Compositions and methods for treatment of neoplastic disease

Abstract

The present invention comprises compositions and methods for treating a tumor or neoplastic disease in a host, The methods employ conjugates comprising superantigen polypeptides or nucleic acids with other structures that preferentially bind to tumor cells and are capable of inducing apoptosis. Also provided are superantigen-glycolipid conjugates and vesicles that are loaded onto antigen presenting cells to activate both T cells and NKT cells. Cell-based vaccines comprise tumor cells engineered to express a superantigen along with glycolipids products which, when expressed, render the cells capable of eliciting an effective anti-tumor immune response in a mammal into which these cells are introduced. Included among these compositions are tumor cells, hybrid cells of tumor cells and accessory cells, preferably dendritic cells. Also provided are T cells and NKT cells activated by the above compositions that can be administered for adoptive immunotherapy.

1 . A mammalian cell comprising an exogenous nucleic acid encoding a superantigen which is expressed in said cell which cell also produces or expresses all α-anomers of monoglycosylceramide or diglycosylceramide, wherein expression of said superantigen and said mono- or di-glycosylceramide is capable of eliciting an effective anti-tumor immune response in a mammal into which said cell is introduced.  
     
     
         2 . The cell of  claim 1  which is selected from a group consisting of 
 (a) a tumor cell    (b) an accessory cell    (c) a tumor cell/accessory cell hybrid    
     
     
         3 . The cell of  claim 1  wherein said mammal bears a tumor, against which the antitumor response is directed, said tumor being selected from the group consisting of a carcinoma, a melanoma, a sarcoma, a neuroblastoma, an astrocytoma, a lymphoma and a leukemia.  
     
     
         4 . The cell of  claim 1  wherein the superantigen is selected from the group consisting of a Staphylococcal enterotoxin and a Streptococcal pyrogenic exotoxin.  
     
     
         5 . A method of treating a tumor or neoplastic disease in a subject, comprising administering to said subject an effective amount of the cells of any of claims  1 - 4 , wherein said nucleic acid is introduced in vivo into a cell that produces or expresses said mono- or diglycosylceramide.  
     
     
         6 . A method of treating a tumor or neoplastic disease in a subject comprising administering an effective amount of the cells of any of claims  1 - 4  wherein said superantigen exogenous nucleic acids is introduced ex vivo or in vitro into a cell which produces or expresses said mono- or di-glycosylceramide.  
     
     
         7 . A composition useful for treating a tumor or neoplastic disease in a subject comprising a conjugate or complex of 
 (a) a superantigen; and    (b) a glycosylceramide.    
     
     
         8 . The composition of  claim 7  wherein the glycosylceramide is selected from a group consisting of 
 (a) an α-1-4-galabiosylceramide,    (b) an α-1-4-globotriosylceramide,    (c) an α-1-4-globoteterasylceramide or    (d) a glycosylphosphatidylinositol-anchored α-1-4-galabiosylceramide,    (e) a glycosylphosphatidylinositol-anchored α-1-4-globotriosylceramide, and    (f) a glycosylphosphatidylinositol-anchored α-1-4-globoteterasylceramide.    
     
     
         9 . The composition of  claim 7 , wherein the glycosylceramide comprises a phytosphingosine chain having unsubstituted hydroxyl groups at its C3- and C4 position.  
     
     
         10 . The composition of  claim 7 , wherein the length of the ceramide fatty acyl chain is from about 12 to about 24 carbons.  
     
     
         11 . The composition of  claim 7 , wherein the sphingosine portion of the ceramide has a chain length of about 10 to about 13 carbons.  
     
     
         12 . The composition of  claim 7 , wherein the conjugate or complex further comprises CD1 receptors, MHC class I molecules, MHC class II molecules or superantigen receptors.  
     
     
         13 . The composition of claim  7 - 12  wherein the superantigen-glycosylceramide is in or on a vesicle, exosome, liposome, phage display, prokaryotic cell surface or eukaryotic cell surface.  
     
     
         14 . The composition of claims  7 - 12  wherein said complex or conjugate is obtained by shedding from cells expressing said complexes or conjugates.  
     
     
         15 . The composition of claim  7 - 12  wherein the superantigen-glycosylceramide conjugate, or, if present, said superantigen-GPI-glycosylceramide conjugate, is chemically linked by a crosslinking agent  
     
     
         16 . The compositions of claims  13  wherein the superantigen-glycosylceramide is loaded onto a prokaryotic or eukaryotic cell surface.  
     
