Compositions and methods for inducing antigen-specific tolerance

Abstract

Erythrocyte-binding moieties coupled to tolerizing antigens are described. Provided for are peptidic ligands having sequences that specifically bind, or as antibodies or fragments thereof that provide specific binding, to erythrocytes. The erythrocyte-binding moieties may be prepared as molecular fusions with therapeutic agents, tolerizing antigens, or targeting peptides. Immunotolerance may be created by use of the fusions and choice of an antigen on a substance for which tolerance is desired.

What is claimed is: 
     
         1 . A composition for immunomodulation to achieve antigen-specific tolerance, the composition comprising:
 an antigen to which tolerance is desired;
 wherein the antigen is associated with rheumatoid arthritis; 
   an erythrocyte-binding moiety,
 wherein the erythrocyte-binding moiety has the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood, 
 wherein the erythrocyte-binding moiety comprises an antibody, antibody fragment, or a single chain variable fragment (scFv), 
 wherein the antigen to which tolerance is desired is recombinantly fused or chemically conjugated to the erythrocyte-binding moiety, and 
 wherein, upon administration to a human in which tolerance to the antigen is desired:
 the composition binds to CD45 negative cells, but not to CD45 positive cells, and the composition reduces, fails to induce, or prevents inflammatory responses in antigen-specific T cells as compared to when the human is exposed to the antigen alone. 
 
   
     
     
         2 . The composition of  claim 1 , wherein the erythrocyte-binding moiety is derived from a 10F7 clone, wherein the antibody fragment is affinity matured, and wherein the composition reduces the number of resident lymph node and spleen cells expressing interferon-gamma (IFNγ), as compared to the number of resident lymph node and spleen cells expressing IFNγ when the human is exposed to the antigen alone. 
     
     
         3 . The composition of  claim 1 , wherein the erythrocyte-binding moiety comprises an antibody fragment directed against human glycophorin A. 
     
     
         4 . The composition of  claim 3 , wherein the erythrocyte-binding moiety is derived from a 10F7 clone. 
     
     
         5 . The composition of  claim 3 , wherein the antibody fragment is affinity matured. 
     
     
         6 . The composition of  claim 1 , wherein the erythrocyte-binding moiety is fused via a linker, to the N- or C-terminus of the antigen. 
     
     
         7 . The composition of  claim 1 , wherein the administration of the composition ameliorates rheumatoid arthritis, and wherein the antigen is an immunogenic fragment of collagen II. 
     
     
         8 . A composition for immunomodulation to achieve antigen-specific tolerance, the composition comprising:
 an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety;   said antigen being recognizable by an immune system of a subject, the immune system of the subject being able to respond to the antigen with an unwanted immune response;   wherein said erythrocyte-binding moiety is capable of non-covalently and specifically binds an erythrocyte in situ in blood;   wherein the erythrocyte-binding moiety does not specifically bind to other blood components, wherein the other blood components comprises blood proteins, albumin, fibronectin, platelets, and white blood cells;   wherein said composition elicits a tolerogenic response upon administration to said subject; and   wherein said antigen is associated with multiple sclerosis.   
     
     
         9 . The composition of  claim 8 , wherein the antigen comprises a tolerogenic fragment of myelin oligodendrocyte glycoprotein. 
     
     
         10 . The composition of  claim 8 , wherein the erythrocyte-binding moiety is chemically conjugated to the antigen or wherein the erythrocyte-binding moiety is fused to the antigen via recombinant DNA technology. 
     
     
         11 . The composition of  claim 8 , wherein the erythrocyte-binding moiety comprises an antibody fragment, wherein the antibody fragment is derived from a 10F7 clone, and wherein the antibody fragment has undergone affinity maturation. 
     
     
         12 . A composition for immunomodulation to achieve antigen-specific tolerance, the composition comprising:
 an antigen fused or chemically conjugated with an erythrocyte-binding moiety;   wherein said is collagen II, a tolerogenic fragment of collagen II, or a combination thereof;   said antigen being recognizable by an immune system of a subject, the immune system of the subject being able to respond to or previously having responded to the antigen with an unwanted immune response,   said erythrocyte-binding moiety having the ability to non-covalently, specifically bind an erythrocyte surface in situ in blood and present said one or more antigens to the immune system of the subject,   wherein said erythrocyte-binding moiety comprises at least one peptide, an antibody, or an antibody fragment, and   wherein, upon administration to a human, the composition:   (i) binds to CD45 negative cells, but not to CD45 positive cells,   (ii) induces greater proliferation of antigen-specific CD8+ T cells, as compared to the proliferation of antigen-specific CD8+ T cells induced by the antigen alone, or   (iii) reduces the number of resident lymph node and spleen cells expressing interferon-gamma (IFNγ), as compared to the number of resident lymph node and spleen cells expressing IFNγ when the subject is exposed to the antigen alone.   
     
     
         13 . The composition of  claim 12 , wherein the erythrocyte-binding moiety has the ability to bind Band 3 (CD233), glycophorin-A, glycophorin B (CD235b), glycophorin C(CD235c), or glycophorin D (CD235d) with an affinity generating a dissociation constant of between about 10 μM and 0.1 nM as determined by equilibrium binding measurements between the erythrocyte-binding moiety and erythrocytes. 
     
     
         14 . The composition of  claim 12 , wherein the antigen is a tolerogenic portion of collagen II, wherein the erythrocyte-binding moiety does not specifically bind to other blood components, and wherein the other blood components comprises blood proteins, albumin, fibronectin, platelets, and white blood cells. 
     
     
         15 . The composition of  claim 12 , wherein the erythrocyte-binding moiety is derived from a 10F7 clone, and wherein the erythrocyte-binding moiety is affinity-matured. 
     
     
         16 . The composition of  claim 15 , wherein the erythrocyte-binding moiety is an antibody fragment. 
     
     
         17 . The composition of  claim 12 , wherein the administration of the composition ameliorates rheumatoid arthritis. 
     
     
         18 . The composition of  claim 12 , wherein the administration of the composition:
 (i) induces greater proliferation of antigen-specific CD8+ T cells, as compared to the proliferation of antigen-specific CD8+ T cells induced by the antigen alone and (ii) reduces the number of resident lymph node and spleen cells expressing interferon-gamma (IFNγ), as compared to the number of resident lymph node and spleen cells expressing IFNγ when the human is exposed to the antigen alone.   
     
     
         19 . The composition of  claim 12 , wherein the erythrocyte-binding moiety has the ability to bind human glycophorin-A. 
     
     
         20 . The composition of  claim 12 , wherein the erythrocyte-binding moiety has the ability to bind human glycophorin-A, wherein the erythrocyte-binding moiety is fused to the N- or C-terminus of the antigen, and wherein the antigen is a tolerogenic fragment of collagen II.