Cytolysis of target cells by superantigen conjugates inducing T-cell activation

Abstract

A method for inactivating target cells in the presence of T cells by bringing the two types of cells in contact with a superantigen (SAG) in the presence of an immune modulator, characterized in that at least one of the superantigen and the immune modulator is in the form of a conjugate between a “free” superantigen (Sag) and a moiety targeting the conjugate to the target cells. A superantigen conjugate complying with the formula (1): (T) x (Sag) y (IM) z ; a) T is a targeting moiety, Sag corresponds to a free superantigen, IM is an immune modulator that is not a superantigen and T, Sag and IM are linked together via organic linkers B; b) x, y and z are integers that typically are selected among 0-10 and represent the number of moieties T, Sag and IM, respetively, in a given conjugate molecule, with the provision that y>0 and also one or both of x and z>0. The superantigen conjugate is preferably a triple fusion protein. A targeted immune modulator, characterized in that it is a conjugate between a targeting moiety (T′″) and a modified immune modulator (IM′″). The conjugate complies with a formula analogous to formula (I) except for the imperative presence of the modified immune modulator. A superantigen moiety may be present. A DNA molecule encoding a superantigen and an immune modulator.

1 - 34 . (canceled)  
     
     
         35 . A pharmaceutical composition comprising a superantigen, an immune modulator and a targeting moiety, wherein at least one of the superantigen and immune modulator is conjugated to a targeting moiety.  
     
     
         36 . The pharmaceutical composition of  claim 35 , wherein the superantigen is conjugated to the targeting moiety.  
     
     
         37 . The pharmaceutical composition of  claim 35 , wherein the immune modulator is conjugated to the targeting moiety.  
     
     
         38 . The pharmaceutical composition of  claim 35 , wherein the superantigen and immune modulator are both conjugated to the same targeting moiety.  
     
     
         39 . The pharmaceutical composition of  claim 36 , wherein the immune modulator is not conjugated to the targeting moiety.  
     
     
         40 . The pharmaceutical composition of  claim 37 , wherein the superantigen is not conjugated to the targeting moiety.  
     
     
         41 . The pharmaceutical composition of  claim 35 , wherein the targeting moiety binds to a cell surface antigen.  
     
     
         42 . The pharmaceutical composition of  claim 36 , wherein the targeting moiety binds to a cell surface antigen.  
     
     
         43 . The pharmaceutical composition of  claim 41 , wherein the cell surface antigen is associated with a disease.  
     
     
         44 . The pharmaceutical composition of  claim 43 , wherein the disease is a cancer.  
     
     
         45 . The pharmaceutical composition of  claim 42 , wherein the cell surface antigen is associated with a disease.  
     
     
         46 . The pharmaceutical composition of  claim 45 , wherein the disease is a cancer.  
     
     
         47 . The pharmaceutical composition of  claim 35 , wherein the targeting moiety is selected from the group consisting of an antibody, an antigen-binding fragment of an antibody, an Fab fragment of an antibody, an Fab 2  fragment of an antibody, or a single chain antibody.  
     
     
         48 . The pharmaceutical composition of  claim 36 , wherein the targeting moiety is selected from the group consisting of an antibody, an antigen-binding fragment of an antibody, an Fab fragment of an antibody, an Fab 2  fragment of an antibody, or a single chain antibody.  
     
     
         49 . The pharmaceutical composition of  claim 46 , wherein the targeting moiety is selected from the group consisting of an antibody, an antigen-binding fragment of an antibody, an Fab fragment of an antibody, an Fab 2  fragment of an antibody, or a single chain antibody.  
     
     
         50 . The pharmaceutical composition of  claim 35 , wherein the superantigen is modified to have a decreased ability to bind to MHC class II antigen compared to a corresponding wild-type superantigen.  
     
     
         51 . The pharmaceutical composition of  claim 35 , wherein the superantigen is modified to have decreased seroreactivity in human sera compared to a corresponding wild-type superantigen.  
     
     
         52 . The pharmaceutical composition of  claim 35 , wherein the superantigen is chimeric comprising sequences derived from two or more wild type superantigens.  
     
