Preparation of potent macrophage activating factors derived from cloned vitamin D binding protein and its domain and their therapeutic usage for cancer, HIV-infection and osteopetrosis

Abstract

Vitamin D-binding protein (Gc protein) and its small domain (approximately ⅕ of the Gc peptide also known as domain III) were cloned via a baculovirus vector. The cloned Gc protein and the cloned domain (Cd) peptide were treated with immobilized β-galactosidase and sialidase to yield macrophage activating factors, GcMAFc and CdMAF, respectively. These cloned macrophage activating factors and GcMAF are to be used for therapy of cancer, HIV-infection and osteopetrosis, and may also be used as adjuvants for immunization and vaccination.

I claim:  
     
         1 . A process for cloning vitamin D 3 -binding protein (Gc protein) into baculovirus comprising the step of selecting and using a baculovirus vector to clone the vitamin D 3 -binding protein Gc protein (Gc protein).  
     
     
         2 . A process for producing a cloned macrophage activating factor (GcMAFc) comprising contacting cloned Gc protein in vitro with immobilized β-galactosidase and sialidase and obtaining the cloned macrophage activating factor (GcMAFc).  
     
     
         3 . A process for cloning vitamin D 3 -binding protein domain III (Gc domain III) into baculovirus comprising the step of selecting and utilizing a baculovirus vector to clone the vitamin D 3 -binding protein domain III (Gc domain III).  
     
     
         4 . A process for producing a cloned macrophage activating factor (CdMAF) comprising contacting cloned Gc domain III in vitro with immobilized β-galactosidase and sialidase and obtaining the macrophage activating factor (CdMAF).  
     
     
         5 . A method of treating a person suffering from cancer by administering to the person a therapeutically effective amount of a Gc protein macrophage activating factor (GcMAF), the GcMAF being a product of contacting serum Gc protein in vitro with immobilized β-galactosidase and sialidase.  
     
     
         6 . A method of treating a person suffering from cancer by administering to the person a therapeutically effective amount of a cloned macrophage activating factor (GcMAFc), which is a product of the process according to  claim 2 .  
     
     
         7 . A method of treating a person suffering from cancer by administering to the person a therapeutically effective amount of a cloned macrophage activating factor (CdMAF), which is a product of the process according to  claim 4 .  
     
     
         8 . A method of treating a person suffering from human immunodeficiency virus (HIV), Epstein-Barr virus (EBV) or herpes zoster by administering to the person a therapeutically effective amount of a macrophage activating factor (GcMAF), which is a product of contacting serum Gc protein in vitro with immobilized β-galactosidase and sialidase.  
     
     
         9 . A method of treating a person suffering from human immunodeficiency virus (HIV), Epstein-Barr virus (EBV) or herpes zoster by administering to the person a therapeutically effective amount of a macrophage activating factor (GcMAFc), which is a product of the process according to  claim 2 .  
     
     
         10 . A method of treating a person suffering from human immunodeficiency virus (HIV), Epstein-Barr virus (EBV) or herpes zoster by administering to the person a therapeutically effective amount of a macrophage activating factor (CdMAF), which is a product of the process according to  claim 4 .  
     
     
         11 . A macrophage activating factor (GcMAFc), which is a product of the process according to  claim 2 .  
     
     
         12 . A macrophage activating factor (CdMAF), which is a product of the process according to  claim 4 .  
     
     
         13 . A method of promoting bone marrow formation in osteopetrotic patients comprising administering a therapeutically effective amount of a macrophage activating factor (GcMAFc), which is a product of the process according to  claim 2 .  
     
     
         14 . A method of promoting bone marrow formation in osteopetrotic patients comprising administering a therapeutically effective amount of a macrophage activating factor (CdMAF), which is a product of the process according to  claim 4 .  
     
     
         15 . An adjuvant for immunizing humans and animals with antigens or vaccines, the adjuvant comprising a macrophage activating factor (GcMAF) which is a product of contacting serum Gc protein in vitro with immobilized β-galactosidase and sialidase.  
     
     
         16 . An adjuvant for immunizing humans and animals with antigens or vaccines, the adjuvant comprising a macrophage activating factor (GcMAFc), which is a product of the process according to  claim 2 .  
     
     
         17 . An adjuvant for immunizing humans and animals with antigens or vaccines, the adjuvant comprising a macrophage activating factor (CdMAF), which is a product of the process according to  claim 4 .  
     
     
         18 . A cloned vitamin D 3 -binding protein (Gc protein) having an amino acid sequence of FIG. 3 (SEQ. ID. NO:1)(GcMAFc).  
     
     
         19 . A cloned vitamin D 3 -binding protein domain III (Gc domain III) having an amino acid sequence of FIG. 5 (SEQ ID. NO:2)(CdMAF 1 ).  
     
     
         20 . A cloned vitamin D 3 -binding protein domain III (Gc domain II) having an amino acid sequence of FIG. 7 (SEQ ID. NO:3)(CdMAF 2 ).