Tolerogenic dendritic cells, methods of producing the same, and uses thereof

Abstract

Described herein are methods for the preparation of stable semi-mature tolerogenic dendritic cells and compositions comprising such stable semi-mature tolerogenic dendritic cells. The stable semi-mature tolerogenic dendritic cells described herein and compositions thereof can be used for the establishment of immune tolerance when treating an autoimmune disease, graft rejection and/or graft-versus-host disease.

1 - 74 . (canceled) 
     
     
         75 . A population of stable semi-mature tolerogenic dendritic cells produced by a method comprising the following steps:
 a. culturing monocytes isolated from a subject's blood in culture medium comprising GM-CSF and IL-4;   b. after a first period of time in culture, culturing the cells from step (a) in culture medium comprising Dexamethasone, GM-CSF and IL-4;   c. after a second period of time in culture, culturing the cells from step (b) in culture medium comprising Dexamethasone, vitamin D2, GM-CSF and IL-4 for a third period of time to generate tolerogenic dendritic cells; and   d. after third period of time in culture, culturing the tolerogenic dendritic cells in culture medium comprising MPLA, GM-CSF and IL-4, or MPLA, GM-CSF, IL-4 and an antigen(s) associated with an autoimmune disease, graft rejection or graft-versus-host disease;   wherein the cells from step (a) are cultured in culture medium comprising Dexamethasone, GM-CSF and IL-4 on the 3 rd  day in culture, and   wherein the cells from step (b) are cultured in culture medium comprising Dexamethasone, vitamin D2, GM-CSF and IL-4 on the 6 th  day of culture, and   wherein the tolerogenic dendritic cells are cultured in culture medium comprising MPLA, GM-CSF and IL-4, or GM-CSF, IL-4, MPLA and the antigen(s) on the 7 th  day of culture,   and wherein the tolerogenic dendritic cells induce a higher number of CD4 + CD25 + FoxP3 +  regulatory T cells than dendritic cells obtained by culturing monocytes in culture medium comprising GM-CSF and IL-4 without Dexamethasone and vitamin D2.   
     
     
         76 . The population of stable semi-mature tolerogenic dendritic cells of  claim 75 , wherein the antigen(s) is associated with an autoimmune disease, and further wherein the tolerogenic dendritic cells induce a higher number of CD4 + CD25 + FoxP3 +  regulatory T cells than dendritic cells obtained by culturing monocytes in culture medium comprising GM-CSF and IL-4 without Dexamethasone and vitamin D2. 
     
     
         77 . The population of stable semi-mature tolerogenic dendritic cells of  claim 76 , wherein the autoimmune disease is type 1 diabetes, and further wherein the tolerogenic dendritic cells induce a higher number of CD4 + CD25 + FoxP3 +  regulatory T cells than dendritic cells obtained by culturing monocytes in culture medium comprising GM-CSF and IL-4 without Dexamethasone and vitamin D2. 
     
     
         78 . The population of stable semi-mature tolerogenic dendritic cells of  claim 75 , wherein the antigen(s) is associated with graft rejection or graft-versus-host disease, and further wherein the tolerogenic dendritic cells induce a higher number of CD4 + CD25 + FoxP3 +  regulatory T cells than dendritic cells obtained by culturing monocytes in culture medium comprising GM-CSF and IL-4 without Dexamethasone and vitamin D2.