Methods and compositions for immunomodulation

Abstract

Provided are cells containing exogenous antigen and uses thereof.

What is claimed is: 
     
         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.