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 composition for use in inducing tolerance comprising:
an antigen to which tolerance is desired;
wherein the antigen is selected from the group consisting of insulinoma-associated protein 2 (IA-2) and an immunogenic portion of said IA-2;
an erythrocyte-binding moiety,
wherein the erythrocyte-binding moiety comprises an antibody fragment that has the ability to non-covalently, specifically bind an exterior erythrocyte surface in situ in blood,
wherein the erythrocyte-binding moiety is affinity matured,
wherein the antigen to which tolerance is desired is recombinantly fused to the erythrocyte-binding moiety,
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, 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.
2. The composition of claim 1 wherein the erythrocyte-binding moiety is fused, optionally via a linker, to the N-terminus of the antigen.
3. The composition of claim 2 wherein the erythrocyte-binding moiety is derived from 10F7.
4. The composition of claim 1 wherein the erythrocyte-binding moiety is derived from 10F7 and the antigen to which tolerance is desired is an immunogenic portion of said IA-2.
5. The composition of claim 4 wherein the administration of the composition ameliorates Type I Diabetes.
6. A composition comprising an antigen recombinantly fused or chemically conjugated with an erythrocyte-binding moiety;
wherein the erythrocyte-binding moiety comprises an antibody fragment that specifically binds to human erythrocytes; and
wherein the antigen is a self-antigen to which a subject develops an unwanted immune response,
wherein the self-antigen is associated with type 1 diabetes, 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.
7. The composition of claim 6 , wherein the self-antigen is selected from the group consisting of glutamic acid decarboxylase 65 (GAD-65), GAD-67, insulinoma-associated protein 2 (IA-2), insulinoma-associated protein 2β (IA-2b), islet-specific glucose-6-phosphatase catalytic subunit-related protein, a portion of any one of said GAD-65, said GAD-67, said IA-2, said IA-2b, and said islet-specific glucose-6-phosphatase catalytic subunit-related protein, and mimotopes thereof.
8. The composition of claim 6 , wherein the erythrocyte-binding moiety is fused to the antigen via recombinant DNA technology.
9. The composition of claim 6 , wherein the erythrocyte-binding moiety comprises an antibody fragment, and wherein the erythrocyte-binding moiety is derived from a 10F7 clone.
10. A composition for use in inducing tolerance comprising:
an antigen to which tolerance is desired;
wherein the antigen is a self-antigen to which a subject develops an unwanted immune response,
and wherein the self-antigen is associated with type 1 diabetes;
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 or antibody fragment directed against glycophorin A,
wherein, upon administration to a human in which tolerance to the antigen is desired, 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, 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.
11. The composition of claim 10 wherein the self-antigen is selected from the group consisting of glutamic acid decarboxylase 65 (GAD-65), GAD-67, insulinoma-associated protein 2 (IA-2), insulinoma-associated protein 2β (IA-2b), islet-specific glucose-6-phosphatase catalytic subunit-related protein, portions thereof, and mimotopes thereof.