Tumor cells with increased immunogenicity and uses therefor

Abstract

Tumor cells modified to express a T cell costimulatory molecule are disclosed. In one embodiment, the costimulatory molecule is a CD28/CTLA4 ligand, preferably a B lymphocyte antigen B7. The tumor cells of the invention can be modified by transfection with nucleic acid encoding a T cell costimulatory molecule, by using an agent which induces or increases expression of a T cell costimulatory molecule on the tumor cell surface or by coupling a T cell costimulatory molecule to the tumor cell surface. Tumor cells further modified to express MHC class I and/or class II molecules or in which expression of an MHC associated protein, the invariant chain, is inhibited are also disclosed. The modified tumor cells of the invention can be used in methods for treating a patient with a tumor, preventing or inhibiting metastatic spread of a tumor or preventing or inhibiting recurrence of a tumor. A method for specifically inducing a CD4+ T cell response against a tumor and a method for treating a tumor by modification of tumor cells in vivo are disclosed.

1 . A tumor cell which is modified to express a T cell costimulatory molecule.  
     
     
         2 . The tumor cell of  claim 1  which is transfected with a nucleic acid encoding a T cell costimulatory molecule in a form suitable for expression of the costimulatory molecule.  
     
     
         3 . The tumor cell of  claim 2  wherein the T cell costimulatory molecule is a CD28 and/or CTLA4 ligand.  
     
     
         4 . The tumor cell of  claim 3  wherein the CD28 and/or CTLA4 ligand is a B lymphocyte antigen, B7.  
     
     
         5 . The tumor cell of  claim 1  which is stimulated to express a T cell costimulatory molecule.  
     
     
         6 . The tumor cell of  claim 5  wherein the T cell costimulatory molecule is a CD28 and/or CTLA4 ligand.  
     
     
         7 . The tumor cell of  claim 6  wherein the CD28 and/or CTLA4 ligand is a B lymphocyte antigen, B7.  
     
     
         8 . The tumor cell of  claim 1  which has a T cell costimulatory molecule coupled to the tumor cell.  
     
     
         9 . The tumor cell of  claim 8  wherein the T cell costimulatory molecule is a CD28 and/or CTLA4 ligand.  
     
     
         10 . The tumor cell of  claim 9  wherein the CD28 and/or CTLA4 ligand is a B lymphocyte antigen, B7.  
     
     
         11 . The tumor cell of  claim 1  which expresses an MHC class II molecule.  
     
     
         12 . The tumor cell of  claim 1  which expresses an MHC class I molecule.  
     
     
         13 . The tumor cell of  claim 1  which normally expresses an MHC class II associated protein, the invariant chain, and wherein expression of the invariant chain is inhibited.  
     
     
         14 . A tumor cell transfected with a nucleic acid encoding a B lymphocyte antigen, B7, in a form suitable for expression of B7.  
     
     
         15 . The tumor cell of  claim 14  wherein the nucleic acid is a cDNA in a recombinant expression vector.  
     
     
         16 . The tumor cell of  claim 15  further transfected with at least one nucleic acid comprising DNA encoding: 
 (a) at least one MHC class II α chain protein; and  
 (b) at least one MHC class II β chain protein,  
 wherein the nucleic acid is in a form suitable for expression of the MHC class II α chain protein(s) and the MHC class II β chain protein(s).  
 
     
     
         17 . The tumor cell of  claim 16  which does not express MHC class II molecules prior to transfection of the tumor cell.  
     
     
         18 . The tumor cell of  claim 14  further transfected with at least one nucleic acid encoding at least one MHC class I α chain protein in a form suitable for expression of the MHC class I protein(s).  
     
     
         19 . The tumor cell of  claim 18  further transfected with a nucleic acid encoding a β-2 microglobulin protein in a form suitable for expression of the β-2 microglobulin protein.  
     
     
         20 . The tumor cell of  claim 14  which normally expresses an MHC class II associated protein, the invariant chain, and wherein expression of the invariant chain is inhibited.  
     
     
         21 . The tumor cell of  claim 20  wherein expression of the invariant chain is inhibited by transfection of the tumor cell with a nucleic acid which is antisense to a regulatory or a coding region of the invariant chain gene.  
     
