Methods for induction of antigen-specific immune tolerance
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.
1. A method for inducing antigen-specific tolerance in a subject, comprising:
administering a tolerogenic composition to a subject in an amount effective to induce antigen-specific tolerance in the subject, wherein the tolerogenic composition comprises:
an immunogenic component comprising an antigen recognizable by the immune system of a subject, the immune system of the subject responding to the antigen with an unwanted immune response, wherein the immunogenic component is proinsulin or an immunogenic portion thereof; and
an erythrocyte-binding moiety coupled to the immunogenic component, wherein the erythrocyte-binding moiety non-covalently and specifically binds glycophorin A on erythrocytes and does not specifically bind to other blood components, wherein said erythrocyte-binding moiety comprises a peptide, an antibody, or antibody fragment; and
wherein upon administration of the tolerogenic composition, the tolerogenic composition binds specifically to erythrocytes via the cell binding component, and
wherein the cell binding component has a dissociation constant of between about 10 μM and 0.1 nM as determined by equilibrium binding measurements between the cell binding component and erythrocytes, and upon presentation of the immunogenic component to the immune system, antigen-specific tolerance is induced.
2. The method of claim 1 , wherein said administration is intravenous, wherein the immunogenic component is fused or chemically conjugated with the erythrocyte-binding moiety, wherein the erythrocyte-binding moiety comprises an antibody fragment, and wherein the erythrocyte-binding moiety is derived from a 10F7 clone.
3. The method of claim 2 , wherein the antibody fragment is affinity matured.
4. The method of claim 1 , wherein the erythrocyte-binding moiety is fused to the N- or C-terminus of the immunogenic component via a linker.
5. The method of claim 1 , wherein the erythrocyte-binding moiety is chemically conjugated to the immunogenic component.
6. The method of claim 1 , wherein the immunogenic component comprises a fragment of proinsulin.
7. The method of claim 1 , wherein the tolerogenic composition comprises a 10F7-derived antibody fragment- proinsulin fusion protein.
8. The method of claim 1 , wherein the erythrocyte-binding moiety comprises a peptide ligand, wherein the antigen comprises a fragment of proinsulin, wherein multiple copies of the erythrocyte-binding moiety are directly fused to the fragment of proinsulin at a plurality of sites on the fragment of proinsulin, and wherein the erythrocyte-binding moiety is capable of binding human glycophorin A.
9. A method for inducing antigen-specific tolerance in a subject, comprising:
administering a tolerogenic composition to a subject in an amount effective to induce antigen-specific tolerance in the subject, wherein the tolerogenic composition comprises:
an immunogenic component comprising an antigen recognizable by the immune system of a subject, the immune system of the subject responding to the antigen with an unwanted immune response, wherein the immunogenic component is a proinsulin or an immunogenic portion thereof comprising an epitope from proinsulin; and
an erythrocyte-binding moiety coupled to the immunogenic component, wherein the erythrocyte-binding moiety non-covalently and specifically binds glycophorin A on erythrocytes and does not specifically bind to other blood components, wherein said erythrocyte-binding moiety comprises an antibody fragment; and,
wherein upon administration of the tolerogenic composition, the tolerogenic composition binds specifically to erythrocytes via the erythrocyte-binding moiety and the immunogenic component is presented to the immune system, thereby inducing antigen-specific tolerance.
10. The method of claim 9 , wherein the immunogenic component comprises a fragment of proinsulin.
11. The method of claim 9 , wherein the immunogenic component comprises a mimotope of proinsulin.
12. The method of claim 9 , wherein said administration is intravenous.
13. The method of claim 9 , wherein the antibody fragment has a dissociation constant of between about 10 μM and 0.1 nM as determined by equilibrium binding measurements between the antibody fragment and erythrocytes, and wherein the antibody fragment is capable of binding human glycophorin A.
14. A method for inducing antigen-specific tolerance in a subject, comprising:
administering a tolerogenic composition to a subject, the tolerogenic composition comprising an immunogenic component coupled to an erythrocyte-binding moiety;
wherein the immunogenic component comprises an antigen recognizable by the immune system of a subject, the immune system of the subject responding to the antigen with an unwanted immune response,
wherein the immunogenic component is associated with development of Type I diabetes,
wherein the erythrocyte-binding moiety is configured to non-covalently, specifically bind glycophorin A on erythrocytes and not specifically bind to other blood components,
wherein said erythrocyte-binding moiety comprises an antibody fragment, or a single chain variable fragment (scFv) of antibody; and
wherein upon administration of the tolerogenic composition, the tolerogenic composition binds specifically to erythrocytes via the erythrocyte-binding moiety and the immunogenic component is presented to the immune system, thereby inducing antigen-specific tolerance.
15. The method of claim 14 , wherein said administration is intravenous, wherein the immunogenic component is chemically conjugated with the erythrocyte-binding moiety, wherein the erythrocyte-binding moiety has 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.
16. The method of claim 14 , wherein the erythrocyte-binding moiety is derived from a 10F7 clone.
17. The method of claim 14 , wherein the erythrocyte-binding moiety is an antibody, antibody fragment, or single chain variable fragment derived from an antibody, wherein the antibody, antibody fragment, or the single chain variable fragment is derived from a hybridoma that produces an antibody against a human erythrocyte.