     
         17 . A method of treating a tumor or neoplastic disease in a subject comprising administering to the subject an effective amount of cells loaded with the compositions of any of claims  7 - 12 , so that the cells present the composition to the immune system, thereby inducing an anti-tumor immune response.  
     
     
         18 . A method of treating a tumor or neoplastic disease in a subject comprising administering to the subject an effective amount of compositions of  claim 13 , thereby inducing an anti-tumor immune response.  
     
     
         19 . A method of treating a tumor or neoplastic disease in a subject comprising administering to the subject an effective amount of the composition of  claim 15 , thereby inducing an anti-tumor immune response.  
     
     
         20 . The method of  claim 17  wherein the composition is a superantigen-glycosylceramide-CD1 conjugate or complex wherein the glycosylceramide is selected from a group consisting of 
 (a) an α1-4-galabiosylceramide,    (b) an α-1-4-globotriosylceramide,    (c) an α-1-4-globoteterasylceramide or    (d) a glycosylphosphatidylinositol-anchored α-1-4-galabiosylceramide,    (e) a glycosylphosphatidylinositol-anchored α-1-4-globotriosylceramide, and    (f) a glycosylphosphatidylinositol-anchored α-1-4-globoteterasylceramide.    
     
     
         21 . The method of  claim 18  wherein the composition is a superantigen-glycosylceramide-CD1 conjugate or complex wherein the glycosylceramide is selected from a group consisting of 
 (a) an α1-4-galabiosylceramide,    (b) an α-1-4-globotriosylceramide,    (c) an α-1-4-globoteterasylceramide or    (d) a glycosylphosphatidylinositol-anchored α-1-4-galabiosylceramide,    (e) a glycosylphosphatidylinositol-anchored α-1-4-globotriosylceramide, and    (f) a glycosylphosphatidylinositol-anchored α-1-4-globoteterasylceramide.    
     
     
         22 . The method of  claim 17  wherein the cells are selected from the group consisting of: 
 (a) dendritic cells;    (b) macrophages or monocytes;    (c) fibroblasts;    (d) keratinocytes;    (e) stromal cells;    (f) antigen presenting cell;    (g) tumor cells;    (h) lymphocytes; and    (i) a combination of any two or more of (a)-(h).    
     
     
         23 . The method of  claim 18  wherein the cells are selected from the group consisting of: 
 (a) dendritic cells;    (b) macrophages or monocytes;    (c) fibroblasts;    (d) keratinocytes;    (e) stromal cells;    (f) antigen presenting cell;    (g) tumor cells;    (h) lymphocytes; and    (i) a combination of any two or more of (a)-(h).    
     
     
         24 . The method of  claim 19  wherein the cells are selected from the group consisting of: 
 (a) dendritic cells;    (b) macrophages or monocytes;    (c) fibroblasts;    (d) keratinocytes;    (e) stromal cells;    (f) antigen presenting cell;    (g) tumor cells;    (h) lymphocytes; and    (i) a combination of any two or more of (a)-(h).    
     
     
         25 . The method of  claim 20  wherein the cells are selected from the group consisting of: 
 (a) dendritic cells;    (b) macrophages or monocytes;    (c) fibroblasts;    (d) keratinocytes;    (e) stromal cells;    (f) antigen presenting cell;    (g) tumor cells;    (h) lymphocytes; and    (i) a combination of any two or more of (a)-(h).    
     
     
         26 . The compositions of claim  7 - 12 , wherein the superantigen is selected from the group consisting of a Staphylococcal enterotoxin and a Streptococcal pyrogenic exotoxin.  
     
     
         27 . A method of treating a tumor or neoplastic disease in a subject comprising administering to said subject an effective amount of the composition of any of claims.  
     
     
         28 . A method of preparing a population of immunotherapeutically active T or NKT cells useful to treat a tumor or neoplastic disease in a subject, comprising: 
 (a) providing to 
 (i) a subject in vivo or  
 (ii) a population of T and/or NKT cells ex vivo or in vitro  
    the cells of any of claims  1 - 4  or a composition of any of claims  7 - 12  to prime or stimulate the production of a population of tumor-specific T cells and/or NKT cells,    (b) obtaining said primed or stimulated T or NKT cells;    (c) optionally, further contacting said primed or stimulated T or NKT cells with any of said cells or compositions ex vivo to expand and further stimulate said T or NKT cells.    thereby preparing said of immunotherapeutically active cells.    
     
     
         29 . A method of treating a tumor or neoplastic disease in a subject, comprising administering an effective amount of T and/or NKT cells prepared in accordance with  claim 24  to said subject to treat said tumor or neoplastic disease.  
     
