COMPOSITION AND METHODS FOR MODULATING TOLL-LIKE RECEPTOR ACTIVITY

1 . A method for the identification of a compound which modulates the dissociation or formation of a heterodimer formed between Toll-like Receptor 14 and CD14, said method comprising the steps of:

providing first and second cellular samples comprising CD14, Toll-like Receptor 4 and Toll-like Receptor 14,

contacting said first and second samples with a Toll-like Receptor 4 agonist,

contacting said first sample only with a candidate modulator agent under conditions permissive of binding of said agent to at least one of CD14 and Toll-like Receptor 14, and

monitoring the activation status of the Toll-like Receptor 4 receptor complex through a comparison of the level of downstream intracellular signalling between said first and second samples,

wherein a change in Toll-like Receptor 4 signalling between said first sample and said second sample identifies the candidate modulator agent as a modulator of the dissociation or formation of the heterodimer between CD14 and Toll-like Receptor 14.

2 . (canceled)

3 . A method for the identification of a compound which inhibits the binding of a Toll-like Receptor 4 agonist to CD14, said method comprising the steps of:

providing first and second cellular samples comprising CD14,

contacting said first and second samples with a Toll-like Receptor 4 agonist,

contacting said first sample only with a candidate modulator agent under conditions permissive of binding of said agent to CD14, and

monitoring the activation status of the Toll-like Receptor 4 receptor complex through a comparison of the level of downstream intracellular signalling between said first and second samples,

wherein a reduction in Toll-like Receptor 4 signalling between said first sample and said second sample identifies the candidate modulator agent as an inhibitor of the binding of a Toll-like Receptor 4 agonist to CD14.

4 . The method of claim 1 , wherein the TLR4 agonist is lipopolysacchande (LPS).

5 . The method of claim 1 , wherein the level of Toll-like Receptor 4 intracellular signalling is determined by monitoring markers indicative of Toll-like Receptor 4 activity selected from the group consisting of: NF-kappaB activation, and IRF3 protein activation.

6 . A method for the identification of an agent which acts as an antagonist of Toll-like Receptor 4 activation and intracellular signalling, said method comprising the steps of:

providing first and second cellular samples containing Toll-like Receptor 14, Toll-like Receptor 4 and CD 14,

labelling the Toll-like Receptor 14 with a first fluorophore molecule and the CD14 with a second fluorophore molecule,

contacting said first and second samples with a Toll-like Receptor 4 agonist,

contacting said first sample only with a candidate modulator agent under conditions permissive of binding of Toll-like Receptor 4 and/or Toll-like Receptor 14, and

monitoring the interaction of CD14 and TLR14 to determine any dissociation which occurs in the presence of the Toll-like Receptor 4 agonist and the candidate agent by monitoring the fluorescence of the fluorophores,

wherein a decrease in the level of fluorescence is indicative of the candidate modulator agent not being an antagonist of Toll-like Receptor 4 activation and intracellular signalling.

7 . (canceled)

8 . A method for modulating Toll-like Receptor 4 activation and/or intracellular signaling, comprising:

contacting a cellular sample comprising CD14, Toll-like Receptor 4 and Toll-like Receptor 14 with an agent which has one or more of the following properties:

(i) inhibits the dissociation of a heterodimer complex formed between Toll-like Receptor 14 and CD14 when a Toll-like Receptor 4 agonist is bound to at least one of the Toll-like Receptor 4 or the CD14,

(ii) inhibits the formation of a heterodimer complex comprising Toll-like Receptor 14 and CD14,

(iii) inhibits the association of a Toll-like Receptor 4 agonist with CD14,

(iv) inhibits the transfer of a Toll-like Receptor 4 agonist which is bound to CD14 to Toll-like Receptor 14,

(v) inhibits the binding of a Toll-like Receptor 4 activating ligand to CD14, or

(vi) inhibits the transfer of a Toll-like Receptor 4 activating ligand from CD14 to Toll-like Receptor 4, thereby modulating Toll-like Receptor 4 activation and/or intracellular signalling.

9 . The method of claim 8 , wherein the agent is selected from the group consisting of: a protein, a peptide, a peptidomimetic, a small molecule compound, a nucleic acid, a polynucleotide, a polysaccharide, an oligopeptide, a carbohydrate, a lipid, naturally occurring compounds, and an antibody or a fragment thereof.

10 .- 38 . (canceled)

39 . A method for enhancing an immune response mediated by Toll-like Receptor 4, comprising administering to a subject in need thereof an effective amount of an agent which promotes the dissociation of a heterodimer formed between Toll-like Receptor 14 and CD14.

40 . (canceled)

41 . A method for the treatment and/or prophylaxis of a disease condition which is mediated by Toll-like Receptor 4 activation and/or Toll-like Receptor 4 intracellular signalling, the method comprising:

providing an effective amount of an agent which has one or more of the following properties:

(i) inhibits the dissociation of a heterodimer complex comprising Toll-like Receptor 14 and CD14,

(ii) inhibits the formation of a heterodimer complex comprising Toll-like Receptor 14 and CD14,

(iii) inhibits the association of a Toll-like Receptor 4 agonist with CD14,

(iv) inhibits the transfer of a Toll-like Receptor 4 agonist which is bound to CD14 to Toll-like Receptor 14,

(v) inhibits the binding of a Toll-like Receptor 4 activating ligand to CD14, or

(vi) inhibits the transfer of a Toll-like Receptor 4 activating ligand from CD14 to Toll-like Receptor 4, and

administering the same to a subject in need of such treatment.

42 .- 45 . (canceled)

46 . The method of claim 41 , wherein the disease condition which is mediated by Toll-like Receptor 4 activation and/or Toll-like Receptor 4 intracellular signalling is an inflammatory condition, or a neurological condition or neurodegenerative disorder.

47 .- 48 . (canceled)

49 . A method as claimed in claim 46 , wherein the neurodegenerative disorder or neurological condition is chosen from one or more of the group comprising: Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), traumatic brain injury, spinal cord injury, multiple sclerosis, ischemia or ischemia-induced injury, stroke, or a neurodegenerative condition or disorder caused by a bacterial infection.

50 . The method of claim 1 , wherein the candidate modulator agent inhibits the dissociation or formation of the heterodimer between CD14 and Toll-like Receptor 14.

51 . The method of claim 1 , wherein the candidate modulator agent promotes the dissociation or formation of the heterodimer between CD14 and Toll-like Receptor 14.