Therapeutic vaccine for treatment of diabetes type 1 in children, application of the cell sorter and the method of multiplying treg cells to produce therapeutic vaccine for treatment of diabetes type 1

Abstract

The gist of the invention consists in the therapeutic vaccine for treatment of diabetes type 1 in children, which contains Treg cells CD3(+)CD4(+)CD25(high)CD127(−). Claimed too is the cell sorter used to produce the vaccine and the method of multiplying Treg cells in vitro.

1 . Therapeutic vaccine for treatment of diabetes type 1 in children characterised in that it contains:
 Treg cells CD3(+)CD4(+)CD25(high)CD127(−)   
     
     
         2 . Application of the cell sorter to produce the vaccine defined in  claim 1 , characterised in that the cells are sorted to isolate Treg cells using the algorithm sorting the following phenotype:
 CD3(+)CD4(+)CD25(high)CD127(−)doublet(−)lineage(−)dead(−).   
     
     
         3 . The method of multiplying Treg cells in vitro to produce the vaccine defined in  claim 1 , characterised in that:
 lymphocytes T CD4+ are isolated by the immunomagnetic method and marked with monoclonal antibodies,   the sorter-isolated Treg cells are multiplied in the CellGro CellGro or X-VIVO medium supplemented with autological inactivate serum and interleukin-2,   the culture is added artificial antigen-presenting cells in the 1:1 proportion.   
     
     
         4 . The method according to  claim 3 , characterised in that selected for the marking are the following monoclonal antibodies: anti-CD3, anti-CD4, anti-CD8, anti-CD19, anti-CD14, anti-CD16, anti-CD25, and anti-CD127. 
     
     
         5 . The method according to  claim 3 , characterised in that the monoclonal antibodies CD3, CD4, CD8, CD19, CD14, CD16, CD25, and CD127 recognise the antigens, and are conjugated with fluorescent dyes. 
     
     
         6 . The method according to  claim 3 , characterised in that the antigen-presenting artificial cells are magnetic beads coated with anti-CD3 and anti-CD28 antibodies.