METHODS AND COMPOSITIONS FOR IMMUNOMODULATION
Provided are cells containing exogenous antigen and uses thereof.
1 . A method of making an enucleated erythroid cell comprising an exogenous antigen, the method comprising:
subjecting an enucleated erythroid cell to a controlled cell injury in the presence of the exogenous antigen;
thereby making the enucleated erythroid cell comprising an exogenous antigen.
2 . The method of claim 1 , wherein the controlled cell injury caused a perturbation in the cell membrane of the enucleated erythroid cell.
3 . The method of claim 1 , wherein the controlled cell injury comprises electroporation, sonoporation, liposomal transfection, or salt-based transfection.
4 . The method of claim 1 , wherein the controlled cell injury comprises cell deformation.
5 . The method of claim 1 , wherein the controlled cell injury comprises cell squeezing.
6 . The method of claim 1 , wherein the controlled cell injury comprises passing the enucleated erythroid cell through a micro-channel.
7 . The method of claim 6 , which comprising moving the cell through the micro-channel by application of pressure.
8 . The method of claim 1 , wherein the controlled cell injury results in the cell membrane becoming porous.
9 . The method of claim 1 , which further comprising isolating an enucleated erythroid cell from a subject before subjecting the enucleated erythroid cell to a controlled cell injury.
10 . The method of claim 1 , further comprising autologous administration of the enucleated erythroid cell comprising the exogenous antigen to the subject.
11 . The method of claim 1 , wherein the exogenous antigen is intracellular.
12 . The method of claim 1 , wherein the exogenous antigen is present in the cytoplasm of the enucleated erythroid cell.
13 . The method of claim 1 , wherein the enucleated erythroid cell is not a hypotonically dialysed cell.
14 . The method of claim 1 , wherein a pharmaceutical composition comprising a plurality of the enucleated erythroid cell comprising an exogenous antigen is capable of inducing immune tolerance in a human subject suffering from or at risk of developing an autoimmune disease, disorder or condition.
15 . The method of claim 1 , wherein the enucleated erythroid cell comprises at least 1,000 copies of the exogenous antigen.
16 . The method of claim 1 , wherein the exogenous antigen is selected from insulin, proinsulin, preproinsulin, GAD65, IA-2, myelin oligodendrocyte glycoprotein, myelin basic protein, and proteolipid protein, or an antigenic fragment thereof.
17 . The method of claim 1 , wherein the enucleated erythroid cell is a mature erythrocyte.
18 . The method of claim 1 , wherein the enucleated erythroid cell is a reticulocyte.
19 . The method of claim 1 , which comprises a population of erythroid cells that is greater than 80% enucleated.
20 . The method of claim 1 , wherein the enucleated erythroid cells have a viability that is greater than 70%.
21 . The method of claim 1 , wherein at least 80% of cells in the composition comprise the exogenous antigen and are enucleated.