Immunomodulating tumor necrosis factor receptor 25 (TNFR25) agonists, antagonists and immunotoxins

Abstract

It is an object of the invention to provide novel compositions and methods utilizing immunomodulating agents that can either stimulate or indirectly augment the immune system or in other cases have an immunosuppressive effect. TNFR25 agonists disclosed herein have an anti-inflammatory and healing effect. They can be used, among other things, to treat disease caused by asthma and chronic inflammation such as for example inflammatory bowel diseases including ulcerative colitis and Crohn's Disease. TNFR25 antagonists disclosed herein are capable of inhibiting CD8 T cell-mediated cellular immune responses and can for example, mitigate organ or tissue rejection following a tissue transplantation. TNFR25 agonists disclosed herein represent biological response modifiers that alter the interaction between the body's cellular immune defenses and cancer cells to boost, direct, or restore the body's ability to fight the cancer when given with tumor vaccines. TNFR25 specific immunotoxins disclosed herein are also capable of increasing the effectiveness of a chemotherapeutic regimen by depleting a cancer patient of naturally occurring immunosuppressive cells.

1 . An isolated antibody that binds a Tumor Necrosis Factor Receptor 25 (TNFR25) antigen wherein said antibody is capable of acting as a TNFR25 agonist.  
     
     
         2 . The antibody of  claim 1 , wherein said antibody is capable of increasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody.  
     
     
         3 . The antibody of  claim 1 , wherein the Tumor Necrosis Factor Receptor 25 (TNFR25) antigen is human or mouse.  
     
     
         4 . A Tumor Necrosis Factor Receptor 25 (TNFR25)-specific toxin comprising a toxic agent linked to a polypeptide that binds the TNRF25 receptor.  
     
     
         5 . The toxin of  claim 4  wherein said polypeptide comprises monoclonal antibody 4C12 or an immunospecific portion thereof.  
     
     
         6 . The toxin of  claim 4  wherein said toxic agent is selected from the group consisting of a radioactive isotope, ricin, abrin, diphtheria toxin,  Pseudomonas  exotoxin, and metal ion.  
     
     
         7 . The toxin of  claim 4  wherein said polypeptide comprises a TL1A protein of a TNFR25-binding portion thereof.  
     
     
         8 . A method of activating Tumor Necrosis Factor Receptor 25 (TNFR25) receptor expressed on a cell comprising: 
 a. contacting the cell with an TNFR25 agonist, wherein said agonist is selected from the group consisting of: 
 i. a monoclonal antibody 4C12;  
 ii. an antibody that binds to a Tumor Necrosis Factor Receptor 25 (TNFR25), wherein said antibody is capable of increasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody;  
 iii. a soluble TL1A;  
 iv. an expression vector with an expression cassette capable of driving the transgenic expression of a TNFR25 agonist antibody;  
 v. an expression vector with an expression cassette capable of driving the transgenic expression of a soluble TL1A; and  
 vi. an expression vector with an expression cassette capable of driving the transgenic expression of a TNFR25;  
   b. observing an increase in TNFR25 receptor signaling.    
     
     
         9 . An isolated antibody that binds a Tumor Necrosis Factor Receptor 25 (TNFR25) antigen wherein said antibody is capable of acting as a TNFR25 antagonist.  
     
     
         10 . The antibody of  claim 9 , wherein said antibody binds a TL1A antigen, wherein said antibody is capable of decreasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody.  
     
     
         11 . The antibody of  claim 9 , wherein said Tumor Necrosis Factor Receptor 25 (TNFR25) antigen is human or mouse.  
     
     
         12 . A method of inhibiting Tumor Necrosis Factor Receptor 25 (TNFR25) receptor signaling in a cell comprising: 
 a. contacting the cell with a TNFR25 antagonist, wherein said antagonist is selected from the group consisting of: 
 i. a monoclonal antibody L4G6  
 ii. an antibody that binds to a Tumor Necrosis Factor Receptor 25 (TNFR25) wherein said antibody is capable of decreasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody;  
 iii. an antibody specific for soluble TL1A;  
 iv. a fusion protein comprising an extracellular portion of TNFR25;  
 v. an expression vector with an expression cassette capable of driving the transgenic expression of the extracellular portion of TNFR25;  
 v. an expression vector with an expression cassette capable of driving the transgenic expression of an antibody specific for soluble TL1A; and  
 vi. an expression vector with an expression cassette capable of driving the transgenic expression of L4G6;  
   b. observing an decrease in TNFR25 receptor signaling.    
     
