Melanoma vaccine and methods of making and using same

Abstract

An immunotherapeutic vaccine providing antigen presenting cells that have been pulsed with a disrupted cell preparation which includes enucleated cytosol and cell membranes of cancer cells infected with a recombinant vaccinia virus encoding at least one immunostimulating molecule. In a preferred embodiment, the vaccine includes autologous dendritic/monocytic cells (DC/M) that present a mixture of antigens (present in the enucleated cytosol and cell membranes) from melanoma cell lines that have been infected with a recombinant vaccinia virus encoding IL-2. In another of the preferred embodiments, the enucleated cytosol and cell membranes are from melanoma cells harvested from the patient to be treated. A method of making the vaccine and methods of using the vaccine to stimulate an anti-cancer immune response and to treat a patient with a cancer are also described.

1 - 17 . (canceled)  
     
     
         18 . An immunotherapeutic vaccine comprising: 
 (a) a first part comprising a first recombinant vaccinia virus encoding at least one first immunostimulating molecule, wherein the first immunostimulating molecule is IL-2; and    (b) a second part comprising antigen presenting cells, which are capable of inducing T-cell activation, wherein the antigen presenting cells are dendritic cells and/or a monocytes, and which are autologous or syngeneic, pulsed with a preparation comprising enucleated cytosol and cell membranes of cancer cells infected with a second recombinant vaccinia virus encoding at least one second immunostimulating molecule, wherein the second immunostimulating molecule is IL-2.    
     
     
         19 . The vaccine of  claim 18 , wherein the enucleated cytosol is substantially free of nuclei.  
     
     
         20 . The vaccine of  claim 18 , wherein the cell membranes comprise at least two HLA class I A antigens.  
     
     
         21 . The vaccine of  claim 18 , wherein the first recombinant vaccinia virus is a live virus.  
     
     
         22 . The vaccine of  claim 18 , wherein the second recombinant vaccinia virus is either live or inactivated.  
     
     
         23 - 28 . (canceled)  
     
     
         29 . The vaccine of  claim 18 , wherein the antigen presenting cells are dendritic cells or monocytes.  
     
     
         30 . The vaccine of  claim 18 , wherein the antigen presenting cells are dendritic cells and monocytes.  
     
     
         31 . The vaccine of  claim 18 , wherein the antigen presenting cells are autologous cells.  
     
     
         32 . The vaccine of  claim 18 , wherein the antigen presenting cells are HLA-matched to a host to be treated.  
     
     
         33 . The vaccine of  claim 18 , wherein the cancer cells are melanoma cells.  
     
     
         34 . The vaccine of  claim 33 , wherein the melanoma cells comprise one or more cells selected from the group consisting of Mel-2, Mel-3, Mel-4, Mel-6, and Mel-9.  
     
     
         35 . The vaccine of  claim 18 , wherein the cancer cells are established cancer cell lines.  
     
     
         36 . The vaccine of  claim 18 , wherein the cancer cells are selected from the group consisting of fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, Kaposi's sarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, rhabdosarcoma, colorectal carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, neuroblastoma, retinoblastoma, myeloma, lymphoma, and leukemia cells.  
     
     
         37 . The vaccine of  claim 35 , wherein the cancer cell lines are selected from the group of cancer cell lines consisting of fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, Kaposi's sarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, rhabdosarcoma, colorectal carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, neuroblastoma, retinoblastoma, myeloma, lymphoma, and leukemia cell lines.  
     
     
         38 . The vaccine of  claim 18 , wherein the cancer cells are harvested from a host to be treated with the vaccine.  
     
     
         39 - 107 . (canceled)  
     
     
         108 . A method for eliciting an anti-cancer immune response in a subject, which comprises: 
 (a) administering a first recombinant vaccinia virus encoding at least one first immunostimulating molecule, wherein the first immunostimulating molecule is IL-2; and    (b) administering a composition comprising antigen presenting cells, which are capable of inducing T cell activation, wherein the antigen presenting cells are dendritic cells and/or monocytes, and which are autologous or syngeneic, pulsed with a preparation comprising enucleated cytosol and cell membranes of cancer cells infected with a second recombinant vaccinia virus encoding at least one second immunostimulating molecule, wherein the second immunostimulating molecule is IL-2.    
     
     
         109 . The method of  claim 108 , wherein about 10 4  to 10 8  PFU of the first recombinant vaccinia virus are provided.  
     
     
         110 . The method of  claim 108 , wherein about 10 7  PFU of the first recombinant vaccinia virus are provided.  
     
     
         111 . The method of  claim 108 , wherein about 10 5  to 10 7  antigen presenting cells are provided.  
     
     
         112 . The method of  claim 108 , wherein about 10 6  to 5×10 6  antigen presenting cells are provided.  
     
     
         113 . The method of  claim 108 , wherein the antigen presenting cells are dendritic cells or monocytes.  
     
     
         114 . The method of  claim 108 , wherein the antigen presenting cells are dendritic cells and monocytes.  
     
     
         115 . The method of  claim 108 , wherein the antigen presenting cells are autologous cells.  
     
     
         116 . The method of  claim 108 , wherein the antigen presenting cells are HLA-matched to the subject.  
     
     
         117 . The method of  claim 108 , wherein the cancer cells are melanoma cells.  
     
     
         118 . The method of  claim 117 , wherein the melanoma cells comprise one or more cells selected from the group consisting of Mel-2, Mel-3, Mel-4, Mel-6, and Mel-9.  
     
     
         119 . The method of  claim 108 , wherein the cancer cells are from established cancer cell lines.  
     
     
         120 . The method of  claim 108 , wherein the cancer cells are harvested from the subject.