Immunosuppressive and tumour-suppressive bone marrow factor

Abstract

A novel immunosuppressive factor derived from mammalian bone marrow factor is described, which inhibits T lymphocyte activation and TNF-α production by activated macrophages and also inhibits tumor and leukemia cell growth. The factor provides a novel therapeutic composition for treatment of tumors and of disorders associated with inflammatory reactions or T lymphocyte activation.

1 . A purified, water soluble immunosuppressive factor characterised by a molecular weight less than 1000 daltons and movement as a single peak on reverse phase high performance liquid chromatography.  
     
     
         2 . The factor of    claim 1    having the following biological activities: 
 (a) prevention or reduction of IL-2 production by activated T lymphocytes;  
 (b) prevention or reduction of TNF-α production by activated macrophages;  
 (c) prevention or reduction of proliferation of activated T lymphocytes; and  
 (d) inhibition of mixed lymphocyte reaction.  
 
     
     
         3 . The factor of    claim 2    having the following further biological activity: 
 (a) inhibition of tumour growth in a mammal.  
 
     
     
         4 . The factor of    claim 3    having the following further biological activity: 
 (a) inhibition of leukemia cell proliferation.  
 
     
     
         5 . The factor of    claim 4    having the following further biological activity: 
 (a) inhibition of a graft versus host reaction in a mammal.  
 
     
     
         6 . The factor of    claim 2    wherein the factor is derived from mammalian bone marrow cells.  
     
     
         7 . The factor of    claim 6    wherein the bone marrow cells are myeloid progenitor cells.  
     
     
         8 . The factor of    claim 1    having a molecular weight of less than 1000 daltons.  
     
     
         9 . The factor of    claim 8    having a molecular weight of about 625 daltons.  
     
     
         10 . A method for inhibiting T lymphocyte activation in a mammal comprising administering to the mammal an effective amount of the factor of    claim 1   .  
     
     
         11 . A pharmaceutical composition for inhibiting T lymphocyte activation in a mammal comprising an effective amount of the factor of    claim 1    and a pharmaceutically acceptable carrier  
     
     
         12 . Use of a composition comprising an effective amount of the factor of    claim 1    and a pharmaceutically acceptable carrier to inhibit T lymphocyte activation in a mammal  
     
     
         13 . A method for treating or preventing a disorder associated with undesirable or excessive T lymphocyte activation, in a mammal, comprising administering to the mammal an effective amount of the factor of    claim 1     
     
     
         14 . The method of    claim 13    wherein the disorder is an autoimmune disease.  
     
     
         15 . The method of    claim 1    wherein the autoimmune disease is rheumatoid arthritis, lupus erythematosus, multiple sclerosis or type I diabetes  
     
     
         16 . The method of    claim 13    wherein the disorder involves allogeneic T lymphocyte interactions.  
     
     
         17 . The method of    claim 16    wherein the disorder is graft versus host disease.  
     
     
         18 . A pharmaceutical composition for treating or preventing a disorder associated with undesirable or excessive T lymphocyte activation comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier.  
     
     
         19 . Use of a composition comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier to treat or prevent a disorder associated with undesirable or excessive T lymphocyte activation.  
     
     
         20 . A method for inhibiting TNF-α production in a mammal comprising administering to the mammal an effective amount of the factor of any of    claims 1    to    9   .  
     
     
         21 . A pharmaceutical composition for inhibiting TNF-α production in a mammal coming an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier.  
     
     
         22 . Use of a composition comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier to inhibit TNF-α production in a mammal.  
     
     
         23 . A method for treating or preventing a disorder associated with undesirable or excessive TNF-α production, in a mammal, comprising administering to the mammal an effective amount of the factor of any of    claims 1    to    9   .  
     
     
         24 . A method for inhibiting growth of a tumour in a mammal comprising administering to the mammal an effective amount of the factor of any of    claims 1    to    9   .  
     
     
         25 . A pharmaceutical composition for inhibiting growth of a tumour in a mammal comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier.  
     
     
         26 . Use of a composition comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier to inhibit growth of a tumour in a mammal.  
     
     
         27 . A method for inhibiting growth of a leukemia in a mammal comprising administering to the mammal an effective amount of the factor of any of    claims 1    to    9   .  
     
     
         28 . A pharmaceutical composition for inhibiting growth of a leukemia in a mammal comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier.  
     
     
         29 . Use of a composition comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier to inhibit growth of a leukemia in a mammal.  
     
     
         30 . A method for preventing or treating, in a mammal, a disorder characterised by an abnormality in a signalling pathway, wherein the signalling pathway involves CD45 phosphatase activity, the method comprising administering to the mammal a therapeutically effective amount of the factor of any of    claims 1    to    9   .  
     
     
         31 . A method for preparing a purified immunosuppressive factor comprising the steps of 
 (a) culturing mammalian bone marrow cells to produce a factor-containing supernatant;    (b) subjecting the supernatant to solid phase extraction to produce a product;    (c) subjecting the product from step (b) to gel filtration to produce a product;    (d) subjecting the product from step (c) to anion exchange fast phase liquid chromatography to produce a product;    (e) subjecting the product from step (d) to amino-high performance liquid chromatography to produce a product; and    (f) subjecting the product from step (e) to reverse phase high performance liquid chromatography to product purified immunosuppressive factor.    
     
     
         32 . A purified immunosuppressive factor prepared in accordance with the method of    claim 31   .  
     
     
         33 . A method for inhibiting IL-2 production by activated T lymphocytes in a mammal comprising administering to the mammal an effective amount of the factor of any of    claims 1    to    9   .  
     
     
         34 . A pharmaceutical composition for inhibiting IL-2 production by activated T lymphocytes in a mammal comprising an effective amount of the factor of any of    claims 1    to    9    and a pharmaceutically acceptable carrier.