     
         22 . The tumor cell of  claim 14  which is a sarcoma.  
     
     
         23 . The tumor cell of  claim 14  which is a lymphoma.  
     
     
         24 . The tumor cell of  claim 14  which is selected from a group consisting of a melanoma, a neuroblastoma, a leukemia and a carcinoma.  
     
     
         25 . A sarcoma cell which is modified to express a T cell costimulatory molecule.  
     
     
         26 . The sarcoma cell of  claim 25  which is transfected with a nucleic acid encoding a T cell costimulatory molecule in a form suitable for expression of the costimulatory molecule.  
     
     
         27 . The sarcoma cell of  claim 26  wherein the T cell costimulatory molecule is a CD28 and/or CTLA4 ligand.  
     
     
         28 . The sarcoma cell of  claim 27  wherein the CD28 and/or CTLA4 ligand is a B lymphocyte antigen, B7.  
     
     
         29 . The sarcoma cell of  claim 25  which expresses an MHC class II molecule.  
     
     
         30 . The sarcoma cell of  claim 25  which expresses an MHC class I molecule.  
     
     
         31 . A lymphoma cell transfected with a nucleic acid encoding a B lymphocyte antigen, B7, in a form suitable for expression of B7.  
     
     
         32 . The lymphoma cell of  claim 31  wherein the nucleic acid is a cDNA in a recombinant expression vector.  
     
     
         33 . A composition suitable for pharmaceutical administration comprising an amount of the tumor cells of  claim 1  and a physiologically acceptable carrier.  
     
     
         34 . A composition suitable for pharmaceutical administration comprising an amount of the tumor cells of  claim 14  and a physiologically acceptable carrier.  
     
     
         35 . A composition suitable for inducing an anti-tumor response by CD4+ T helper lymphocytes in a subject with a tumor comprising an amount of tumor cells of  claim 16  and a physiologically acceptable carrier.  
     
     
         36 . A composition suitable for pharmaceutical administration comprising an amount of the sarcoma cells of  claim 25  and a physiologically acceptable carrier.  
     
     
         37 . A composition suitable for pharmaceutical administration comprising an amount of the lymphoma cells of  claim 31  and a physiologically acceptable carrier.  
     
     
         38 . A method for treating a subject with a tumor, comprising: 
 (a) obtaining tumor cells from the subject;    (b) modifying the tumor cells to express a T cell costimulatory molecule; and    (c) administering the tumor cells to the subject.    
     
     
         39 . The method of  claim 38  wherein tumor cells are modified by transfection with a nucleic acid encoding a T cell costimulatory molecule in a form suitable for expression of the T cell costimulatory molecule.  
     
     
         40 . The method of  claim 38  wherein tumor cells are modified by treatment with an agent which stimulates expression of a T cell costimulatory molecule.  
     
     
         41 . The method of  claim 38  wherein tumor cells are modified by coupling a T cell costimulatory molecule to the tumor cell.  
     
     
         42 . A method of treating a subject with a tumor, comprising: 
 (a) obtaining tumor cells from the subject;    (b) transfecting the tumor cells with a nucleic acid encoding a CD28 and/or CTLA-4 ligand in a form suitable for expression of the CD28 and/or CTLA4 ligand; and    (c) administering the tumor cells to the subject.    
     
     
         43 . The method of  claim 42  wherein the CD28 and/or CTLA-4 ligand is a B lymphocyte antigen, B7.  
     
     
         44 . The method of  claim 42  wherein the tumor cells are further transfected with at least one nucleic acid encoding at least one MHC class II α chain protein and at least one MHC class II β chain protein in a form suitable for expression of the MHC class II α chain protein(s) and the MHC class II β chain protein(s).  
     
     
         45 . The method of  claim 42  wherein the tumor cells are further transfected with at least one nucleic acid encoding at least one MHC class I α chain protein in a form suitable for expression of the MHC class I protein(s).  
     
     
         46 . The method of  claim 45  wherein the tumor cells are further transfected with a nucleic acid encoding a β-2 microglobulin protein in a form suitable for expression of the β-2 microglobulin protein.  
     
     
         47 . The method of  claim 42  wherein expression of an MHC class II associated protein, the invariant chain, is inhibited in the tumor cells.  
     
