Polypeptides and use thereof
An isolated polypeptide comprises an amino acid sequence of SEQ ID No. 1 or 2 or a variant or fragment thereof. The variant may comprise an amino acid sequence that is at least 70% or 95% identical to the amino acid sequence of SEQ ID No. 1 or 2. A fragment thereof may be a peptide comprising at least 12 contiguous amino acids of SEQ ID No. 1 or 2. The polypeptide exhibits toll-like receptor activity. The TLR has been named TLR1 4. TLR receptors recognise a range of ligands and activate a series of signalling pathways that lead to the induction of immune and inflammatory genes.
1 - 69 . (canceled)
70 . A method of treating an immune or inflammatory disorder, comprising: administering a therapeutically effective amount of a polypeptide comprising the amino acid sequence of SEQ ID No. 1 or 2, or a fragment or variant thereof, to a subject in need thereof.
71 . The method of claim 70 wherein the fragment is chosen from one or more of: an antigenic fragment of SEQ ID NO: 1 or 2, the TLR14 ectodomain shown in FIG. 1E , or the cytoplasmic region of TLR14 shown in FIG. 3 .
72 . The method of claim 70 wherein the variant comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID No. 1 or 2.
73 . The method of claim 70 wherein the variant comprises a modification chosen from one or more of: a deletion modification, an insertion modification, or a post translational modification.
74 . The method of claim 70 wherein the polypeptide or fragment or variant thereof is a soluble or a fusion protein.
75 . The method of claim 74 wherein the polypeptide or fragment or variant thereof is capable of inhibiting the activity of TLR14.
76 . An adjuvant or vaccine formulation wherein the adjuvant or vaccine formulation comprises a polypeptide comprising the amino acid sequence of SEQ ID No. 1 or 2, or a fragment or variant thereof.
77 . A soluble or a fusion a polypeptide comprising the amino acid sequence of SEQ ID No. 1 or 2, or a fragment or variant thereof.
78 . The soluble or fusion polypeptide of claim 77 wherein the fragment is chosen from one or more of: an antigenic fragment of SEQ ID No: 1 or 2, the TLR14 ectodomain shown in FIG. 1E or the cytoplasmic region of TLR14 shown in FIG. 3 .
79 . The soluble or fusion polypeptide of claim 77 wherein the variant comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID No. 1 or 2.
80 . The soluble or fusion polypeptide of claim 77 wherein the fragment or variant thereof is capable of inhibiting the activity of TLR14.
81 . A method for identifying a compound which modulates Toll-like receptor activity comprising the steps of :
contacting a polypeptide comprising an amino acid sequence of SEQ ID No. 1 or 2 or variant or fragment thereof with a test sample comprising one or more compounds;
monitoring for markers of Toll-like receptor activity; and
identifying the one or more compounds which modulate Toll-like receptor activity.
82 . The method of claim 81 wherein the fragment is chosen from one or more of: an antigenic fragment of SEQ ID NO: 1 or 2, the TLR14 ectodomain shown in FIG. 1E , or the cytoplasmic region of TLR14 shown in FIG. 3 .
83 . The method of claim 81 wherein the variant comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID No. 1 or 2.
84 . The method of claim 81 wherein the markers of Toll-like receptor activity comprise any one, two, three or more of:
(i) NFkappaB activation
(ii) NFkappaB protein or polynucleotide encoding the same
(iii) IRF3 protein or polynucleotide encoding the same
(iv) p38 protein or polynucleotide encoding the same
(v) IKKs protein or polynucleotide encoding the same
(vi) RANTES protein or polynucleotide encoding the same
(vii) TLR4 protein or polynucleotide encoding the same or
(viii) any pro-inflammatory or inhibitory cytokine.
85 . The method of claim 81 wherein a compound which activates or inhibits TLR activity is identified by determining the amount, expression, activity or phosphorylation relative to the test sample of a least one or more of:
(i) NFkappaB activation
(ii) NFkappaB protein or polynucleotide encoding the same
(iii) IRF3 protein or polynucleotide encoding the same
(iv) p38 protein or polynucleotide encoding the same
(v) IKKs protein or polynucleotide encoding the same
(vi) RANTES protein or polynucleotide encoding the same
(vii) TLR4 protein or polynucleotide encoding the same or (viii) any pro-inflammatory or inhibitory cytokine.
86 . A method of screening for a compound capable of inhibiting or promoting NF-κB activation comprising the steps of:
providing a host cell comprising a nucleic acid encoding a protein comprising the amino acid sequence of SEQ ID No. 1 or 2, or a variant or fragment thereof and a component that provides a detectable signal associated with activation of NF-κB;
culturing the host cell under conditions providing the expression of the nucleic acid in the host cell;
contacting the host cell with one or more compounds for screening;
measuring the detectable signal; and
isolating or identifying the one or more compounds as an activator compound or an inhibitor compound.
87 . A method of preparing a pharmaceutical composition comprising the steps as claimed in claim 86 including the step of:
optimising the isolated or identified compound as a pharmaceutical compound.
88 . The method of claim 86 wherein the fragment is chosen from one or more of:
an antigenic fragment of SEQ ID NO: 1 or 2, the TLR14 ectodomain shown in FIG. 1E , or the cytoplasmic region of TLR14 shown in FIG. 3 .
89 . The method of claim 86 wherein the variant comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID No. 1 or 2.
90 . The method of claim 86 wherein the protein is a recombinant protein comprising an amino acid sequence of SEQ ID No. 1 or 2.
91 . An isolated nucleic acid comprising a nucleotide sequence of SEQ ID No: 3 or SEQ ID No: 4, or a variant or fragment thereof, or a nucleotide sequence complementary thereto.
92 . A recombinant vector comprising the nucleic acid according to claim 91 .
93 . A host cell comprising the nucleic acid according to claim 91 .
94 . A method of producing a polypeptide comprising culturing the host cell of claim 93 under conditions where the polypeptide is expressed.
95 . An antibody or fragment thereof that specifically binds to an epitope of a polypeptide comprising the amino acid sequence of SEQ ID No. 1 or 2, or a fragment or variant thereof.
96 . A method of treating an immune or inflammatory disorder, comprising: administering a therapeutically effective amount of an antibody or fragment thereof that specifically binds to an epitope of a polypeptide comprising the amino acid sequence of SEQ ID No. 1 or 2, or a fragment or variant thereof, to a subject in need thereof.