Vaccine
The present invention relates to the field of immunogenic compositions and vaccines, their manufacture, host cells which can be used in their manufacture and the use of such immunogenic compositions and vaccines in medicine. More particularly, it relates to Klebsiella pneumoniae O-antigens, conjugates comprising a K. pneumoniae O-antigen, host cells suitable for their production and immunogenic compositions or vaccines containing at least one Klebsiella pneumoniae O-antigen.
1 . A host cell comprising:
i) first nucleotide sequences, which are integrated into the host cell genome and comprise polysaccharide synthesis genes for producing a Klebsiella pneumoniae O-antigen polysaccharide, the Klebsiella pneumoniae O-antigen polysaccharide comprising a O1v1 O-antigen polysaccharide, a O2a O-antigen polysaccharide, a O2afg O-antigen polysaccharide, or a O3b O-antigen polysaccharide;
ii) a second nucleotide sequence, which is within one or more plasmids and encodes a heterologous oligosaccharyl transferase;
iii) a third nucleotide sequence, which is within the one or more plasmids and encodes a carrier protein, the carrier protein comprising an inserted consensus sequence: the inserted consensus sequence comprising: D/E-X-N-Z-S/T, wherein X and Z is any natural amino acid except proline; and
iv) a fourth nucleotide sequence, which encodes an ABC transporter, the fourth nucleotide sequence comprising Klebsiella pneumoniae wzm gene and Klebsiella pneumoniae wzt gene.
2 . The host cell according to claim 1 , wherein the polysaccharide synthesis genes for producing the Klebsiella pneumoniae O-antigen polysaccharide comprise Klebsiella pneumoniae wbbM gene, Klebsiella pneumoniae glf gene, Klebsiella pneumoniae wbbN gene, and Klebsiella pneumoniae wbbO gene.
3 . The host cell according to claim 1 , wherein the polysaccharide synthesis genes for producing a Klebsiella pneumoniae O-antigen polysaccharide comprise Klebsiella pneumoniae wbbM gene, Klebsiella pneumoniae glf gene, Klebsiella pneumoniae wbbN gene, Klebsiella pneumoniae wbbO gene, Klebsiella pneumoniae gmlA gene, Klebsiella pneumoniae gmlB gene, and Klebsiella pneumoniae gmlC gene.
4 . The host cell according to claim 1 , wherein the polysaccharide synthesis genes for producing a Klebsiella pneumoniae O-antigen polysaccharide comprise Klebsiella pneumoniae wbbM gene, Klebsiella pneumoniae glf gene, Klebsiella pneumoniae wbbN gene, Klebsiella pneumoniae wbbO gene, Klebsiella pneumoniae wbbY gene, and Klebsiella pneumoniae wbbZ gene.
5 . The host cell according to claim 1 , wherein the polysaccharide synthesis genes for producing a Klebsiella pneumoniae O-antigen polysaccharide comprise Klebsiella pneumoniae manC gene, Klebsiella pneumoniae manB gene, Klebsiella pneumoniae wbdD gene, Klebsiella pneumoniae wbdA gene, Klebsiella pneumoniae wbdB gene, and Klebsiella pneumoniae wbdC gene.
6 . A conjugate comprising a carrier protein and a Klebsiella pneumoniae O-antigen polysaccharide: the Klebsiella pneumoniae O-antigen polysaccharide comprising a O1v1 O-antigen polysaccharide, a O2a O-antigen polysaccharide, a O2afg O-antigen polysaccharide, or a O3b O-antigen polysaccharide; the Klebsiella pneumoniae O-antigen polysaccharide being conjugated to the carrier protein, and the carrier protein being a detoxified Exotoxin A of Pseudomonas aeruginosa:
wherein the O1v1 O-antigen polysaccharide has the structure of -(D-galactan II)n-(D-galactan I)m-GlcNAc, as follows:
wherein: (i) the number of repeat units of the D-galactan II (n) is from 5 to 7; (ii) the number of repeat units of the D-galactan I (m) is from 3 to 5; and (iii) the ratio of the D-galactan II to the D-galactan I is from 2:1 to 1:50;
wherein the O2a O-antigen polysaccharide has the structure of -(D-galactan I)p-GlcNAc, as follows:
wherein the number of repeat units of the D-galactan I (p) is from 15 to 30;
wherein the O2afg O-antigen polysaccharide has the structure of -(D-galactan III)r-GlcNAc, as follows:
wherein the number of repeat units of the D-galactan III (r) is from 5 to 15;
wherein the O3b O-antigen polysaccharide has the structure of Me-P-3(Man-α2-Man-α3-Man-α3)s-Man-α3-Man-α3-GlcNAc, as follows:
wherein the number of repeat units of the Man-α2-Man-α3-Man-α3 (s) is from 10 to 20.
