Method of using a cytokine that induces apoptosis

Abstract

A novel cytokine designated TRAIL induces apoptosis of certain target cells, including cancer cells and virally infected cells. Isolated DNA sequences encoding TRAIL are disclosed, along with expression vectors and transformed host cells useful in producing TRAIL polypeptides. Antibodies that specifically bind TRAIL are provided as well.

1 - 28 . (canceled)  
     
     
         29 . A method of inducing death of cancer cells, comprising contacting TRAIL-sensitive cancer cells with TRAIL polypeptide, wherein said polypeptide comprises an amino acid sequence that is at least 90% identical to an amino acid sequence selected from the group consisting of amino acids 1 to 281 of SEQ ID NO:2 and amino acids 1 to 291 of SEQ ID NO:6, wherein said TRAIL polypeptide is capable of inducing apoptosis of Jurkat cells.  
     
     
         30 . A method of inducing death of cancer cells, comprising contracting TRAIL-sensitive cancer cells with an oligomer comprising at least two soluble TRAIL polypeptides, wherein each of said soluble TRAIL polypeptides comprises an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of: 
 a) the extracellular domain of human TRAIL (amino acids 39 to 281 of SEQ ID NO:2); and    b) a fragment of said extracellular domain;    wherein said oligomer is capable of inducing apoptosis of Jurkat cells.    
     
     
         31 . A method of  claim 29 , wherein said TRAIL polypeptide comprises an amino acid sequence selected from the group consisting of amino acids 1 to 281 of SEQ ID NO:2 and amino acids 1 to 291 of SEQ ID NO:6.  
     
     
         32 . A method of inducing death of cancer cells, comprising contacting TRAIL-sensitive cancer cells with a soluble polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of: 
 a) the extracellular domain of human TRAIL (amino acids 39 to 281 of SEQ ID NO:2); and    b) a fragment of said extracellular domain;    wherein said polypeptide is capable of inducing apoptosis of Jurkat cells.    
     
     
         33 . A method of  claim 32 , wherein said polypeptide comprises an amino acid sequence selected from the group consisting of: 
 a) the extracellular domain of human TRAIL (amino acids 39 to 281 of SEQ ID NO:2); and    b) a fragment of said extracellular domain;    wherein said polypeptide is capable of inducing apoptosis of Jurkat cells.    
     
     
         34 . A method of  claim 32 , wherein said polypeptide comprises an amino acid sequence that is at least 90% identical to the sequence of amino acids 95 to 281 of SEQ ID NO:2, wherein said polypeptide is capable of inducing apoptosis of Jurkat cells.  
     
     
         35 . A method of  claim 33 , wherein said polypeptide comprises the sequence of amino acids x to 281 of SEQ ID NO:2, wherein x represents an integer from 39 to 95.  
     
     
         36 . A method of  claim 35 , wherein said polypeptide comprises amino acids 95 to 281 of SEQ ID NO:2.  
     
     
         37 . A method of inducing death of cancer cells, comprising contacting TRAIL-sensitive cancer cells with a polypeptide that comprises a fragment of the human TRAIL protein of SEQ ID NO:2, wherein said fragment kills TRAIL-sensitive cancer cells.  
     
     
         38 . A method of  claim 37 , wherein said polypeptide consists of a soluble fragment of the human TRAIL protein of SEQ ID NO:2.  
     
     
         39 . A method of inducing death of cancer cells, comprising contacting TRAIL-sensitive cancer cells with a polypeptide comprising amino acids 124 to 276 of SEQ ID NO:2.  
     
     
         40 . A method of  claim 39 , wherein said polypeptide is a fragment of the TRAIL protein of SEQ ID NO:2, wherein the N-terminal amino acid of said fragment is selected from residues 39 to 124 of SEQ ID NO:2, and the C-terminal amino acid of said fragment is selected from residues 276 to 281 of SEQ ID NO:2.  
     
     
         41 . A method of  claim 37 , wherein said polypeptide comprises the amino acid sequence presented in SEQ ID NO:2 or SEQ ID NO:6, with the proviso that said polypeptide lacks a transmembrane region.  
     
     
         42 . A method of  claim 32 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         43 . A method of  claim 33 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         44 . A method of  claim 37 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         45 . A method of  claim 39 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         46 . A method of  claim 40 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         47 . A method of  claim 30 , wherein each of said soluble TRAIL polypeptides comprises an amino acid sequence selected from the group consisting of: 
 a) the extracellular domain of human TRAIL (amino acids 39 to 281 of SEQ ID NO:2); and    b) a fragment of said extracellular domain;    wherein said oligomer is capable of inducing apoptosis of Jurkat cells.    
     
