil A, N-acetyl muramyl-L-alanyl-D-isoglutamine (MDP), FIA, montanide, poly (DL-lactide-coglycolide), squalene, virosome, AS03, ASO4, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-15, IL-17, IL-18, STING, CD40L, pathogen-associated molecular patterns (PAMPs), damage-associated molecular pattern molecules (DAMPs), Freund’s complete adjuvant, Freund’s incomplete adjuvant, transforming growth factor (TGF)-beta antibody or antagonists, A2aR antagonists, lipopolysaccharides (LPS), Fas ligand, Trail, lymphotactin, Mannan (M-FP), APG-2, Hsp70 and Hsp90, pattern recognition receptor ligands, TLR3 ligands, TLR4 ligands, TLR5 ligands, TLR⅞ ligands, and TLR9 ligands, or a modulator of an immune response, an innate immune response, or the modulator is Interleukin-6 (IL-6), Interferon-gamma (IFN-y), Transforming growth factor beta (TGF-β), Interleukin-10 (IL-10), or an agonist or antagonist thereof.

175. A polynucleotide that expresses one or more peptides or proteins, comprising, an amino acid sequence selected from SEQ ID NO: 1 to 1126, or a subsequence, portion, homologue, variant or derivative thereof; a fusion protein comprising one or more amino acid sequences selected from SEQ ID NO: 1 to 1126; ora pool of 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1125 or more peptides comprising, consisting of, or consisting essentially of amino acid sequences selected from SEQ ID NO: 1 to 1126.

176. A vector that comprises the polynucleotide of claim 175.

177. The vector of claim 176, wherein the vector is a viral vector.

178. A host cell that comprises the vector of claim 176.