7 . The conjugate according to claim 6 , wherein the Klebsiella pneumoniae O-antigen polysaccharide is the O1v1 O-antigen polysaccharide.
8 . The conjugate according to claim 6 , wherein the Klebsiella pneumoniae O-antigen polysaccharide is the O2a O-antigen polysaccharide.
9 . The conjugate according to claim 6 , wherein the Klebsiella pneumoniae O-antigen polysaccharide is the O2afg O-antigen polysaccharide.
10 . The conjugate of claim 6 , wherein the degree of branching of the O2afg O-antigen polysaccharide ranges from 90% to 100%.
11 . The conjugate according to claim 6 , wherein the Klebsiella pneumoniae O-antigen polysaccharide is the O3b O-antigen polysaccharide.
12 . An immunogenic composition comprising: (a) the conjugate of claim 6 and (b)(i) a pharmaceutically acceptable excipient or (b)(ii) a pharmaceutically acceptable carrier.
13 . An immunogenic composition comprising one or more carrier proteins, a Klebsiella pneumoniae O1v1 O-antigen polysaccharide, a Klebsiella pneumoniae O2a O-antigen polysaccharide, a Klebsiella pneumoniae O2afg O-antigen polysaccharide, and a Klebsiella pneumoniae O3b O-antigen polysaccharide, wherein each of the Klebsiella pneumoniae O1v1 antigen polysaccharide, the Klebsiella pneumoniae O2a O-antigen polysaccharide, the Klebsiella pneumoniae O2afg O-antigen polysaccharide, and the Klebsiella pneumoniae O3b O-antigen polysaccharide are conjugated to the one or more carrier proteins.
14 . The immunogenic composition according to claim 13 , wherein the one or more carrier proteins comprise an inserted consensus sequence; the inserted consensus sequence comprising D/E-X-N-Z-S/T, wherein X and Z is any natural amino acid except proline.
15 . A vaccine comprising the immunogenic composition of claim 12 and an adjuvant.
16 . A vaccine comprising the immunogenic composition of claim 13 and an adjuvant.
17 . A vaccine comprising the immunogenic composition of claim 14 and an adjuvant.
18 . A method of inducing an immune response to Klebsiella pneumoniae in a subject, the method comprising administering a therapeutically or prophylactically effective amount of the immunogenic composition of claim 12 to the subject.
19 . A method of inducing an immune response to Klebsiella pneumoniae in a subject, the method comprising administering a therapeutically or prophylactically effective amount of the immunogenic composition of claim 13 to the subject.
20 . A method of inducing an immune response to Klebsiella pneumoniae in a subject, the method comprising administering a therapeutically or prophylactically effective amount of the immunogenic composition of claim 14 to the subject.
21 . The host cell according to claim 1 , wherein the Klebsiella pneumoniae wzm gene comprises a sequence at least 95% identical to SEQ ID NO: 21 or SEQ ID NO: 34 and the Klebsiella pneumoniae wzt gene comprises a sequence at least 95% identical to SEQ ID NO: 22 or SEQ ID NO: 35.
