Methods and compositions for sustained immunotherapy

Abstract

This disclosure provides methods of making functionalized PEG iron oxide nanoparticles.

What is claimed: 
     
         1 . A complex comprising a nanoparticle and disease-relevant antigen-MHCII complexes for use in expanding and/or developing populations of Tr1 cells and/or B-regulatory cells in subject, wherein the nanoparticle has a diameter selected from the group of: from about 1 nm to about 100 nm in diameter; from about 1 nm to about 50 nm in diameter or from about 1 nm to about 20 nm or from about 5 nm to about 100 nm in diameter and wherein the ratio of the number of antigen-MHCII complexes to nanoparticle is from about 10:1 to about 1000:1. 
     
     
         2 . The complex of  claim 1 , wherein the complex has an antigen-MHCII density from about 0.05 pMHC/100 nm 2  of the surface area of the nanoparticle to about 25 pMHC/100 nm 2  of the surface area of the nanoparticle. 
     
     
         3 . The complex of  claim 1  or  2 , wherein the antigen is derived from an autoantigen involved in an autoimmune response or mimic thereof, and optionally wherein the autoantigen is an epitope from an antigen expressed by pancreatic beta cells, IGRP, Insulin, GAD, IA-2 or myelin oligodendrocyte protein (MOG). 
     
     
         4 . The complex  claim 1 , wherein the nanoparticle is non-liposomal and/or has a solid core, preferably a gold or iron oxide core. 
     
     
         5 . The complex of  claim 1 , wherein the antigen-MHCII is covalently or non-covalently linked to the nanoparticle. 
     
     
         6 . The complex of  claim 1 , wherein the antigen-MHCII is covalently linked to the nanoparticle through a linker less than 5 kD in size. 
     
     
         7 . The complex of  claim 1 , wherein the nanoparticle is bioabsorbable and/or biodegradable. 
     
     
         8 . The complex of  claim 1 , wherein the nanoparticle complex comprises antigen-MHCII complexes to nanoparticle ratio of from about 10:1 to about 100:1. 
     
     
         9 . The complex of  claim 5 , wherein the linker comprises polyethylene glycol. 
     
     
         10 . The complex of  claim 1 , wherein the antigen-MHCII complexes are identical or different. 
     
     
         11 . The complex of  claim 9  or  10 , wherein the linkers are identical or different. 
     
     
         12 . A composition comprising a therapeutically effect amount of the complex of  claim 1  and a carrier. 
     
     
         13 . The composition of  claim 12 , wherein the carrier is a pharmaceutically acceptable carrier. 
     
     
         14 . A method for making, preparing or obtaining the complex of  claim 1 , comprising coating or complexing antigen-MHCII complexes onto a nanoparticle. 
     
     
         15 . A method for promoting the formation, expansion and recruitment of Tr1 cells and/or B-regulatory cells in an antigen-specific manner in a subject in need thereof, comprising administering to the subject an effective amount of the complex of  claim 1 . 
     
     
         16 . A method for treating or preventing a viral infection or an autoimmune disorder in a subject in need thereof comprising administering to the subject an effective amount of the complex of  claim 1 , with the proviso that the antigen is a autoimmune-relevant autoantigen. 
     
     
         17 . The method of any one of  claims 14 - 16 , wherein the subject is a mammal. 
     
     
         18 . The method of  claim 15 , wherein the autoimmune disorder is selected from the group of diabetes, pre-diabetes, multiple sclerosis or a multiple sclerosis-related disorder, with the proviso that the antigen is relevant to the autoimmune disorder being treated. 
     
     
         19 . A kit comprising the complex of  claim 1 , and instructions for use. 
     
     
         20 . A method for making functionalized PEG iron oxide nanoparticles comprising thermally decomposing iron acetyl acetonate in the presence of functionalized PEG molecules and benzyl ether, wherein the thermal decomposition occurs at a temperature from about 80 to about 300° C. 
     
     
         21 . The method of  claim 20 , wherein the iron oxide nanoparticle is water-soluble. 
     
     
         22 . The method of  claim 20 , wherein the thermal decomposition comprises a single-step reaction. 
     
     
         23 . The method of  claim 20 , wherein the thermal decomposition occurs in the presence of functionalized PEG molecules. 
     
     
         24 . The method of  claim 23 , wherein the thermal decomposition is carried out in the presence of benzyl ether. 
     
     
         25 . The method of  claim 20 , wherein the temperature for the thermal decomposition is about 80 to about 200° C., or about 80 to about 150° C., or about 100 to about 250° C., or about 100 to about 200° C., or about 150 to about 250° C., or about 150 to about 250° C. 
     
     
         26 . The method of  claim 20 , wherein the thermal decomposition is carried out for about 1 to about 2 hours. 
     
     
         27 . The method of  claim 20 , wherein the nanoparticles are stable at about 4° C. in PBS without any detectable degradation or aggregation. 
     
     
         28 . The method of  claim 27 , wherein the nanoparticles are stable for at least 6 months. 
     
     
         29 . The method of  claim 20 , wherein the method further comprises purifying the nanoparticles with a magnetic column. 
     
     
         30 . A method for making nanoparticle complexes comprising contacting pMHC with iron oxide nanoparticles as obtained from  claim 20 , thereby providing nanoparticle complexes. 
     
     
         31 . The method of  claim 30 , further comprising purifying the nanoparticles with a magnetic column. 
     
     
         32 . The method of  claim 20 , wherein the functionalized PEG molecules are maleimide functionalized. 
     
     
         33 . The method of  claim 20 , wherein the functionalized PEG molecules are less than about 5 kilodaltons. 
     
     
         34 . The method of  claim 32 , wherein the maleimide functionalized PEG molecules are methoxy-PEG molecules.