Process for t cell expansion

Abstract

An in vitro expansion process for rapid expansion of antigen specific T cells, such as allogeneic antigen specific cells comprising the steps culturing in a gas permeable vessel a population of PBMCs (such as allogeneic PBMCs) in the presence of antigen, for example a peptide or peptide mix relevant to a target antigen(s), in the presence of an exogenous cytokine characterized in that the expansion to provide the desired population of T cells is 14 days or less, for example 9, 10, 11 or 12 days, such as 10 days. The disclosure also extends to T cell populations generated by and obtained from the method and the use of same in therapy.

1 . An in vitro expansion process for rapid expansion of antigen specific T cells (such as allogeneic antigen specific T cells) comprising the steps culturing in a gas permeable vessel a population of PBMCs (such as allogeneic PBMCs) in the presence of a peptide or peptide mix relevant to a target antigen(s), in the presence of an exogenous cytokine characterised in that the cytokine is other than exogenous IL-2. 
     
     
         2 . An in vitro expansion process for rapid expansion of antigen specific T cells, such as allogeneic antigen specific T cells comprising the steps culturing in a gas permeable vessel a population of PBMCs (such as allogeneic PBMCs) in the presence of antigen, for example a peptide or peptide mix relevant to a target antigen(s), in the presence of an exogenous cytokine characterised in that the expansion to provide the desired population of T cells is 14 days or less, for example 9, 10, 11 or 12 days, such as 10 days. 
     
     
         3 . An in vitro expansion process according to  claim 1 , wherein the exogenous cytokine is selected from the group comprising IL-4, IL-7, IL-15 or a combination thereof. 
     
     
         4 . An in vitro expansion process according to  claim 2 , wherein the exogenous cytokine is selected from the group comprising IL-4, IL-7, IL-15 or a combination thereof. 
     
     
         5 . An in vitro expansion process according to  claim 3  or  4 , wherein the exogenous cytokine is a combination of IL-4 and IL-7. 
     
     
         6 . An in vitro process according to any one of  claims 1  to  5 , wherein the culture is performed in the presence of a T cell expansion medium. 
     
     
         7 . An in vitro process according to any one of  claims 1  to  6 , wherein the peptide mix comprises between 2 and 500 peptides, for example 2 to 400 peptides, 2 to 300 or 2 to 200 peptides. 
     
     
         8 . An in vitro process according to  claim 7 , wherein the peptides overlap, for example by 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more amino acids. 
     
     
         9 . An in vitro process according to any one of  claims 1 , to  8 , wherein the peptides are on average about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids in length, such as 15 amino acids in length. 
     
     
         10 . An in vitro process according to any one of  claims 1  and  3  to  9 , wherein the expansion is performed for 14 days or less. 
     
     
         11 . An in vitro process according to any one of  claims 1  to  10  wherein the expansion is performed for 13, 12, 11, 10 days or less. 
     
     
         12 . An in vitro process according to any one of  claims 1  to  11 , wherein the initial PBMC sample derived from a donor is a mobilised sample or mobilised apheresis sample. 
     
     
         13 . An in vitro process according to any one of  claims 1  to  12 , wherein the gas permeable vessel is a GRex system or a derivative or an equivalent system. 
     
     
         14 . An in vitro process according to  claim 13  wherein the GRex system has been adapted to provide a closed system, suitable for aseptic manufacture, for example as shown in  FIG. 3 . 
     
     
         15 . An in vitro process according to any one of  claims 1  to  14 , wherein the process is performed aseptically or in a clean room. 
     
     
         16 . An in vitro process according to any one of  claims 1  to  15 , wherein media and nutrients are not added or changed after initiation of the expansion process. 
     
     
         17 . A process according as defined in any one of  claims 1  to  16 , which comprises the further step of preparing a pharmaceutically acceptable composition comprising the further step of adding a diluent, stabilizer, preservative and/or other pharmaceutically acceptable excipient. 
     
     
         18 . A process according to  claims 17 , which comprises the further step of filling the antigen specific T cell population or a pharmaceutical composition comprising the same into a container such as infusion bag and sealing the container. 
     
     
         19 . An antigen-specific T cell population (such as an allogeneic antigen specific T cell population) obtained or obtainable from a process defined in any one of  claims 1  to  16 . 
     
     
         20 . An antigen specific T cell population (such as an allogeneic antigen specific T cell population) according to  claim 19 , wherein no more than 30 per cent of the cell are CD3−. 
     
     
         21 . An antigen specific cell population (such as an allogeneic antigen specific T cell population) according to  claim 19  or  20 , wherein the expansion is biased towards CD4+ cell expansion. 
     
     
         22 . An antigen specific T cell population (such as an allogeneic antigen specific T cell population) according to  claim 21 , wherein the CD4+ T cell population is primarily a Th1 population. 
     
     
         23 . A pharmaceutical composition expanded antigen-specific T cell population for a patient, wherein the composition comprises: a population of at least 70% CD3+ T cells and less than 30 percent of non CD3+ cells, where the total population of CD3+ cells does not exceed 5×10 4  cells per kg of patient, and said population of CD3+ cells comprises a therapeutic T cell population of CD4+ cells which have a primary characterization as Th1 characterised in that the relevant expanded antigen specific T cell population is able to expand in vivo. 
     
     
         24 . A pharmaceutical composition according to  claim 23  adapted for infusion. 
     
     
         25 . A pharmaceutical composition according to any one of  claims 1  to  4 , wherein expanded antigen specific T cell population is at least 1% per cent of the total cell population. 
     
     
         26 . A pharmaceutical composition according to any one of  claims 23  to  25 , wherein the expanded antigen specific T cell population is at least 5,000 cells, for example 10,000. 
     
     
         27 . A pharmaceutical composition according to any one of  claims 23  to  26 , wherein the expanded antigen-specific T cell population is in the range 20 to 80 per cent of the CD3+ T cell population. 
     
     
         28 . A pharmaceutical composition according to  claims 23  to  27 , wherein the expanded antigen specific T cell population is specific for a cell infected with CMV and/or presenting CMV antigens. 
     
     
         29 . A pharmaceutical composition according to any one of  claims 23  to  27 , wherein the expanded antigen specific T cell population is specific for a cell infected with adenovirus and/or presenting adenovirus antigens. 
     
     
         30 . A pharmaceutical composition according to any one of  claims 23  to  29 , wherein the expanded antigen specific cell population of the composition is expanded from a mobilised blood sample. 
     
     
         31 . A pharmaceutical composition according to any one of  claims 23  to  28 , wherein the cell population is cultured for 14 days or less, such as 12, 11 or 10 days. 
     
     
         32 . A pharmaceutical composition as defined in any one of  claims 23  to  31  for use in the treatment or prophylaxis of a patient, for example a patient who has received a HS Cell transplantation. 
     
     
         33 . A pharmaceutical composition according to  claim 32 , wherein the treatment or prophylaxis is for viral infection, for example CMV or ADV. 
     
     
         34 . A method of treating a patient comprising administering a therapeutically effective amount of amount of antigen specific T cell population as defined in  claims 19  to  22  or a pharmaceutical composition according to any one of  claims 23  to  31 .