     
         30 . A composition useful for treating a tumor or neoplastic disease in a subject comprising naked DNA encoding a superantigen conjugated to a protein which induces apoptosis of tumor cells in said subject.  
     
     
         31 . The composition of  claim 25  wherein the protein is selected from a group consisting of 
 (a) Fas;    (b) Perforin;    (c) Granzyme B;    (d) Tumor Necrosis Factor α or □;    (e) Verotoxin; and    (f) a Verotoxin A chain, B chain or hybrid AB chain.    
     
     
         32 . A composition useful for treating a tumor or neoplastic disease in a subject comprising naked DNA encoding a superantigen conjugated to a verotoxin.  
     
     
         33 . A composition useful for treating a tumor or neoplastic disease in a subject comprising naked DNA encoding a superantigen conjugated to a protein or peptide that has at least about 30% sequence identity to the Gal(α1-4)Gal-binding region of a verotoxin.  
     
     
         34 . A composition useful for treating a tumor or neoplastic disease in a subject comprising naked DNA encoding a superantigen conjugated to a protein or peptide that has at least about 45% sequence identity to the Gal(α1-4)Gal-binding region of a verotoxin.  
     
     
         35 . The composition of  claim 33  or  34  wherein the peptide or protein has the amino acid sequence of all or part of any Gal(α1-4)Gal-binding portion of: 
 (a) the 63 kDa extracellular peptide of the interferon α receptor; or    (b) the N terminal extracellular domain of CD19.    
     
     
         36 . An apoptotic cell preparation or lysate useful for treating a tumor or neoplastic disease in a subject, comprising a cell population that has been 
 (a) transfected with naked DNA encoding a superantigen; and    (b) treated to undergo apoptosis or lysis.    
     
     
         37 . A cell which has ingested or been transfected with the apoptotic preparation or lysate of  claim 36 , thereby rendering the cell effective in presenting material expressed from transfecting nucleic acid or material ingested to the immune system of a mammal to elicit an anti-tumor immune response.  
     
     
         38 . A method for treating a tumor or neoplastic disease in a subject, comprising 
 (a) providing to a population of cells selected from the group consisting of: 
 (i) tumor cells;  
 (ii) accessory cells;  
 (iii) tumor cell/accessory cell hybrids;  
 (iv) cells with an inherent or acquired -galactosidase deficiency; and  
 (v) a combination of any two or more of (i)-(iv),  
 said apoptotic cell preparation or said lysate of  claim 36 , to produce an immunostimulatory cell population;  
   (b) administering to said subjected an amount of said immunostimulatory cell population effective to treat said tumor or neoplastic disease.    
     
     
         39 . The method of  claim 38  wherein said accessory cells are dendritic cells and said hybrid cells (iii) are dendritic cell/tumor cell hybrids.  
     
     
         40 . The method of  claim 38  wherein the providing step (a) is in vivo.  
     
     
         41 . The method of  claim 38  wherein the providing step (a) is in vitro.  
     
     
         42 . A method of treating a tumor or neoplastic disease in a subject comprising administering to said subject an effective amount of cells according to  claim 36 , wherein said ingested lysate, transfecting nucleic acid or other apoptotic matter is presented to the immune system to elicit a tumoricidal response.  
     
     
         43 . A composition useful for treating a tumor or neoplastic disease in a subject comprising a lipoprotein which is capable of binding to receptors in tumor microvasculature and eliciting apoptosis of tumor endothelial cells and eliciting an effective anti-tumor response in a mammal into which said lipoprotein is introduced.  
     
     
         44 . The composition of  claim 43  wherein the lipoprotein is selected from the group consisting of: 
 (a) low density lipoproteins    (a) chylomicrons    (b) very low density lipoproteins    (c) apolipoproteins    (d) oxidized low density lipoproteins    (e) oxidized low density lipoprotein byproducts    (f) oxidized low density lipoproteins mimics    (g) low density lipoprotein complexed with compounds which enhance or promote the uptake by cells expressing LDL or oxidized LDL receptors.    
     