     
         13 . A tumor vaccine comprising a tumor antigen and a TNFR25 agonist as a biological response modifier.  
     
     
         14 . The tumor vaccine of  claim 12  wherein said TNFR25 agonist is selected from the group consisting of consisting of: 
 a. a monoclonal antibody 4C12;    b. an antibody that binds to a Tumor Necrosis Factor Receptor 25 (TNFR25), wherein said antibody is capable of increasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody;    c. a soluble TL1A;    d. an expression vector with an expression cassette capable of driving the transgenic expression of a TNFR25 agonist antibody;    e. an expression vector with an expression cassette capable of driving the transgenic expression of a soluble TL1A; and    f. an expression vector with an expression cassette capable of driving the transgenic expression of a TNFR25.    
     
     
         15 . The tumor vaccine of  claim 14  further comprising an adjuvant.  
     
     
         16 . A method of immunizing a patient against a tumor comprising isolating a tumor specific antigen from said tumor; and administering to the patient a tumor vaccine of  claim 14 .  
     
     
         17 . A method of treating cancer in a patient comprising depleting a patient of CD4+/CD25+T regulatory cells (Tregs) by providing the patient with a composition comprising the toxin of  claim 4;  and providing a patient with a chemotherapeutic agent.  
     
     
         18 . A method of treating and/or preventing gut inflammation comprising providing a patient in need thereof a with an effective amount of a therapeutic composition comprising a TNFR25 agonist.  
     
     
         19 . The method of  claim 18 , wherein said gut inflammation condition is selected from the group consisting of inflammatory bowel disease, Crohn's disease, and ulcerative colitis.  
     
     
         20 . The method of  claim 18 , wherein said TNFR25 agonist is selected from the group consisting of 
 a. soluble TL1A    b. a monoclonal antibody 4C12    c. an expression vector with an expression cassette capable of driving the transgenic expression of a TNFR25 agonist antibody;    d. an expression vector with an expression cassette capable of driving the transgenic expression of a soluble TL1A; and    e. an expression vector with an expression cassette capable of driving the transgenic expression of a TNFR25.    
     
     
         21 . A therapeutic composition for the facilitation of an organ transplant comprising a TNFR25 antagonist and an immunosuppressant.  
     
     
         22 . The composition of  claim 21  wherein the immunosuppressant is selected from the group consisting of glucocorticoid, azathioprine, methotrexate, cyclosporin, cyclophosphamide, mercaptopurine, tacrolimus and mycophenolate mofetil.  
     
     
         23 . The composition of  claim 21  wherein said TNFR25 antagonist is selected from the group consisting of: 
 a. a monoclonal antibody L4G6    b. an antibody that binds to a Tumor Necrosis Factor Receptor 25 (TNFR25 is capable of decreasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody;    c. an antibody specific for soluble TL1A;    d. a fusion protein comprising an extracellular portion of TNFR25;    e. an expression vector with an expression cassette capable of driving the transgenic expression of the extracellular portion of TNFR25;    f. an expression vector with an expression cassette capable of driving the transgenic expression of an antibody specific for soluble TL1A; and    g. an expression vector with an expression cassette capable of driving the transgenic expression of L4G6.    
     
     
         24 . A method of transplanting a tissue from a donor into a host comprising: 
 a. obtaining the tissue from the donor;    b. providing the host with the composition of  claim 21;     c. transplanting the tissue into the host.    
     
     
         25 . A method of inhibiting the clonal expansion of a population of cognate CD8 T cells comprising, 
 a. exposing the CD8 T cells to their cognate antigen; and    b. exposing the CD8 T cells to a TNFR25 antagonist.    
     
     
         26 . The method of  claim 25  wherein said TNFR25 antagonist is selected from the group consisting of: 
 a. a monoclonal antibody L4G6    b. an antibody that binds to a Tumor Necrosis Factor Receptor 25 (TNFR25) wherein said antibody is capable of decreasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody;    c. an antibody specific for soluble TL1A;    d. a fusion protein comprising an extracellular portion of TNFR25;    e. an expression vector with an expression cassette capable of driving the transgenic expression of the extracellular portion of TNFR25;    f. an expression vector with an expression cassette capable of driving the transgenic expression of an antibody specific for soluble TL1A; and    g. an expression vector with an expression cassette capable of driving the transgenic expression of L4G6.    
     
     
         27 . The method of  claim 26  wherein cognate antigen is associated with tissue to be transplanted from a donor into a host.  
     
