Anti-Pathogen Treatments

Abstract

Chimeric molecules that contain at least one pathogen-detection domain and at least one effector domain, and their methods of use in preventing or treating a pathogen infection in a cell or organism are described. The pathogen-detection domain and effector domain of the chimeric molecules are domains not typically found in nature to be associated together. Agents are also described herein having at least one pathogen-interacting molecular structure and at least one effector-mediating molecular structure, the agent being one that is non-naturally-occurring in a cell. The methods of prevention and treatment described herein are effective for a broad spectrum of pathogens and exhibit little or no toxic side-effects. Assays for the detection of a pathogen, pathogen component, or product produced or induced by a pathogen, are also provided.

1 - 28 . (canceled) 
     
     
         29 . A chimeric molecule having at least one pathogen-detection domain and at least one effector domain, said chimeric molecule being one that is non-naturally-occurring in a cell, wherein said pathogen-detection domain is isolated from an intracellular molecule and said effector domain comprises an apoptosis mediator domain. 
     
     
         30 . The chimeric molecule of  claim 29 , wherein the chimeric molecule further comprises a protein transduction tag. 
     
     
         31 . The chimeric molecule of  claim 30 , wherein the protein transduction tag is selected from the group consisting of: a TAT tag, a PTD-4 tag and an arginine-rich sequence. 
     
     
         32 . An agent comprising the chimeric molecule of  claim 29 . 
     
     
         33 . A method of treating or preventing a pathogen infection in a cell, comprising administering to said cell the chimeric molecule of  claim 29 , whereby in the presence of a pathogen in the cell, said chimeric molecule or agent binds to the pathogen and activates said effector domain, thereby treating or preventing the pathogen infection in said cell. 
     
     
         34 . An assay for the detection of a pathogen infection in a cell, comprising the steps of:
 a) culturing said cell in a suitable culture medium;   b) administering to said cell the chimeric molecule of  claim 29 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.   
     
     
         35 . An assay for the detection of a pathogen infection in an organism, comprising the steps of:
 a) adding a sample obtained from an organism to an uninfected cell;   b) culturing said cell in a suitable culture medium;   c) administering to said cell the chimeric molecule of  claim 29 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.   
     
     
         36 . A chimeric molecule that is non-naturally-occurring in a cell comprising an inducible promoter operatively linked to an effector domain, wherein said inducible promoter is a pathogen-induced product detection domain, and wherein said inducible promoter is selected from the group consisting of: a cytokine-inducible promoter, a double-stranded RNA inducible promoter, an apoptosis-inducible promoter, an unfolded-protein response promoter, an endoplasmic reticulum-associated degradation response promoter, a stress response promoter, and an inflammatory response promoter. 
     
     
         37 . The chimeric molecule of  claim 36 , wherein said inducible promoter is selected the group consisting of: a p53 inducible promoter, a NF-kappa-B-inducible promoter, and a heat shock protein 70 (HSP70). 
     
     
         38 . The chimeric molecule of  claim 36 , wherein said inducible promoter comprises a response element, wherein said response element is selected the group consisting of: an interferon-stimulated response element (ISRE) and an unfolded-protein response element (UPRE). 
     
     
         39 . The chimeric molecule of  claim 36 , wherein said effector domain is selected from the group consisting of: a caspase, a truncated importin α4, an RNase III, a  Salmonella  SpiC and a heat shock protein (Hsp90). 
     
     
         40 . The chimeric molecule of  claim 36 , wherein the chimeric molecule further comprises a protein transduction tag. 
     
     
         41 . The chimeric molecule of  claim 40 , wherein the protein transduction tag is selected from the group consisting of: a TAT tag, a PTD-4 tag and an arginine-rich sequence. 
     
     
         42 . An agent comprising the chimeric molecule of  claim 36 . 
     
     
         43 . A method of treating or preventing a pathogen infection in a cell, comprising administering to said cell the chimeric molecule of  claim 36 , whereby in the presence of a pathogen in the cell, said chimeric molecule or agent binds to the pathogen and activates said effector domain, thereby treating or preventing the pathogen infection in said cell. 
     
     
         44 . An assay for the detection of a pathogen infection in a cell, comprising the steps of:
 a) culturing said cell in a suitable culture medium;   b) administering to said cell the chimeric molecule of  claim 36 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.   
     
     
         45 . An assay for the detection of a pathogen infection in an organism, comprising the steps of:
 a) adding a sample obtained from an organism to an uninfected cell;   b) culturing said cell in a suitable culture medium;   c) administering to said cell the chimeric molecule of  claim 36 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.   
     
