Antigen presenting system and methods for activation of T-cells

Abstract

Materials and methods for activating T lymphocytes with specificity for particular antigenic peptides are described, as well as the use of activated T lymphocytes in vitro for the treatment of a variety of disease conditions. In particular, a method for producing a synthetic antigen presenting cell line for activating T lymphocytes to a specific peptide is described.

We claim:  
     
         1 . A synthetic antigen presenting cell line for activating T-cell lymphocytes to a specific peptide and comprising: 
 a) a Class I MHC heavy chain gene operably linked to a first promoter, said gene expressing a Class I MHC heavy chain;    b) a β-2 microglobulin gene operably linked to a second promoter, said gene expressing a β-2 microglobulin that forms a MHC molecule with the MHC heavy chain; and    c) a gene for an assisting molecule operably linked to a third promoter, said gene expressing an assisting molecule which is a member of the group consisting of B7.1, B7.2, ICAM-1, ICAM-2, ICAM-3 and LFA-3, and that interacts with a molecule on the T-cell lymphocytes;    with the proviso that at least one of said MHC gene, said β-2 microglobulin gene and said assisting molecule gene is not present in the cells from which the cell line is derived, such that the MHC molecule binds to a peptide, and the MHC molecule and the assisting molecule are presented on the surface of an insect cell of said cell line in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the MHC molecule.    
     
     
         2 . The cell line of  claim 1  wherein the assisting molecule is a costimulatory molecule.  
     
     
         3 . The cell line of  claim 1  wherein the cell line is derived from a first species and the MHC heavy chain gene is from a second species.  
     
     
         4 . The cell line of  claim 3  wherein the first species is a poikilotherm and the second species is a homeotherm.  
     
     
         5 . The cell line of  claim 1  wherein the assisting molecule is an adhesion molecule.  
     
     
         6 . The cell line of  claim 5  wherein the adhesion molecule is a member of the group consisting of ICAM-1, ICAM-2, ICAM-3 and LFA-3.  
     
     
         7 . The cell line of  claim 1  having a gene for a first assisting molecule and a gene for a second assisting molecule.  
     
     
         8 . The cell line of  claim 7  wherein the first assisting molecule is a costimulatory molecule and the second assisting molecule is an adhesion molecule.  
     
     
         9 . The cell line of  claim 1  wherein at least one promoter is inducible.  
     
     
         10 . The cell line of  claim 9  wherein the first promoter is inducible.  
     
     
         11 . The cell line of  claim 1  wherein the peptide is bound to the MHC molecule within the cell.  
     
     
         12 . The cell line of  claim 1  wherein the MHC molecule is presented empty on the surface of the cell.  
     
     
         13 . A stable poikilotherm cell line used to stimulate human T-cell lymphocytes comprising: 
 a) a Class I MHC gene operably linked to a first inducible promoter, said gene expressing a Class I MHC heavy chain;    b) a β-2 microglobulin gene operably linked to a second promoter, said gene expressing a β-2 microglobulin that forms a MHC molecule with the MHC heavy chain; and    c) a gene for an assisting molecule operably linked to a third promoter, said gene expressing an assisting molecule which is a member of the group consisting of B7.1, B7.2, ICAM-1, ICAM-2, ICAM-3 and LFA-3, and that interacts with a molecule on the T-cell lymphocytes;    such that a cell of the cell line assembles empty MHC molecules that bind to a peptide, and the MHC molecule and the assisting molecule are presented on the surface of a cell of the cell line in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the MHC molecule.    
     
     
         14 . The cell line of  claim 13  wherein the assisting molecule is a costimulatory molecule.  
     
     
         15 . The cell line of  claim 13  wherein the assisting molecule is an adhesion molecule.  
     
     
         16 . The cell line of  claim 15  wherein the adhesion molecule is ICAM-1.  
     
     
         17 . The cell line of  claim 13  having a gene for a first assisting molecule and a gene for a second assisting molecule.  
     
     
         18 . The cell line of  claim 17  wherein the first assisting molecule is a costimulatory molecule and the second assisting molecule is an adhesion molecule.  
     
     
         19 . A stable poikilotherm cell line used to stimulate human T-cell lymphocytes comprising: 
 a) a Class I MHC gene operably linked to a first promoter, said gene expressing a Class I MHC heavy chain;    b) a β-2 microglobulin gene operably linked to a second promoter, said gene expressing a β-2 microglobulin that forms a MHC molecule with the MHC heavy chain; and    c) a gene for a costimulatory molecule operably linked to a third promoter and said gene expressing a costimulatory molecule which is a B7.1 molecule or a B7.2 molecule that interacts with a molecule on the T-cell lymphocytes;    d) a gene for an adhesion molecule operably linked to a fourth promoter and said gene expressing an adhesion molecule that interacts with a cooperative adhesion molecule on the T-cell lymphocytes and is a member of the group consisting of ICAM-1, ICAM-2, ICAM-3 and LFA-3.    such that the cell is capable of assembling the MHC heavy chain and the β-2 microglobulin into a MHC molecule that binds to a peptide, and transporting MHC molecule, costimulatory molecule and adhesion molecule to the surface of the cell in sufficient numbers to activate a population of T-cell lymphocytes against the peptide when the peptide is bound to the MHC molecule.    
     
     
         20 . The cell line of  claim 19  wherein at least one of the promoters is inducible.  
     
     
         21 . The cell line of  claim 19  wherein the peptide is bound to the MHC molecule within the cell.  
     
     
         22 . The cell line of  claim 19  wherein the MHC molecule is presented empty on the surface of the cell.