Erythrocyte-binding therapeutics
Peptides that specifically bind erythrocytes are described. These are provided as peptidic ligands having sequences that specifically bind, or as antibodies or fragments thereof that provide specific binding, to erythrocytes. The peptides 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. Fusions with targeting peptides direct the fusions to the target, for instance a tumor, where the erythrocyte-binding ligands reduce or entirely eliminate blood flow to the tumor by recruiting erythrocytes to the target.
1. A method for inducing antigen-specific tolerance to an antigen in a subject, the method comprising:
administering a composition to a subject in an amount effective to induce antigen-specific tolerance, the composition comprising:
i) 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 fragment directed against glycophorin A, and
ii) an antigen to which tolerance is desired, wherein the antigen is selected from a portion of myelin basic protein, a portion of myelin oligodendrocyte glycoprotein, a portion of proteolipid protein, myelin basic protein, myelin oligodendrocyte glycoprotein, proteolipid protein, and combinations thereof;
wherein the antigen to which tolerance is desired is recombinantly fused 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.
2. The method of claim 1 wherein the administration is intravenous, 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.
3. The method of claim 1 wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N-terminus of the antigen.
4. The method of claim 3 wherein the erythrocyte-binding moiety is derived from 10F7.
5. The method of claim 1 wherein the erythrocyte-binding moiety is affinity matured.
6. The method of claim 1 wherein the erythrocyte-binding moiety is derived from 10F7 and the antigen to which tolerance is desired is a portion of myelin basic protein.
7. The method of claim 6 wherein the inducing antigen-specific tolerance ameliorates multiple sclerosis.
8. A method for inducing antigen-specific tolerance to an antigen in a subject, the method comprising:
administering a composition to a subject in an amount effective to induce antigen-specific tolerance, the composition comprising an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety,
wherein the erythrocyte-binding moiety comprises an antibody fragment or a peptide ligand that specifically binds to human erythrocytes;
wherein the antigen is an antigen to which a subject develops an unwanted immune response, wherein the antigen is associated with multiple sclerosis; 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.
9. The method of claim 8 , wherein the peptide ligand comprises SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17.
10. The method of claim 9 , wherein the antigen comprises a portion of myelin oligodendrocyte glycoprotein (MOG).
11. The method of claim 10 , wherein the erythrocyte-binding moiety comprises an antibody fragment, wherein the antibody fragment comprises an affinity matured antibody fragment.
12. The method of claim 11 wherein the erythrocyte-binding moiety is derived from 10F7.
13. The method of claim 8 wherein the inducing antigen-specific tolerance ameliorates multiple sclerosis.
14. A method for inducing antigen-specific tolerance to an antigen in a subject, the method comprising:
administering a composition to a subject in an amount effective to induce antigen-specific tolerance, the composition comprising an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety;
wherein the erythrocyte-binding moiety comprises an antibody, antibody fragment, or a single chain variable fragment (scFv) that binds to human glycophorin A; and
wherein the antigen is a self-antigen to which a subject develops an unwanted immune response, and
wherein the self-antigen is associated with multiple sclerosis.
15. The method of claim 14 , wherein the self-antigen is selected from a portion of myelin oligodendrocyte glycoprotein, a portion of myelin basic protein, a portion of proteolipid protein, myelin basic protein, myelin oligodendrocyte glycoprotein, proteolipid protein, and combinations thereof.
16. The method of claim 14 , wherein the erythrocyte-binding moiety is fused to the antigen via recombinant DNA technology.
17. The method of claim 14 , wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N- or C-terminus of the antigen.
18. The method of claim 14 , wherein the erythrocyte-binding moiety comprises an antibody fragment, and wherein the erythrocyte-binding moiety is derived from a 10F7 clone.
19. The method of claim 14 , wherein the erythrocyte-binding moiety is an antibody fragment, wherein the antibody fragment is affinity matured, and wherein the antigen to which tolerance is desired comprises a portion of myelin oligodendrocyte glycoprotein (MOG).
20. The method of claim 14 , wherein the erythrocyte-binding moiety is an antibody fragment, wherein the antibody fragment is affinity matured, and wherein the antigen to which tolerance is desired comprises a portion of myelin basic protein.