     
         48 . The method of  claim 47  wherein expression of the invariant chain is inhibited in the tumor cells by transfection of the tumor cell with a nucleic acid which is antisense to a regulatory or a coding region of the invariant chain gene.  
     
     
         49 . The method of  claim 47  wherein the tumor is a sarcoma.  
     
     
         50 . The method of  claim 47  wherein the tumor is a lymphoma.  
     
     
         51 . The method of  claim 47  wherein the tumor is selected from a group consisting of a melanoma, a neuroblastoma, a leukemia and a carcinoma.  
     
     
         52 . The method of  claim 47  wherein the tumor cells are administered by intravenous injection.  
     
     
         53 . The method of  claim 47  wherein the tumor cells are administered by a route selected from a group consisting of intramuscular injection, intraperitoneal injection and subcutaneous injection.  
     
     
         54 . A method for preventing or treating metastatic spread of a tumor or preventing or treating recurrence of a tumor in a subject, comprising: 
 (a) obtaining tumor cells from the subject;    (b) transfecting the tumor cells with a nucleic acid encoding a CD28 and/or CTLA-4 ligand in a form suitable for expression of the CD28 and/or CTLA-4 ligand; and    (c) administering the tumor cells to the subject.    
     
     
         55 . The method of  claim 54  wherein the CD28 and/or CTLA-4 ligand is a B lymphocyte antigen, B7.  
     
     
         56 . The method of  claim 54  wherein the tumor is a sarcoma.  
     
     
         57 . The method of  claim 54  wherein the tumor is a lymphoma.  
     
     
         58 . The method of  claim 54  wherein the tumor is selected from a group consisting of a melanoma, a neuroblastoma, a leukemia and a carcinoma.  
     
     
         59 . The method of  claim 54  wherein the tumor cells are administered by intravenous injection.  
     
     
         60 . The method of  claim 54  wherein the tumor cells are administered by a route selected from a group consisting of intramuscular injection, intraperitoneal injection and subcutaneous injection.  
     
     
         61 . A method of inducing an anti-tumor response by CD4+ T lymphocytes in a subject with a tumor, comprising: 
 (a) obtaining tumor cells from the subject;    (b) transfecting the tumor cells with at least one nucleic acid comprising DNA encoding a B lymphocyte antigen, B7, an MHC class II α chain protein and an MHC class II β chain protein, wherein the nucleic acid is in a form suitable for expression of B7, the MHC class II α chain protein and the MHC class II β chain protein; and    (c) administering the tumor cells to the subject.    
     
     
         62 . The method of  claim 61  wherein the tumor is a sarcoma.  
     
     
         63 . The method of  claim 61  wherein the tumor is a lymphoma.  
     
     
         64 . The method of  claim 61  wherein the tumor is selected from a group consisting of a melanoma, a neuroblastoma, a leukemia and a carcinoma.  
     
     
         65 . The method of  claim 61  wherein the tumor cells are administered by intravenous injection.  
     
     
         66 . The method of  claim 61  wherein the tumor cells are administered by a route selected from a group consisting of intramuscular injection, intraperitoneal injection and subcutaneous injection.  
     
     
         67 . A method for treating a subject with a tumor comprising modifying tumor cells in vivo to express a T cell costimulatory molecule.  
     
     
         68 . The method of  claim 67  wherein the T cell costimulatory molecule is a CD28 and/or CTLA4 ligand.  
     
     
         69 . The method of  claim 68  wherein the CD28 and/or CTLA4 ligand is a B lymphocyte antigen, B7.  
     
     
         70 . The method of  claim 67  wherein tumor cells are modified in vivo by delivering to the subject in vivo a nucleic acid encoding a T cell costimulatory molecule in a form suitable for expression of the T cell costimulatory molecule.  
     
     
         71 . The method of  claim 70  wherein the nucleic acid is delivered to the subject in vivo by injection of the nucleic acid in an appropriate vehicle into the tumor.  
     
     
         72 . A method for treating a subject with a tumor, comprising: 
 (a) obtaining tumor cells and T lymphocytes from the subject;    (b) culturing the T lymphocytes from the subject in vitro with the tumor cells from the subject and with a stimulatory form of a T cell costimulatory molecule; and    (c) administering the T lymphocytes to the subject.