Toll-like receptor 5 ligands and methods of use
View Patent ↗The invention provides an immunomodulatory flagellin peptide having at least about 10 amino acids of substantially the amino acid sequence GAVQNRFNSAIT (SEQ ID NO:2), or a modification thereof, and having toll-like receptor 5 (TLR5) binding. Methods of inducing an immune response are also provided.
1. A composition comprising,
(a) a flagellin polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS: 13-33; and
(b) a TLR5 polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:6 and SEQ ID NO:8,
wherein said flagellin polypeptide is detectably labeled.
2. An in vitro method of screening for a TLR5 ligand, agonist or antagonist, comprising:
(a) contacting a TLR5 polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:6 and SEQ ID NO:8 with a candidate compound in the presence of a flagellin polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:13-33 wherein binding of said flagellin polypeptide to said TLR5 polypeptide produces a signal;
(b) determining the production of said signal in the presence of and absence of said candidate compound; and
(c) comparing said signal in the presence of said candidate compound with a signal in the absence of said candidate compound, wherein a difference between said signals in the presence and absence of said candidate compound indicates that said compound is a TLR5 ligand, agonist or antagonist.
3. The method of claim 2 , wherein said signal is amount of a cytokine selected from the group consisting of TNFα, IL-1 and IL-6.
4. The method of claim 2 , wherein said signal is NF-κβ activity.
5. An in vitro assay for detecting bacterial contamination in a sample comprising,
(a) contacting said sample with a TLR5 polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:6 and SEQ ID NO:8 under conditions wherein binding of any flagellin in said sample to said TLR5 polypeptide produces a signal;
(b) determining the production of said signal in the presence and absence of said sample; and
(c) comparing said signal in the presence of said sample with a signal in the absence of said sample, wherein an increase in said signal in the presence as compared to absence of said sample indicates that said sample contains flagellin, whereby bacterial contamination is detected in said sample.
6. The method of claim 5 , wherein said sample is food for an animal.
7. The method of claim 5 , wherein said signal is NF-κβ activity.
8. The method of claim 5 , wherein said signal is an amount a cytokine selected from the group consisting of TNFα, IL-1 and IL-6.