Compositions comprising a first agent which provides a primary activation signal to T cells and a second agent which stimulates an accessory molecule on the surface of T cells

Abstract

Compositions for inducing a population of T cells to proliferate in vitro by activating the population of T cells and stimulating an accessory molecule on the surface of the T cells with a ligand which binds the accessory molecule are described. T cell proliferation occurs in the absence of exogenous growth factors or accessory cells. T cell activation is accomplished by stimulating the T cell receptor (TCR)/CD3 complex or the CD2 surface protein. To induce proliferation of an activated population of T cells, an accessory molecule on the surface of the T cells, such as CD28, is stimulated with a ligand which binds the accessory molecule. The T cell population expanded by the composition of the invention can be genetically transduced and used for immunotherapy or can be used in methods of diagnosis

1 . A composition comprising a magnetic bead having covalently attached thereto: 
 (a) an anti-CD3 antibody or a CD3-binding fragment thereof; and    (b) an anti-CD28 antibody or a CD28-binding fragment thereof, wherein the anti-CD3 antibody or CD3-binding fragment thereof and the anti-CD28 antibody or CD28-binding fragment thereof are covalently attached to the same magnetic bead.    
     
     
         2 . The composition of  claim 1 , wherein the anti-CD3 antibody or a CD3-binding fragment thereof is an anti-human CD3 monoclonal antibody or a CD3-binding fragment thereof.  
     
     
         3 . The composition of  claim 1 , wherein the anti-CD28 antibody or a CD28-binding fragment thereof is an anti-human CD28 monoclonal antibody or a CD28-binding fragment thereof.  
     
     
         4 . The composition of  claim 1 , wherein the anti-CD3 antibody or CD3-binding fragment thereof and the anti-CD28 antibody or CD28-binding fragment thereof are covalently attached to the magnetic bead via a tosyl linkage.  
     
     
         5 . The composition of  claim 1 , wherein the anti-CD3 antibody is OKT3.  
     
     
         6 . The composition of  claim 1 , wherein the anti-CD28 antibody is 9.3 produced by a hybridoma assigned ATCC No. HB-10271.  
     
     
         7 . The composition of  claim 1 , wherein the anti-CD28 antibody is EX5.3D10 produced by a hybridoma assigned ATCC No. HB-11373.