     
         53 . The pharmaceutical composition of  claim 49 , wherein the superantigen is modified to have a decreased ability to bind to MHC class II antigen compared to a corresponding wild-type superantigen.  
     
     
         54 . The pharmaceutical composition of  claim 49 , wherein the superantigen is modified to have decreased seroreactivity in human sera compared to a corresponding wild-type superantigen.  
     
     
         55 . The pharmaceutical composition of  claim 49 , wherein the superantigen is chimeric comprising sequences derived from two or more wild type superantigens.  
     
     
         56 . The pharmaceutical composition of  claim 35 , wherein the superantigen is obtained from Staphylococcal enterotoxin.  
     
     
         57 . The pharmaceutical composition of  claim 56 , wherein the superantigen is obtained from Staphylococcal enterotoxin A.  
     
     
         58 . The pharmaceutical composition of  claim 57 , wherein the superantigen is Staphylococcal enterotoxin A.  
     
     
         59 . The pharmaceutical composition of  claim 49 , wherein the superantigen is obtained from Staphylococcal enterotoxin.  
     
     
         60 . The pharmaceutical composition of  claim 59 , wherein the superantigen is obtained from Staphylococcal enterotoxin A.  
     
     
         61 . The pharmaceutical composition of  claim 60 , wherein the superantigen is Staphylococcal enterotoxin A.  
     
     
         62 . The pharmaceutical composition of  claim 35 , wherein the immune modulator is selected from the group consisting of cytokines, chemokines, and extracellular parts of lymphocyte bound receptors and ligands.  
     
     
         63 . The pharmaceutical composition of  claim 35 , in the immune modulator is IL-2.  
     
     
         64 . The pharmaceutical composition of  claim 49 , wherein the immune modulator is selected from the group consisting of cytokines, chemokines, and extracellular parts of lymphocyte bound receptors and ligands.  
     
     
         65 . The pharmaceutical composition of  claim 49 , in the immune modulator is IL-2.  
     
     
         66 . A conjugate composition comprising a superantigen, an immune modulator and a targeting moiety, wherein at least one of the superantigen and immune modulator is conjugated to a targeting moiety.  
     
     
         67 . The conjugate composition of  claim 66 , wherein the superantigen is conjugated to the targeting moiety.  
     
     
         68 . The conjugate composition of  claim 66 , wherein the immune modulator is conjugated to the targeting moiety.  
     
     
         69 . The conjugate composition of  claim 66 , wherein the superantigen and immune modulator are both conjugated to the same targeting moiety.  
     
     
         70 . The conjugate composition of  claim 66 , wherein the targeting moiety binds to a cell surface antigen.  
     
     
         71 . The conjugate composition of  claim 70 , wherein the cell surface antigen is associated with a disease.  
     
     
         72 . The conjugate composition of  claim 71 , wherein the disease is a cancer.  
     
     
         73 . The conjugate composition of  claim 66 , wherein the targeting moiety is selected from the group consisting of an antibody, an antigen-binding fragment of an antibody, an Fab fragment of an antibody, an Fab 2  fragment of an antibody, or a single chain antibody.  
     
     
         74 . The conjugate composition of  claim 66 , wherein the superantigen is modified to have a decreased ability to bind to MHC class II antigen compared to a corresponding wild-type superantigen.  
     
     
         75 . The conjugate composition of  claim 66 , wherein the superantigen is modified to have decreased seroreactivity in human sera compared to a corresponding wild-type superantigen.  
     
     
         76 . The conjugate composition of  claim 66 , wherein the superantigen is chimeric comprising sequences derived from two or more wild type superantigens.  
     
     
         77 . The conjugate composition of  claim 66 , wherein the superantigen is obtained from Staphylococcal enterotoxin.  
     
     
         78 . The conjugate composition of  claim 77 , wherein the superantigen is obtained from Staphylococcal enterotoxin A.  
     
     
         79 . The conjugate composition of  claim 78 , wherein the superantigen is Staphylococcal enterotoxin A.  
     
     
         80 . The conjugate composition of  claim 66 , wherein the immune modulator is selected from the group consisting of cytokines, chemokines, and extracellular parts of lymphocyte bound receptors and ligands.  
     
     
         81 . The conjugate composition of  claim 66 , in the immune modulator is IL-2.