Uses of bispecific antibody coated dendritic cells pulsed with antigens and gm-csf in immune regulation

Abstract

GM-CSF administered before immunization exerted a sustained suppressive effect against the induction of myasthenia gravis (MG). This suppression was associated with lowered serum autoantibody levels, reduced T cell proliferative responses to AChR, and an expansion in the population of FoxP3+ regulatory T cells. Manipulating DCs to expand regulatory T cells is useful for the control of autoimmune diseases such as myasthenia gravis MG.

1 . A composition comprising a semi-mature dendritic cell coated with a bispecific antibody, wherein the bispecific antibody is capable of binding to a specific antigen of interest and a T-cell receptor. 
   
   
       2 . The composition of  claim 1 , wherein the bispecific antibody is an anti-CTLA-4 bispecific antibody. 
   
   
       3 . The composition of  claim 1 , wherein the dendritic cell is treated with GM-CSF. 
   
   
       4 . The composition of  claim 1 , wherein the dendritic cell is pulsed with an antigen of interest. 
   
   
       5 . A bispecific anti-CTLA-4 antibody coated to a semi-mature dendritic cell, wherein the antibody is also capable of binding to a specific antigen. 
   
   
       6 . A method to induce antigen specific T-cell tolerance for treating an autoimmune disease, the method comprising:
 (a) contacting a dendritic cell with GM-CSF to obtain a semi-mature dendritic cell;   (b) pulsing the semi-mature dendritic cell with an antigen of interest;   (c) coating the pulsed semi-mature dendritic cell with a bispecific antibody capable of binding a T-cell receptor on an activated T-cell; and   (d) modulating the production of a regulatory T-cell to induce T-cell tolerance.   
   
   
       7 . The method of  claim 6 , wherein the bispecific antibody is capable of binding the T-cell receptor CTLA-4. 
   
   
       8 . The method of  claim 6 , wherein the bispecific antibody is anti-CTLA-4 antibody. 
   
   
       9 . The method of the  claim 6 , wherein the antigen of interest is an allo or a self antigen. 
   
   
       10 . The method of  claim 6 , wherein the semi-mature dendritic cell induces a regulatory T-cell production. 
   
   
       11 . The method of  claim 6 , further comprising infusing the regulatory T cell into a host to suppress allo graft rejection or to treat an autoimmune disease. 
   
   
       12 . The method of  claim 6 , wherein the GM-CSF and the bispecific antibody are introduced in vivo to modulate the production of the regulatory T-cell. 
   
   
       13 . The method of  claim 6 , wherein the disease is selected from the group consisting of rheumatoid arthritis, systemic sclerosis, lupus, Hashimoto's thyroiditis, Type-1 diabetes, Myasthenia gravis, atopic dermatitis, and multiple sclerosis. 
   
   
       14 . A method to treat a T and B cell mediated autoimmune disease and reduce the risk of a graft rejection by the host in a mammal, the method comprising:
 (a) presenting an antigen responsible for the autoimmune disease or the graft rejection by the host to a dendritic cell;   (b) culturing bispecific antibody coated dendritic cells with a population of T-cells to induce antigen specific T-cell tolerance, wherein the bispecific antibody is capable of binding to a dendritic cell associated with the antigen and also with a T-cell through a T-cell receptor; and   (c) administering the population of T-cells to the host and the bispecific antibody coated antigen bound dendritic cells to treat the autoimmune disease and to reduce the graft rejection.   
   
   
       15 . The method of  claim 14 , wherein the T-cell receptor is CTLA-4 
   
   
       16 . A method to affect maturation of dendritic cells, wherein the cells are held in semi-mature status that can induce regulatory T cells, the method comprising
 (a) pulsing dendritic cells;   (b) contacting the dendritic cells with GM-CSF; and   (c) coating the pulsed dendritic cells with an appropriate bi-specific antibody, wherein one arm provides target or antigen specificity and the other arm provides CTLA-4 ligation.   
   
   
       17 . The method of  claim 16 , wherein the regulatory T cells (Tregs) are either generic or antigen specific. 
   
   
       18 . The method of  claim 16 , wherein regulatory T cells produce TGF-beta and/or IL-10 through which Tregs can suppress immune responses. 
   
   
       19 . A method to treat a T- and B-cell mediated autoimmune disease and prevent allograft rejection, the method comprising:
 (a) treating dendritic cells with GM-CSF to maintain the dendritic cells in a semi-mature state in the presence of an antigen of interest, wherein the antigen is an alloantigen or an autoantigen;   (b) inducing the semi-mature dendritic cells in vitro or in vivo;   (c) using semi-mature dendritic cells either in vivo or in vitro as antigen presenting cells to induce regulatory T-cells capable of producing IL-10 and/or TGF-beta to suppress effector T cell functions; and   (d) treating the autoimmune disease with the regulatory T cells, wherein the regulatory T cells are induced in vivo with GM-CSF followed by a treatment with a bi-specific antibody to suppress T-cell function.   
   
   
       20 . The method of  claim 19 , wherein the disease is selected from the group consisting of rheumatoid arthritis, systemic sclerosis, lupus, Hashimoto's thyroiditis, Type-1 diabetes, Myasthenia gravis, atopic dermatitis, and multiple sclerosis. 
   
   
       21 . The method of  claim 19 , wherein the regulatory T cells are administered in vivo. 
   
   
       22 . A method of treating an autoimmune disease comprising administering to a patient in need of treatment, a therapeutically effective amount of a composition comprising antigen-activated semi-mature dendritic cells, wherein the dendritic cells are produced by a method comprising culturing a tissue source in a culture medium, comprising GM-CSF and at least one other factor selected from the group consisting of IL-10, IL-4 or TGFβ, which inhibits proliferation or maturation of non-dendritic cell precursors, thereby increasing the proportion of dendritic cell precursors in the culture;
 wherein the dendritic cells are pulsed with a self-protein or alloantigen, and wherein the dendritic cells process the self-protein to produce a modified self-protein antigen or alloantigen, which is expressed by the dendritic cells;   coating the semi-mature antigen bearing dendritic cells with a bi-specific antibody that is capable of binding to a dendritic cell associated with an antigen and also with a T-cell through a T-cell receptors; and   administering the semi-mature dendritic cells into the host.