Compositions for inducing antigen-specific 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. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:
an antigenic component comprising asparaginase coupled to a cell binding component;
the antigenic component comprising a polypeptide capable of eliciting an immune response from the immune system of the subject;
the cell binding component comprising an antibody fragment, the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,
the cell binding component being configured to bind glycophorin A on the surface of human erythrocytes; and
wherein the cell binding component is an antibody fragment derived from a hybridoma,
wherein the antibody fragment is affinity matured,
wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes,
wherein the cell binding component exhibits an erythrocyte equilibrium 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
wherein, upon administration of the engineered composition, the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.
2. The composition of claim 1 , wherein the cell binding component is derived from a 10F7 clone, wherein the antigenic portion is coupled to the cell binding component as a molecular fusion, and wherein the composition reduces, fails to induce or prevents inflammatory responses in antigen-specific T cells when the subject is exposed to the antigenic component alone.
3. The composition of claim 1 , wherein the cell binding component is derived from a 10F7 clone.
4. The composition of claim 1 , wherein the cell binding component is fused to the N- or C-terminus of the antigenic component via a linker.
5. The composition of claim 1 , wherein the cell binding component is chemically conjugated to the antigenic component.
6. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:
an antigenic component coupled to a cell binding component;
the antigenic component comprising asparaginase or a polypeptide derived from asparaginase and capable of eliciting an immune response from the immune system of a subject;
the antigenic component being fused with the cell binding component;
the cell binding component comprising a polypeptide,
the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,
the cell binding component being configured to bind to glycophorin A on the surface of erythrocytes with an equilibrium dissociation constant of between about 10 μM and 0.1 nM, as determined by equilibrium binding measurements between the cell binding component and erythrocytes;
wherein the cell binding component is an antibody fragment derived from a hybridoma,
wherein the antibody fragment is affinity matured,
wherein the engineered composition is suitable for intravenous administration to the subject; and
wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes, and the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.
7. The composition of claim 6 , wherein antigenic component comprises a fragment of asparaginase.
8. The composition of claim 6 , wherein the antigenic portion is a mimotope of asparaginase.
9. The composition of claim 6 , wherein cell binding component is fused to the N-terminus of the antigenic component.
10. The composition of claim 6 , wherein cell binding component is fused to the antigenic component via a linker.
11. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:
an antigenic component coupled to a cell binding component;
the antigenic component comprising a polypeptide that is capable of eliciting an immune response from the immune system of the subject, said antigenic component being chemically conjugated with the cell binding component;
the cell binding component comprising a polypeptide, the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,
the cell binding component being configured to bind glycophorin A on the surface of erythrocytes,
wherein the cell binding component comprises an antibody fragment; and
wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes, the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.
12. The composition of claim 11 , wherein the antigenic component comprises asparaginase.
13. The composition of claim 11 , wherein the cell binding component is derived from a 10F7 clone.
14. An engineered composition for inducing antigen-specific tolerance in a subject, the engineered composition comprising:
an antigenic component coupled to a cell binding component;
the antigenic component comprising asparaginase or a polypeptide derived from asparaginase and capable of eliciting an immune response from the immune system of a subject;
the antigenic component being fused with the cell binding component;
the cell binding component comprising a polypeptide,
the cell binding component being configured to specifically bind erythrocytes and not specifically bind to other blood components,
the cell binding component comprises a sequence chosen from the group consisting of SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:1, and conservative substitutions thereof,
the cell binding component being configured to bind to glycophorin A on the surface of erythrocytes with an equilibrium dissociation constant of between about 10 μM and 0.1 nM, as determined by equilibrium binding measurements between the cell binding component and erythrocytes;
wherein the engineered composition is suitable for intravenous administration to the subject; and
wherein upon administration of the engineered composition, the cell binding component binds to erythrocytes, and the antigenic component is presented to the immune system and tolerance specific to the antigenic component is induced.
15. The composition of claim 14 , wherein multiple copies of the cell binding component are directly fused to the antigen at a plurality of sites on the antigenic component.
16. The composition of claim 14 , wherein antigenic component comprises a fragment of asparaginase.
17. The composition of claim 14 , wherein the antigenic portion is a mimotope of asparaginase.
18. The composition of claim 14 , wherein cell binding component is fused to the N-terminus of the antigenic component.
19. The composition of claim 14 , wherein cell binding component is fused to the antigenic component via a linker.