SYNTHETIC OLIGOSACCHARIDES FOR P. AERUGINOSA VACCINE
The present invention provides synthetic Pseudomonas aeruginosa lipooligosaccharide (LOS)-based oligosaccharides and conjugates containing various P. aeruginosa serotype-specific oligosaccharide antigens or various core P. aeruginosa oligosaccharide structures or motifs. The invention further provides P. aeruginosa LOS-based immunogenic and immunoprotective compositions and antibodies derived therefrom for diagnosing, treating, and preventing infections caused by P. aeruginosa.
1 . A synthetic oligosaccharide 1a or 1b
where each of R 1 and R 2 is independently H, a monosaccharide or an oligosaccharide, X is H, a linker group, or a protecting group; L is a linker and Y is a carrier.
2 . The synthetic oligosaccharide of claim 1 , wherein each of R 1 and R 2 is independently selected from the group consisting of independently H, α-Rha-, α-Glc(1-2)-α-Rha-, β-QuiNAc(1-3)-α-Rha-, β-FucNAc(1-3)-α-Rha-, α-Rha[2,3,4-OAc]-, β-OuiNAc(1-3)-α-Rha[2,4-OAc]-, and β-FucNAc(1-3)-α-Rha[2,4, -OAc]-.
3 . The synthetic oligosaccharide of claim 1 , wherein R 1 and R 2 are selected from one of the combinations listed in the following table:
R 1
R 2
α-Rha-
H
α-Glc(1-2)-α-Rha-
H
H
α-Rha-
H
β-QuiNAc(1-3)-α-Rha-
H
β-FucNAc(1-3)-α-Rha-
H
α-Rha[2,3,4-OAc]-
H
β-QuiNAc(1-3)-α-Rha[2,4-OAc]-
H
β-FucNAc(1-3)-α-Rha[2,4,-OAc]-
4 . The synthetic oligosaccharide of claim 1 , where L is an alkylenethiol linker.
5 . The synthetic oligosaccharide of claim 1 , where Y is a carrier selected from the group consisting of a protein, a peptide, a lipid, a polymer, a dendrimers, a virosomes, and a virus-like particle, or a combination thereof.
6 . The synthetic oligosaccharide of claim 5 , where the carrier is a carrier protein.
7 . The synthetic oligosaccharide of claim 6 , where the carrier protein is selected from the group consisting of a bacterial toxoid, a toxin, an exotoxin, and a nontoxic derivative thereof.
8 . The synthetic oligosaccharide of claim 7 , wherein the carrier protein is selected from the group consisting of tetanus toxoid, tetanus toxin Fragment C, diphtheria toxoid, CRM, cholera toxoid, a Staphylococcus aureus exotoxin or toxoid, Escherichia coli heat labile enterotoxin, Pseudomonas aeruginosa exotoxin A, a genetically detoxified variants thereof, a bacterial outer membrane protein, serotype B outer membrane protein complex (OMPC), outer membrane class 3 porin (rPorB), a porin, keyhole limpet hemocyanine (KLH), hepatitis B virus core protein, thyroglobulin, an albumin, ovalbumin; pneumococcal surface protein A (PspA), pneumococcal adhesin protein (PsaA), a purified protein derivative of tuberculin (PPD), a transferrin binding protein, a peptidyl agonist of TLR-5, and a derivative and/or a combinations of the above carriers.
9 . The synthetic oligosaccharide of claim 8 , wherein the carrier protein is selected from the group consisting of CRM 197, Neisseria meningitidis , bovine serum albumin (BSA), human serum albumin (HSA), poly(lysine:glutamic acid), flagellin of motile bacteria, and a derivative and/or a combination thereof.
10 . The synthetic oligosaccharide of claim 8 , wherein the carrier protein is selected from the group consisting of tetanus toxoid, CRM 197, and OMPC.
11 . A pharmaceutical composition comprising a least one oligosaccharide of claim 1 in an effective amount to stimulate an immune response, optionally further comprising a pharmaceutically acceptable carrier.
12 . The pharmaceutical composition of claim 11 , further comprising an adjuvant.
13 . The pharmaceutical composition of claim 11 , wherein the immune response is an antigen-specific immune response.
14 . A composition comprising a synthetic oligosaccharide of claim 1 and a pharmaceutically acceptable vehicle.
15 . The composition of claim 14 , comprising a plurality of different oligosaccharides, where each oligosaccharide is an oligosaccharide of formula 1a or 1b.
16 . The composition of claim 14 , further comprising an adjuvant.
17 . The composition of claim 16 , where the adjuvant is selected from the group consisting of an aluminum salt, RIBI, a toll-like receptor agonist, AS01, AS02, AS03, AS04, AS05, CpG-oligodeoxynucleotide, MF-59, Montanide ISA-51 VG, Montanide ISA-720, Quil A, QS21, synthetic saponins, an immunostimulating complexe, stearyi tyrosine, a virus-like particle, a reconstituted influenza virosomes, a cytokine, mast cell activator compound 48/80, a liposome, a muramyl dipeptide, SAF-1, and a combination thereof.
18 . The composition of claim 16 , comprising an amount of at least one oligosaccharide sufficient to confer immunity against Pseudomonas.
19 . A composition comprising an oligosaccharide of claim 1 as a vaccine.
20 . An antibody preparation against an oligosaccharide according to claim 1 .
21 . The antibody preparation of claim 20 , where the antibody preparation comprises at least one member chosen from a polyclonal antibody, a monoclonal antibody, a mouse monoclonal IgG antibody, a humanized antibody, a chimeric antibody, a single chain antibodies, a fragment thereof, or a combination thereof.
22 . A method of treating a disease associated with P. aeruginosa infection, comprising administering an effective amount for inducing an immune response against Pseudomonas of an oligosaccharide of claim 1 or an antibody thereto.
23 . A method of treating a disease associated with P. aeruginosa infection, comprising administering to a patient in need thereof an oligosaccharide of claim 1 .
24 . The method of claim 22 , wherein the patient is human.
25 . A method for producing an antibody, the method comprising:
(a) administering to a subject an effective amount of at least one oligosaccharide of claim 1 , for producing an antibodies specific for Pseudomonas;
optionally further comprising an adjuvant; and
(b) isolating the antibody from the subject.
26 . A method for producing a monoclonal antibody, the method comprising:
(a) administering to a subject an effective amount of at least one oligosaccharide of claim 1 , for producing an antibody antibodies specific for Moraxella;
(b) isolating the antibody from the subject;
(c) fusing an antibody producing cell from the subject to a myeloma cell; and
(d) harvesting the antibody produced from a fusion subclone.
27 . The method of claim 25 , wherein the subject is a rabbit or a human.
28 . The method of claim 25 , wherein the subject is a rabbit or a human.
29 . An antibody producing cell obtainable by performing steps (a) to (c) of claim 26 .
30 . An antibody obtainable by performing steps (a) to (d) of claim 26 .
31 . A method of diagnosing the presence of Pseudomonas in a sample, comprising contacting the sample with an antibody of claim 20 .