     
         48 . A method of  claim 30 , wherein each of said soluble TRAIL polypeptides comprises an amino acid sequence that is at least 90% identical to the sequence of amino acids 95 to 281 of SEQ ID NO:2, wherein said polypeptide induces apoptosis of Jurkat cells.  
     
     
         49 . A method of  claim 47 , wherein each of said soluble TRAIL polypeptides comprises the sequence of amino acids x to 281 of SEQ ID NO:2, wherein x represents an integer from 39 to 95.  
     
     
         50 . A method of  claim 49 , wherein each of said soluble TRAIL polypeptides comprises amino acids 95 to 281 of SEQ ID NO:2.  
     
     
         51 . A method of killing cancer cells, contacting TRAIL-sensitive cancer cells with an oligomer comprising at least two TRAIL polypeptides, wherein each of said TRAIL polypeptides is a soluble fragment of the protein of SEQ ID NO:2, wherein said oligomer kills TRAIL-sensitive cancer cells.  
     
     
         52 . A method of inducing death of cancer cells, comprising contacting TRAIL-sensitive cancer cells with an oligomer comprising at least two TRAIL polypeptides, wherein each of said polypeptides comprises amino acids 124 to 276 of SEQ ID NO:2.  
     
     
         53 . A method of  claim 52 , wherein each of said polypeptides is a fragment of the TRAIL protein of SEQ ID NO:2, wherein the N-terminal amino acid of said fragment is selected from residues 39 to 124 of SEQ ID NO:2, and the C-terminal amino acid of said fragment is selected from residues 276 to 281 of SEQ ID NO:2.  
     
     
         54 . A method of  claim 30 , wherein each of said polypeptides additionally comprises a leucine zipper peptide.  
     
     
         55 . A method of  claim 47 , wherein each of said polypeptides additionally comprises a leucine zipper peptide.  
     
     
         56 . A method of  claim 30 , wherein said oligomer additionally comprises an Fc polypeptide fused to each of two of said TRAIL polypeptides.  
     
     
         57 . A method of  claim 47 , wherein said oligomer additionally comprises an Fc polypeptide fused to each of two of said TRAIL polypeptides.  
     
     
         58 . A method of  claim 51 , wherein said oligomer additionally comprises an Fc polypeptide fused to each of two of said TRAIL polypeptides.  
     
     
         59 . A method of  claim 30 , wherein the oligomer comprises two or three of said TRAIL polypeptides.  
     
     
         60 . A method of  claim 47 , wherein the oligomer comprises two or three of said TRAIL polypeptides.  
     
     
         61 . A method of  claim 59 , wherein the oligomer comprises three of said TRAIL polypeptides.  
     
     
         62 . A method of  claim 60 , wherein the oligomer comprises three of said TRAIL polypeptides.  
     
     
         63 . A method of  claim 51 , wherein the oligomer comprises three of said TRAIL polypeptides.  
     
     
         64 . A method of  claim 52 , wherein the oligomer comprises three of said TRAIL polypeptides.  
     
     
         65 . A method of  claim 53 , wherein the oligomer comprises three of said TRAIL polypeptides.  
     
     
         66 . A method of  claim 30 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         67 . A method of  claim 47 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         68 . A method of  claim 51 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         69 . A method of  claim 52 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         70 . A method of  claim 53 , wherein said cancer cells are selected from the group consisting of leukemia cells, melanoma cells, and lymphoma cells.  
     
     
         71 . A method of  claim 30 , wherein the method comprises administering said oligomer to a human who has cancer.  
     
     
         72 . A method of  claim 32 , wherein the method comprises administering said polypeptide to a human who has cancer.  
     
     
         73 . A method of  claim 37 , wherein the method comprises administering said polypeptide to a human who has cancer.  
     
     
         74 . A method of  claim 39 , wherein the method comprises administering said polypeptide to a human who has cancer.  
     
     
         75 . A method of  claim 47 , wherein the method comprises administering said oligomer to a human who has cancer.  
     
     
         76 . A method of  claim 51 , wherein the method comprises administering said oligomer to a human who has cancer.  
     
     
         77 . A method of  claim 52 , wherein the method comprises administering said oligomer to a human who has cancer.  
     
     
         78 . A method of  claim 53 , wherein the method comprises administering said oligomer to a human who has cancer.  
     
     
         79 . A method of treating a mammal having cancer, comprising administering to the mammal a soluble, extracellular TRAIL polypeptide in an amount effective to induce cell death in the mammal's cancer cells, wherein the TRAIL polypeptide comprises a soluble fragment of the TRAIL protein of SEQ ID NO:2, wherein the N-terminal amino acid of said fragment is selected from residues 39 to 124 of SEQ ID NO:2, and the C-terminal amino acid of said fragment is selected from residues 276 to 281 of SEQ ID NO:2.  
     