     
         45 . The composition of  claim 44  wherein the compounds which enhance or promote the uptake by cells expressing LDL or oxidized LDL receptors are selected from a group consisting of: 
 (a) fibronectin    (b) collagen    (c) heparan    
     
     
         46 . A composition useful for treating a tumor or neoplastic disease in a subject comprising a conjugate or complex of: 
 (a) a superantigen; and    (b) a lipoprotein    
     
     
         47 . The composition of  claim 46  wherein the lipoprotein is selected from the group consisting of: 
 (a) low density lipoproteins    (a) chylomicrons    (b) very low density lipoproteins    (c) apolipoproteins    (d) oxidized low density lipoproteins    (e) oxidized low density lipoprotein byproducts    (f) oxidized low density lipoproteins mimics    (g) low density lipoprotein complexed with compounds which enhance or promote the uptake by cells expressing LDL or oxidized LDL receptors.    
     
     
         48 . the composition of  claim 47  wherein the compounds which enhance or promote the uptake by cells expressing LDL or oxidized LDL receptors are selected from a group consisting of: 
 (a) fibronectin    (b) collagen    (c) heparan    
     
     
         49 . The compositions of claims  46 - 48  wherein the superantigen-lipoprotein conjugate is in or on a vesicle, exosome, liposome, phage display, prokaryotic cell surface or eukaryotic cell surface.  
     
     
         50 . The compositions of  claim 49  which are are derived from a group consisting of: 
 (a) a mammalian cell transfected with superantigen genes    (b) a sickle cell or sickle cell precursor transfected with superantigen genes    (c) a yeast cell or mutant transfected with superantigen genes    (d) a  Staphylococcus carnosus  transfected with superantigen genes.    (e) a  Sphingomonas paucimobilis  transfected with superantigen genes    
     
     
         51  The compositions of claims  46 - 50  wherein the superantigen is selected from the group consisting of a Staphylococcal enterotoxin and a Streptococcal pyrogenic exotoxin.  
     
     
         52 . The method of treating a tumor or neoplastic disease in a subject comprising administering to the subject an effective amount of the compositions of claims  46 - 50  so that the composition localizes in tumor microvasuclature and is presented to the immune system, thereby inducing an anti-tumor response  
     
     
         53 . A mammalian cell useful for treating a tumor or neoplastic disease in a subject comprising a nucleic acid encoding a receptor for LDL or oxidized LDL which renders the said cell capable of binding LDL or oxyLDL and undergoing apoptosis and eliciting an effective anti-tumor response.  
     
     
         54 . The mammalian cell of  claim 53  comprising a second exogenous nucleic acid encoding a superantigen such that the expression of said superantigen and products of the first nucleic acid alone or in combination are capable of eliciting an effective anti-tumor immune response.  
     
     
         55 . The cell of  claim 53  is selected from a group consisting of: 
 (a) tumor cells    (b) endothelial cells    (c) stromal cells    
     
     
         56 . The cell of  claim 53  wherein the LDL or oxidized LDL receptor is selected for the group consisting of: 
 (a) scavenger receptors expresssed on endothelial cells and macrophages    (b) LOX-1 receptor    (c) oxidized low density lipoprotein receptor    (d) CD36 receptor    (e) Acetyl low density lipoprotein receptor    (f) low density lipoprotein receptor    (g) low density lipoprotein receptor-related protein (LRP)    
     
     
         57 . The cell of  claim 55  wherein the superantigen is selected from the group consisting of a Staphylococcal enterotoxin and a Streptococcal pyrogenic exotoxin.  
     
     
         58 . A method of treating a tumor or neoplastic disease in a subject comprising administering an effective amount of exogenous LDL receptor, oxyLDL receptor nucleic acid or superantigen nucleic acid wherein they are introduced into the cell in vivo.  
     
     
         59 . A composition useful for treating a tumor or neoplastic disease in a subject comprising a conjugate or complex of: 
 (a) a superantigen; and    (b) a LDL or oxidized low density lipoprotein receptor    
     
     
         60 . The composition of  claim 59  wherein the LDL or oxidized LDL receptor is selected for the group consisting of: 
 (a) scavenger receptors expresssed on endothelial cells and macrophages    (b) LOX-1 receptor    (c) oxidized low density lipoprotein receptor    (d) CD36 receptor    (e) Acetyl low density lipoprotein receptor    (f) low density lipoprotein receptor    (g) low density lipoprotein receptor-related protein (LRP)    (h) apolipoprotein receptors    
     
     
         61 . The compositions of claims  58 - 59  wherein the LDL and oxidized LDL receptors are in the form of naked DNA.  
     
     
         62 . The compositions of claims  59 - 60  wherein the LDL receptor or superantigen or LDL receptor is a polypeptide or nucleic acid in or on a vesicle, exosome, liposome, phage display, plasmid, expression vector, prokaryotic cell surface or eukaryotic cell surface.  
     
