Method for polyclonal stimulation of t cells by mobile nanomatrices

Abstract

The present invention provides a method for polyclonal stimulation of T cells, the method comprising contacting a population of T cells with a nanomatrix, the nanomatrix comprising a) a matrix of mobile polymer chains, and b) attached to said matrix of mobile polymer chains one or more stimulatory agents which provide activation signals to the T cells; thereby activating and inducing the T cells to proliferate; wherein the nanomatrix is 1 to 500 nm in size. At least one first and one second stimulatory agents are attached to the same or to separate mobile matrices. If the stimulatory agents are attached to separate nanomatrices, fine-tuning of nanomatrices for the stimulation of the T cells is possible. Closed cell culture systems also benefit from this method.

1 . The use of a nanomatrix for in-vitro polyclonal stimulation of T cells, the nanomatrix comprising
 a) a matrix of mobile polymer chains, and   b) attached to said matrix of mobile polymer chains one or more stimulatory agents which provide activation signals to the T cells;   wherein the nanomatrix is 1 to 500 nm in size.   
     
     
         2 . An in vitro method for polyclonal stimulation of T cells, the method comprising contacting a population of T cells with a nanomatrix, the nanomatrix comprising
 a) a matrix of mobile polymer chains, and   b) attached to said matrix of mobile polymer chains one or more stimulatory agents which provide activation signals to the T cells; thereby activating and inducing the T cells to proliferate;   wherein the nanomatrix is 1 to 500 nm in size.   
     
     
         3 . The method according to  claim 2 , wherein at least one first and one second stimulatory agents are attached to the same matrix of mobile polymer chains. 
     
     
         4 . The method according to  claim 2 , wherein at least one first and one second stimulatory agents are attached to separate matrices of mobile polymer chains. 
     
     
         5 . The method according to  claim 4 , wherein the ratio of nanomatrices to cells is larger than 500:1 allowing fine-tuning of T cell stimulation. 
     
     
         6 . The method according to any one of  claims 2  to  5 , wherein one stimulatory agent is an anti-CD3 antibody or fragment thereof. 
     
     
         7 . The method according to any one of  claims 1  to  6 , wherein the second stimulatory agent is an anti-CD28 antibody. 
     
     
         8 . The method according to any one of  claims 2  to  7 , wherein the matrix of mobile polymer chains consists of a polymer of dextran. 
     
     
         9 . The method according to any one of  claims 2  to  8 , wherein the nanomatrix carries magnetic, paramagnetic or superparamagnetic nano-crystals, embedded into the matrix of mobile polymer chains. 
     
     
         10 . The method according to any one of  claims 2  to  9 , wherein the stimulatory agent is attached at high density with more than 25 μg per mg nanomatrix. 
     
     
         11 . The method according to any one of  claims 2  to  10 , wherein the stimulated T cells are Treg cells. 
     
     
         12 . A method for polyclonal stimulation of T cells in a closed cell culture system, the method comprising contacting a cell culture comprising a population of T cells within said closed cell culture system with a dosage of nanomatrix, the nanomatrix comprising
 a) a matrix of mobile polymer chains; and   b) attached to said matrix of mobile polymer chains one or more stimulatory agents which provide activation signals to the T cells;   wherein the nanomatrix is 1 to 500 nm in size, and   wherein said dosage of nanomatrix is applied sterile to said closed cell culture system, and   wherein said dosage depends on the volume of the cell culture in said closed cell culture system.   
     
     
         13 . A method according to  claim 12 , wherein said volume of the cell culture can be determined by a balance or a camera system without affecting the sterility barrier. 
     
     
         14 . A method according to  claims 12  and  13 , wherein determining and applying said dosage are performed automatically. 
     
     
         15 . A pharmaceutical composition comprising a population of stimulated T cells produced according to the method of  claims 12 - 14 .