     
         80 . A method of  claim 79 , wherein said mammal is a human, and said TRAIL polypeptide consists of a soluble fragment of the TRAIL protein of SEQ ID NO:2, wherein the N-terminal amino acid of said fragment is selected from residues 39 to 124 of SEQ ID NO:2, and the C-terminal amino acid of said fragment is selected from residues 276 to 281 of SEQ ID NO:2.  
     
     
         81 . A method of  claim 79 , wherein said method additionally comprises administering another agent that exerts a cytotoxic effect on the cancer cells.  
     
     
         82 . A method of  claim 80 , wherein said method additionally comprises administering another agent that exerts a cytotoxic effect on the cancer cells.  
     
     
         83 . A method of  claim 79 , wherein said polypeptide is in the form of a composition comprising the polypeptide and a physiologically acceptable carrier, diluent, or excipient.  
     
     
         84 . A method of treating a mammal having cancer, comprising administering to the mammal a soluble, extracellular TRAIL polypeptide in an amount effective to induce cell death in the mammal's cancer cells, wherein the TRAIL polypeptide comprises amino acids 124 to 281 of SEQ ID NO:2.  
     
     
         85 . A method of  claim 84 , wherein said mammal is a human.  
     
     
         86 . A method comprising administering an oligomer to a human who has cancer, wherein said oligomer comprises at least two TRAIL polypeptides, wherein each of said polypeptides is a soluble fragment of the protein of SEQ ID NO:2, wherein said oligomer kills TRAIL-sensitive cancer cells.  
     
     
         87 . A method of  claim 86 , wherein the N-terminal amino acid of said fragment is selected from residues 39 to 124 of SEQ ID NO:2, and the C-terminal amino acid of said fragment is selected from residues 276 to 281 of SEQ ID NO:2.  
     
     
         88 . A method of  claim 86 , wherein each of said polypeptides comprises amino acids 124 to 276 of SEQ ID NO:2.  
     
     
         89 . A method of  claim 86 , wherein the oligomer comprises two or three of said TRAIL polypeptides.  
     
     
         90 . A method of  claim 87 , wherein the oligomer comprises three of said TRAIL polypeptides.  
     
     
         91 . A method of killing target cells selected from the group consisting of cancer cells and virally-infected cells, comprising contacting the target cells with an oligomer comprising at least two TRAIL polypeptides, wherein each of said TRAIL polypeptides is a soluble fragment of the protein of SEQ ID NO:2, wherein said oligomer kills TRAIL-sensitive target cells.  
     
     
         92 . A method of  claim 91 , wherein the N-terminal amino acid of said fragment is selected from residues 39 to 124 of SEQ ID NO:2, and the C-terminal amino acid of said fragment is selected from residues 276 to 281 of SEQ ID NO:2.  
     
     
         93 . A method of  claim 91 , wherein each of said polypeptides comprises amino acids 124 to 276 of SEQ ID NO:2.  
     
     
         94 . A method of  claim 92 , wherein the oligomer comprises three of said TRAIL polypeptides.  
     
     
         95 . A method of  claim 91 , wherein said method additionally comprises administering another agent that exerts a cytotoxic effect on the target cells.  
     
     
         96 . A method of  claim 86 , wherein said oligomer is in the form of a composition comprising the oligomer and a physiologically acceptable carrier, diluent, or excipient.  
     
     
         97 . A method of  claim 91 , wherein said oligomer is in the form of a composition comprising the oligomer and a physiologically acceptable carrier, diluent, or excipient.  
     
     
         98 . A method of inducing death of cancer cells in vitro, comprising contacting TRAIL-sensitive cancer cells with a soluble polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence selected from the group consisting of: 
 a) the extracellular domain of human TRAIL (amino acids 39 to 281 of SEQ ID NO:2); and    b) a fragment of said extracellular domain;    wherein said polypeptide is capable of inducing apoptosis of Jurkat cells.    
     
     
         99 . The method of  claim 98 , wherein said polypeptide comprises an amino acid sequence selected from the group consisting of: 
 a) the extracellular domain of human TRAIL (amino acids 39 to 281 of SEQ ID NO:2); and    b) a fragment of said extracellular domain;    wherein said polypeptide is capable of inducing apoptosis of Jurkat cells.    
     
     
         100 . A method of killing target cells in vitro wherein the target cells are cancer cells or virally-infected cells, comprising contacting the target cells with an oligomer comprising at least two TRAIL polypeptides, wherein each of said TRAIL polypeptides is a soluble fragment of the protein of SEQ ID NO:2, wherein said oligomer kills TRAIL-sensitive target cells.  
     
     
         101 . The method of  claim 100 , wherein the N-terminal amino acid of said fragment is selected from residues 39 to 124 of SEQ ID NO:2, and the C-terminal amino acid of said fragment is selected from residues 276 to 281 of SEQ ID NO:2.