     
         28 . A method of treating and/or preventing lung inflammation comprising providing a patient in need thereof with an effective amount of a therapeutic composition comprising a TNFR25 antagonist.  
     
     
         29 . The method of  claim 28 , wherein said inflammation condition is a result of asthma.  
     
     
         30 . The method of  claim 28  wherein said TNFR25 antagonist is selected from the group consisting of: 
 a. a monoclonal antibody L4G6    b. an antibody that binds to a Tumor Necrosis Factor Receptor 25 (TNFR25) wherein said antibody is capable of decreasing OT-I CD8 cell expansion when cross-primed by gp96-Ig-ovalbumin relative to a control antibody;    c. an antibody specific for soluble TL1A;    d. a fusion protein comprising an extracellular portion of TNFR25;    e. an expression vector with an expression cassette capable of driving the transgenic expression of the extracellular portion of TNFR25;    f. an expression vector with an expression cassette capable of driving the transgenic expression of an antibody specific for soluble TL1A; and    g. an expression vector with an expression cassette capable of driving the transgenic expression of L4G6.    
     
     
         31 . An isolated TNFR25 antagonist comprising a polypeptide encoded by a nucleic acid comprising a sequence that hybridizes under stringent conditions to SEQ ID NOs: 4, 5, 6 and/or 16, and wherein said sequence encodes an amino acid sequence capable of binding a TL1A protein.  
     
     
         32 . The isolated TNFR25 antagonist of  claim 31 , wherein said sequence hybridizes under stringent conditions to SEQ ID NO: 4.  
     
     
         33 . The isolated TNFR25 antagonist of  claim 31 , wherein said sequence hybridizes under stringent conditions to SEQ ID NO: 5.  
     
     
         34 . The isolated TNFR25 antagonist of  claim 31 , wherein said sequence hybridizes under stringent conditions to SEQ ID NO: 6.  
     
     
         35 . The isolated TNFR25 antagonist of  claim 31 , wherein said TL1A is human TL1A.  
     
     
         36 . The isolated TNFR25 antagonist of  claim 31 , wherein said TL1A is mouse TL1A.  
     
     
         37 . A method of treating and/or preventing lung inflammation comprising providing a patient in need thereof with an effective amount of a therapeutic composition comprising the TNFR25 antagonist of  claim 31 .  
     
     
         38 . A method of transplanting a tissue from a donor into a host comprising: 
 a. obtaining the tissue from the donor;    b. providing the host with the TNFR25 antagonist of  claim 31;     c. transplanting the tissue into the host.    
     
     
         39 . A composition comprising a polypeptide encoded by a sequence that hybridizes under stringent conditions to SEQ ID NOs: 3 and/or 7, and wherein said sequence encodes an amino acid sequence capable of binding a TNFR25 receptor protein; and a toxic agent.  
     
     
         40 . The composition of  claim 39  wherein said toxic agent is selected from the group consisting of a radioactive isotope, ricin, abrin, diphtheria toxin,  Pseudomonas  exotoxin, and metal ion.  
     
     
         41 . A method of treating cancer in a patient comprising depleting a patient of CD4+/CD25+T regulatory cells (Tregs) by providing the patient with a composition comprising the toxin of  claim 39;  and providing a patient with a chemotherapeutic agent.  
     
     
         42 . A method of treating and/or preventing gut inflammation comprising providing a patient in need thereof a with an effective amount of a composition comprising a polypeptide encoded by a sequence that hybridizes under stringent conditions to SEQ ID NOs: 3 and/or 7, and wherein said sequence encodes an amino acid sequence capable of binding a TNFR25 receptor protein.  
     
     
         43 . The method of  claim 42  wherein said gut inflammation associated with irritable bowel syndrome.  
     
     
         44 . The method of  claim 42  wherein said gut inflammation associated with Crohn's disease.  
     
     
         45 . A tumor vaccine comprising a tumor antigen and a polypeptide encoded by a sequence that hybridizes under stringent conditions to SEQ ID NOs: 3 and/or 7, and wherein said sequence encodes an amino acid sequence capable of binding a TNFR25 receptor protein, as a biological response modifier.  
     
     
         46 . An expression vector comprising a nucleic acid sequence that hybridizes under stringent conditions to SEQ ID NOs: 3 and/or 7, and wherein said sequence encodes an amino acid sequence capable of binding a TNFR25 receptor protein.  
     
     
         47 . An expression vector comprising a nucleic acid sequence that hybridizes under stringent conditions to SEQ ID NOs: 4, 5, 6 and/or 16, and wherein said sequence encodes an amino acid sequence capable of binding a TL1A protein