     
         63 . The vesicles, exosomes prokaryotic and eukaryotic cell surfaces of  claim 61  which are derived from a group consisting of: 
 (a) a mammalian cell transfected with superantigen genes    (b) a sickle cell or sickle cell precursor transfected with superantigen genes    (c) a yeast cell or mutant yeast cell transfected with superantigen genes    (d) a  Staphylococcus carnosus  transfected with superantigen genes.    (e) a  Sphingomonas paucimobilis  transfected with superantigen genes    
     
     
         64 . The compositions of claims  58 - 63  wherein the superantigen is selected from the group comprising a Staphylococcal enterotoxin and Streptococcal pyrogenic exotoxin  
     
     
         65 . A mammalian cell comprising an exogenous nucleic acid encoding a superantigen which is expressed in said cell which cell also produces or expresses low density lipoproteins, wherein expression of said superantigen and said native LDL or oxidized LDL or biologically active LDL mimics and byproducts is capable of eliciting an effective anti-tumor immune response in a mammal into which said cell is introduced.  
     
     
         66 . The cell of  claim 65  which is selected from a group consisting of 
 (a) a tumor cell    (b) a endothelial cell    (b) a sickled cell or sickled cell precursor    
     
     
         67 . The cell of  claim 65  wherein the the low density lipoproteins are selected from the group consisting of: 
 (a) native low density lipoprotein    (a) oxidized low density lipoprotein    (b) low density lipoprotein mimics    (c) low density lipoprotein byproducts    
     
     
         68 . The cell of  claim 65  wherein said mammal bears a tumor, against which the antitumor response is directed, said tumor being selected from the group consisting of a carcinoma, a melanoma, a sarcoma, a neuroblastoma, an astrocytoma, a lymphoma and a leukemia.  
     
     
         69 . A method of treating a tumor or neoplastic disease in a subject, comprising administering to said subject an effective amount of the cells of any of claims.  
     
     
         70 . A method of treating a tumor or neoplastic disease in a subject comprising administering an effective amount of the cells of any of claims  65 - 67  wherein said superantigen exogenous nucleic acids is introduced ex vivo or in vitro into a cell which produces or expresses low density lipoproteins  
     
     
         71 . A method of treating a tumor or neoplastic disease in a subject comprising administering an effective amount of the cells of any of claims  65 - 67  wherein said superantigen exogenous nucleic acids and apolipoprotein nucleic acid is introduced ex vivo or in vitro into a cell which thereby expresses superantigen and apolipoprotein.  
     
     
         72 . The method of claim  70 - 71  wherin the low density lipoproteins are selected from a group consisting of: 
 (a) native low density lipoprotein    (a) oxidized low density lipoprotein    (b) low density lipoprotein mimics    (c) low density lipoprotein byproducts    
     
     
         73 . A mammalian cell comprising expressing a α-monogalactosylceramide or α-digalactosylceramide or oxidized low density lipoprotein individually or in any combination which is (are) capable of binding to tumor microvasculature, inducing apoptosis and eliciting an effective anti-tumor immune response in a mammal into which said cell is introduced.  
     
     
         74 . The cell of  claim 73  which also expresses a superantigen  
     
     
         75 . The cell of claims  73 - 74  which is selected from a group consisting of: 
 (a) a tumor cell    (b) a sickled cell or sickled cell precursor    
     
     
         76 . A mammalian tumor cell/accessory cell hybrid cell comprising an exogenous nucleic acid encoding a superantigen which is expressed in said cell such that expression of said superantigen renders the said cell capable of elicting an effective anti-tumor immune response in a mammal into which said cell is introduced.  
     
     
         77 . The cell of  claim 76  wherein said hybrid cell is a dendritic cell/tumor cell hybrid.  
     
     
         78 . The cells of claims  73 - 77  wherein said mammal bears a tumor, against which the antitumor response is directed, selected from the group consisting of a carcinoma, a melanoma, a sarcoma, a neuroblastoma, a lymphoma and a leukemia.  
     
     
         79 . The cell of claim  73 - 77  wherein the superantigen is selected from the group consisting of a Staphylococcal enterotoxin and a Streptococcal pyrogenic exotoxin.  
     
     
         80 . A method of treating a tumor or neoplastic disease in a subject comprising administering an effective amount of nucleic acids in vivo to the tumor/accessory tumor/accessory cell hybrid cells of claims  76 - 77 .  
     
     
         81 . A method of treating a tumor or neoplastic disease in a subject comprising administering an effective amount of the cells of claim  76 - 77  wherein said exogenous nucleic acids are introduced into the cell ex vivo or in vitro.