     
         46 . A chimeric molecule having at least one pathogen-detection domain and at least one effector domain, said chimeric molecule being one that is non-naturally-occurring in a cell, wherein said pathogen-detection domain is isolated from the group consisting of: a double-stranded RNA binding domain, a lipopolysaccharide binding domain, a cytokine binding domain, an inflammatory response binding domain, and wherein said effector domain is isolated from the group consisting of: an apoptosis mediator domain, a cytokine mediator domain, an inflammatory mediator domain, an endoplasmic reticulum mediator domain and an unfolded protein response mediator domain. 
     
     
         47 . The chimeric molecule of  claim 46 , wherein said pathogen-detection domain is selected from the group consisting of: a bactericidal/permeability-increasing protein (BPI), janus kinase 1 (JAK1), Tyk2, I kappa B kinase gamma (IKK gamma), heat shock factor 1 (HSF1), protein kinase R, E3L protein, and an RNase L 2′,5′-oligoadenylate binding domain. 
     
     
         48 . The chimeric molecule of  claim 46 , wherein said effector domain is selected from the group consisting of: PKR-like endoplasmic reticulum kinase (PERK), RNase L, IKK gamma, heat shock factor 1 (HSF1), Apaf-1, FLICE Activated Death Domain (FADD), caspase-3, caspase-8 and caspase-9. 
     
     
         49 . The chimeric molecule of  claim 46 , wherein the chimeric molecule further comprises a protein transduction tag. 
     
     
         50 . The chimeric molecule of  claim 49 , wherein the protein transduction tag is selected from the group consisting of: a TAT tag, a PTD-4 tag and an arginine-rich sequence. 
     
     
         51 . An agent comprising the chimeric molecule of  claim 46 . 
     
     
         52 . A method of treating or preventing a pathogen infection in a cell, comprising administering to said cell the chimeric molecule of  claim 46 , whereby in the presence of a pathogen in the cell, said chimeric molecule or agent binds to the pathogen and activates said effector domain, thereby treating or preventing the pathogen infection in said cell. 
     
     
         53 . An assay for the detection of a pathogen infection in a cell, comprising the steps of:
 a) culturing said cell in a suitable culture medium;   b) administering to said cell the chimeric molecule of  claim 46 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.   
     
     
         54 . An assay for the detection of a pathogen infection in an organism, comprising the steps of:
 a) adding a sample obtained from an organism to an uninfected cell;   b) culturing said cell in a suitable culture medium;   c) administering to said cell the chimeric molecule of  claim 46 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.   
     
     
         55 . A chimeric molecule having at least one pathogen-detection domain and at least one effector domain, said chimeric molecule being one that is non-naturally-occurring in a cell, wherein said pathogen-detection domain is isolated from a double-stranded RNA binding compound, and said effector domain comprises a caspase recruitment domain. 
     
     
         56 . The chimeric molecule of  claim 55 , wherein the caspase recruitment domain is selected from Apaf-1 and FLICE Activated Death Domain (FADD). 
     
     
         57 . The chimeric molecule of  claim 55 , wherein the double-stranded RNA binding compound is selected from the group consisting of lividomycin or tobramycin. 
     
     
         58 . The chimeric molecule of  claim 55 , wherein the chimeric molecule further comprises a protein transduction tag. 
     
     
         59 . The chimeric molecule of  claim 58 , wherein the protein transduction tag is selected from the group consisting of TAT, PTD-4, and a polyarginine sequence. 
     
     
         60 . An agent comprising the chimeric molecule of  claim 55 . 
     
     
         61 . A method of treating or preventing a pathogen infection in a cell, comprising administering to said cell the chimeric molecule of  claim 55 , whereby in the presence of a pathogen in the cell, said chimeric molecule or agent binds to the pathogen and activates said effector domain, thereby treating or preventing the pathogen infection in said cell. 
     
     
         62 . An assay for the detection of a pathogen infection in a cell, comprising the steps of:
 a) culturing said cell in a suitable culture medium;   b) administering to said cell the chimeric molecule of  claim 55 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.   
     
     
         63 . An assay for the detection of a pathogen infection in an organism, comprising the steps of:
 a) adding a sample obtained from an organism to an uninfected cell;   b) culturing said cell in a suitable culture medium;   c) administering to said cell the chimeric molecule of  claim 55 , whereby in the presence of a pathogen in said cell, said chimeric molecule or agent binds to said pathogen and activates said effector domain; and   c) determining the presence or absence of effector domain activation;   whereby activation of said effector domain indicates the presence of a pathogen infection in said cell.