Polypeptide and immune modulation
The present invention relates to Roseburia flagellin, and/or a polynucleotide sequence encoding said Roseburia flagellin, and/or a vector comprising said polynucleotide sequence, and/or a host cell, including bacteria, comprising said vector, and/or a host cell, including bacteria, comprising said polynucleotide sequence, for use in modulating the inflammation of a tissue or an organ in a subject.
1. A pharmaceutical composition that comprises:
an isolated, recombinant Roseburia flagellin polypeptide in a unit dose comprising a therapeutically effective amount sufficient to reduce inflammation by increasing a level of IL-10 when the unit dose is administered to a subject suffering from a gastrointestinal inflammatory disorder or an autoimmune disorder, relative to an extent inflammation prior to the administering, wherein the isolated, recombinant Roseburia flagellin polypeptide comprises:
a polypeptide sequence having at least 85% identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST, wherein said polypeptide sequence comprises a region that has at least 90% identity to the polypeptide sequence of amino acids 79-117 of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST; and
a preservative or stabilizer, and
wherein the isolated, recombinant Roseburia flagellin polypeptide binds to TLR5.
2. A method of making the pharmaceutical composition according to claim 1 , the method comprising:
expressing the recombinant Roseburia flagellin polypeptide;
purifying the expressed recombinant Roseburia flagellin polypeptide to produce the isolated, recombinant Roseburia flagellin polypeptide; and
admixing the isolated, recombinant Roseburia flagellin polypeptide with the preservative or stabilizer to form the pharmaceutical composition.
3. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide is a FlaA1 polypeptide.
4. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide comprises a Roseburia hominis flagellin polypeptide or a Roseburia intestinalis flagellin polypeptide.
5. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide comprises a polypeptide sequence that has at least 95% sequence identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST, and at least 95% identity to amino acids 79-117 of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST.
6. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide comprises the polypeptide sequence of SEQ ID NO: 2.
7. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide comprises at least 50 contiguous amino acids of the polypeptide sequence of SEQ ID NO: 2 within the sequence having at least 85% identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST.
8. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide comprises at least 100 contiguous amino acids of the polypeptide sequence of SEQ ID NO: 2 within the sequence having at least 85% identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST.
9. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide comprises at least 150 contiguous amino acids of the polypeptide sequence of SEQ ID NO: 2 having at least 85% identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST.
10. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide comprises at least 175 contiguous amino acids of the polypeptide sequence of SEQ ID NO: 2 having at least 85% identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST.
11. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is a solid composition.
12. The pharmaceutical composition of claim 11 , wherein the pharmaceutical composition is in the form of one or more tablets or capsules.
13. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is an enteric formulation.
14. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide is comprised in a fusion protein.
15. The pharmaceutical composition of claim 1 , wherein the isolated recombinant Roseburia flagellin polypeptide has at least 98% identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST, and at least 98% identity to amino acids 79-117 of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST.
16. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide has at least 99% identity to the polypeptide sequence of SEQ ID NO: 2, as determined by a sequence alignment performed using BLAST.
17. The pharmaceutical composition of claim 1 , wherein the isolated, recombinant Roseburia flagellin polypeptide increases a level of IL-10 by at least 5-fold after contacting the isolated, recombinant Roseburia flagellin polypeptide with a bone marrow dendritic cell in an amount of about 100 ng/mL in vitro, relative to a level of IL-10 prior to the contracting.