Methods of determining efficacy of treatments of inflammatory diseases of the bowel

Abstract

Novel methods of determining efficacy of a treatment of inflammatory diseases of the bowel in mammals are provided. The methods are of use in screening and determining the efficacy of treatments of inflammatory diseases of the bowel, and for determining the efficacy response of individual sufferers of inflammatory diseases of the bowel to a given regime. Kits for carrying out the method are also provided.

What is claimed is:  
     
         1 . A method of determining the efficacy of a treatment of inflammatory diseases of the bowel in mammals comprising the steps of: 
 a) measuring the level of at least one anti-inflammatory cytokine and at least one pro-inflammatory cytokine in a biological sample from a mammalian subject;    b) determining the ratio of the at least one anti-inflammatory cytokine to the at least one pro-inflammatory cytokine;    c) administering said treatment;    d) measuring the level of the at least one anti-inflammatory cytokine and the at least one pro-inflammatory cytokine in a biological sample from said mammalian subject at a time following administration of said treatment;    e) determining the ratio of the at least one anti-inflammatory cytokine to the at least one, pro-inflammatory cytokine following administration of said treatment;    wherein an increase in the ratio of anti-inflammatory cytokine to pro-inflammatory cytokine following the administration of said treatment is indicative of an inhibitor of inflammatory diseases of the bowel, and no change or a decrease in the ratio of anti-inflammatory cytokine to pro-inflammatory cytokine following the administration of said treatment is indicative said treatment not being an inhibitor of inflammatory diseases of the bowel.    
     
     
         2 . The method according to  claim 1  wherein the anti-inflammatory cytokine is selected from the group comprising interleukin-10, transforming growth factor-β, interleukin-4, interleukin-5, interleukin-13, and mixtures thereof.  
     
     
         3 . The method according to  claim 2  wherein the anti-inflammatory cytokine is selected from the group comprising interleukin-10, transforming growth factors, and mixtures thereof.  
     
     
         4 . The method according to  claim 1  wherein the pro-inflammatory cytokine comprises interleukin-12, tumour necrosis factor-α, interferon-γ, interleukin-2, and mixtures thereof.  
     
     
         5 . The method according to  claim 4  wherein the pro-inflammatory cytokine comprises interleukin-12, tumour necrosis factor-α, interferon-γ, and mixtures thereof.  
     
     
         6 . The method according to  claim 1 , wherein said ratio of anti-inflammatory cytokine to pro-inflammatory cytokine is the ratio interleukin-10/interleukin-12.  
     
     
         7 . The method according to  claim 1 , wherein said ratio of anti-inflammatory cytokine to pro-inflammatory cytokine is the ratio transforming growth factor-β/interleukin-12.  
     
     
         8 . The method according to  claim 1 , wherein said ratio of anti-inflammatory cytokine to pro-inflammatory cytokine is the ratio interleukin-10/interferon-γ.  
     
     
         9 . The method according to  claim 1  wherein said biological sample comprises urine, plasma, serum, saliva, tissue biopsies, cerebrospinal fluid, peripheral blood mononuclear cells with in vitro stimulation, peripheral blood mononuclear cells without in vitro stimulation, gut lymphoid tissues with in vitro stimulation, gut lymphoid tissues without in vitro stimulation, gut lavage fluids, and mixtures thereof.  
     
     
         10 . The method according to  claim 9  wherein said biological sample comprises serum, peripheral blood mononuclear cells with in vitro stimulation, peripheral blood mononuclear cells without in vitro stimulation, and mixtures thereof.  
     
     
         11 . The method according to  claim 10  wherein said biological sample comprises peripheral blood mononuclear cells with in vitro stimulation, peripheral blood mononuclear cells without in vitro stimulation, and mixtures thereof.  
     
     
         12 . The method according to  claim 9  wherein said in vitro stimulation comprises a mitogen, probiotic, anti-CD3 molecule, and mixtures thereof.  
     
     
         13 . The method according to  claim 10 , wherein said in vitro stimulation comprises a mitogen.  
     
     
         14 . The method according to  claim 13  wherein said mitogen comprises a lipopolysaccharide, lectin, superantigen, and mixture thereof.  
     
     
         15 . The method according to  claim 14 , wherein said lectin comprises concanavalin A, phytohemagglutinin, pokeweed mitogen, and mixtures thereof.  
     
     
         16 . The method according to  claim 1  wherein the means for measuring the levels of said at least one anti-inflammatory cytokine in said biological sample comprises ELISAs, radioimmunoassays, multiplexed ELISAs on microarray platforms, multiplexed ELISAs using coded microspheres coupled with a flow cytometer detection systems, bioassays, Western blots, chromatograph-based separation systems, RT-PCR, competitive reverse transcription PCR, Northern blots, gene arrays, direct measurement of m-RNA, and mixtures thereof.  
     
     
         17 . The method according to  claim 16  wherein the means for measuring the levels of anti-inflammatory cytokines in said biological sample comprises ELISAs, RIAs, multiplexed ELISAs using coded microspheres coupled with a flow cytometer detection systems, and mixtures thereof.  
     
     
         18 . The method according to  claim 17  wherein the means for measuring the levels of said at least one anti-inflammatory cytokine in said biological sample comprises multiplexed ELISAs using coded microspheres coupled with a flow cytometer detection systems.  
     
     
         19 . The method according to  claim 1  wherein the means for measuring the levels of said at least one pro-inflammatory cytokine in said biological sample comprises ELISAs, radioimmunoassays, multiplexed ELISAs on microarray platforms, multiplexed ELISAs using coded microspheres coupled with a flow cytometer detection systems, bioassays, Western blots, chromatograph-based separation systems, RT-PCR, competitive reverse transcription PCR, Northern blots, gene arrays, direct measurement of m-RNA, and mixtures thereof.  
     
     
         20 . The method according to  claim 19  wherein the means for measuring the levels of said at least one pro-inflammatory cytokine in said biological sample comprises ELISAs, RIAs, multiplexed ELISAs using coded microspheres coupled with a flow cytometer detection systems, and mixtures thereof.  
     
     
         21 . The method according to  claim 20 , wherein the means for measuring the levels of said at least one pro-inflammatory cytokine in said biological sample comprises multiplexed ELISAs using coded microspheres coupled with a flow cytometer detection systems.  
     
     
         22 . A kit comprising a first measuring element or system for measuring at least one anti-inflammatory cytokine in a biological sample from a mammalian subject before treatment and at at least one time point after or during treatment, a second measuring element or system for measuring at least one pro-inflammatory cytokine in a biological sample from said mammalian subject before treatment, and at at least one time point after or during treatment, wherein the change in ratio of anti-inflammatory to pro-inflammatory cytokine after administration of the treatment can be determined.  
     
     
         23 . The kit according to  claim 22 , wherein said kit further comprises language indicating that an increase in said ratio of anti-inflammatory cytokine to said pro-inflammatory cytokine is indicative of an inhibitor of inflammatory diseases of the bowel.