22 . The host cell according to claim 2 , wherein: (a) the Klebsiella pneumoniae wzm gene comprises a sequence at least 95% identical to SEQ ID NO: 21 or SEQ ID NO: 34, (b) the Klebsiella pneumoniae wzt gene comprises a sequence at least 95% identical to SEQ ID NO: 22 or SEQ ID NO: 35, (c) the Klebsiella pneumoniae gene wbbM comprises a sequence at least 95% identical to SEQ ID NO: 23, (d) the Klebsiella pneumoniae gene glf comprises a sequence at least 95% identical to SEQ ID NO: 24, (e) the Klebsiella pneumoniae gene wbbN comprises a sequence at least 95% identical to SEQ ID NO: 25, and (f) the Klebsiella pneumoniae gene wbbO comprises a sequence at least 95% identical to SEQ ID NO: 26.
23 . The host cell according to claim 3 , wherein: (a) the Klebsiella pneumoniae wzm gene comprises a sequence at least 95% identical to SEQ ID NO: 21 or SEQ ID NO: 34, (b) the Klebsiella pneumoniae wzt gene comprises a sequence at least 95% identical to SEQ ID NO: 22 or SEQ ID NO: 35, (c) the Klebsiella pneumoniae wbbM gene comprises a sequence at least 95% identical to SEQ ID NO: 23, (d) the Klebsiella pneumoniae glf gene comprises a sequence at least 95% identical to SEQ ID NO: 24, (e) the Klebsiella pneumoniae wbbN gene comprises a sequence at least 95% identical to SEQ ID NO: 25, (f) the Klebsiella pneumoniae wbbO gene comprises a sequence at least 95% identical to SEQ ID NO: 26, (g) the Klebsiella pneumoniae gmlA gene comprises a sequence at least 95% identical to SEQ ID NO: 27, (h) the Klebsiella pneumoniae gmlB gene comprises a sequence at least 95% identical to SEQ ID NO: 28, and (i) the Klebsiella pneumoniae gmlC gene comprises a sequence at least 95% identical to SEQ ID NO: 29.
24 . The host cell according to claim 4 , wherein: (a) the Klebsiella pneumoniae wzm gene comprises a sequence at least 95% identical to SEQ ID NO: 21 or SEQ ID NO: 34, (b) the Klebsiella pneumoniae wzt gene comprises a sequence at least 95% identical to SEQ ID NO: 22 or SEQ ID NO: 35, (c) the Klebsiella pneumoniae wbbM gene comprises a sequence at least 95% identical to SEQ ID NO: 23, (d) the Klebsiella pneumoniae glf gene comprises a sequence at least 95% identical to SEQ ID NO: 24, (e) the Klebsiella pneumoniae wbbN gene comprises a sequence at least 95% identical to SEQ ID NO: 25, (f) the Klebsiella pneumoniae wbbO gene comprises a sequence at least 95% identical to SEQ ID NO: 26, (g) the Klebsiella pneumoniae wbbY gene comprises a sequence at least 95% identical to SEQ ID NO: 30, and (h) the Klebsiella pneumoniae wbbZ gene comprises a sequence at least 95% identical to SEQ ID NO: 31.
25 . The host cell according to claim 5 , wherein: (a) the Klebsiella pneumoniae wzm gene comprises a sequence at least 95% identical to SEQ ID NO: 21 or SEQ ID NO: 34, (b) the Klebsiella pneumoniae wzt gene comprises a sequence at least 95% identical to SEQ ID NO: 22 or SEQ ID NO: 35, (c) the Klebsiella pneumoniae manC gene comprises a sequence at least 95% identical to SEQ ID NO: 32, (d) the Klebsiella pneumoniae manB gene comprises a sequence at least 95% identical to SEQ ID NO: 33, (e) the Klebsiella pneumoniae wbdD gene comprises a sequence at least 95% identical to SEQ ID NO: 36, (f) the Klebsiella pneumoniae wbdA gene comprises a sequence at least 95% identical to SEQ ID NO: 37, (g) the Klebsiella pneumoniae wbdB gene comprises a sequence at least 95% identical to SEQ ID NO: 38, and (h) the Klebsiella pneumoniae wbdC gene comprises a sequence at least 95% identical to SEQ ID NO: 39.