Platform for activation and expansion of virus-specific t-cells

Abstract

Embodiments of the disclosure concern methods and compositions for immunotherapy for diseases and malignancies associated with viruses other than HPV or with non-virus-associated diseases and malignancies, such as wherein the VST encodes a CAR specific for a non-viral cancer and the VST can be stimulated in vitro or in vivo using viruses, viral vaccines or oncolytic viruses. In specific embodiments, methods concern production of immune cells that target one or more antigens of HIV, EBV, CMV, adenovirus, vaccinia virus, and/or VZV, including methods with stimulation steps that employ IL-7 and IL-15, but not IL-2, IL-4, or both. Other specific embodiments utilize stimulations in the presence of certain cells, such as costimulatory cells and certain antigen presenting cells.

1 . A method of producing viral antigen-specific T-cells, comprising the step of stimulating a population of peripheral blood T-cells with antigen presenting cells in the presence of IL-7 and IL-15, wherein the antigen presenting cells are or were previously exposed to a library of peptides, wherein the peptides comprise sequence that corresponds to at least part of the sequence of one or more proteins of one or more viruses that are not HPV, wherein the method comprises one or more of the following:
 (a) the method occurs in the absence of IL-4;   (b) the concentration of IL-15 is ≥100 ng per mL; and   (c) the method comprises depletion of CD45RA-positive cells from the peripheral blood T-cells   
     
     
         2 . The method of  claim 1 , wherein the stimulating occurs in the absence of IL-6, IL-12, IL-2, IL-21, or a combination thereof. 
     
     
         3 . The method of  claim 1 , wherein the population of peripheral blood T-cells have reduced levels, compared to normal levels, of one or more of the following: 1) NK cells; 2) naïve cells that can grow as bystander cells; and/or 3) T-regulatory cells. 
     
     
         4 . The method of  claim 3 , wherein peripheral blood mononuclear cells (PBMCs) or peripheral blood T-cells obtained therefrom are subject to a step of reducing the levels of one or more of the following: 1) NK cells; 2) naïve cells that can grow as bystander cells; 3) T-regulatory cells and/or 4) inhibitory myeloid cells. 
     
     
         5 . The method of  claim 1 , wherein the virus is from the family Herpesviridae or is a poxvirus, adenovirus, polyomavirus, lentivirus, rhabdovirus or other oncolytic virus. 
     
     
         6 . The method of  claim 1 , wherein the virus is selected from the group consisting of Epstein-Barr virus (EBV), Cytomegalovirus (CMV), adenovirus, vaccinia virus, and/or Varicella zoster virus (VZV), HIV, influenza, marabavirus vesicular stomatitis virus and an oncolytic virus. 
     
     
         7 . The method of  claim 1 , wherein the peripheral blood T-cells are present in a population of PBMCs or are obtained or isolated therefrom. 
     
     
         8 . The method of  claim 4 , wherein the PBMCs or apheresis product are depleted of one or more of the following: 1) NK cells; 2) naïve cells that can grow as bystander cells; and/or 3) T-regulatory cells. 
     
     
         9 . The method of  claim 4 , wherein the PBMCs in the population are non-adherent PBMCs 
     
     
         10 . The method of  claim 1 , wherein the antigen presenting cells are dendritic cells or PBMCs. 
     
     
         11 . The method of  claim 1 , wherein a stimulating step occurs in the presence of costimulatory cells. 
     
     
         12 . The method of  claim 11 , wherein the costimulatory cells are CD80+, CD86+, CD83+, 4-1BBL+, or a combination thereof, or wherein the costimulatory cells are HLA-negative lymphoblastoid cells. 
     
     
         13 . The method of  claim 1 , wherein the stimulating occurs in the presence of activated T-cells, dendritic cells, PBMCs, or HLA-negative costimulatory cells. 
     
     
         14 . The method of  claim 13 , wherein when the stimulating occurs in the presence of activated T-cells, dendritic cells, PBMCs, or HLA-negative costimulatory cells, the stimulating step is not a first stimulation step. 
     
     
         15 . The method of  claim 13 , wherein the activated T-cells are autologous to the individual. 
     
     
         16 . The method of  claim 1 , wherein the stimulating occurs in the presence of pepmixes, pepmix-pulsed autologous activated T-cells, in the presence of HLA-negative costimulatory cells, or both. 
     
     
         17 . The method of  claim 16 , wherein when the stimulating occurs in the presence of pepmix-pulsed autologous activated T-cells, in the presence of HLA-negative costimulatory cells, or both, the stimulating step is not a first stimulation step. 
     
     
         18 . The method of  claim 1 , wherein the peptide library comprises peptides of at least or no more than 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more amino acids in length. 
     
     
         19 . The method of  claim 1 , wherein the peptide library comprises peptides of 15 amino acids in length. 
     
     
         20 . The method of  claim 1 , wherein peptides in the library overlap in sequence with other peptides by 11 amino acids. 
     
     
         21 . The method of  claim 1 , wherein T-cells produced by a first stimulating step are subject to one or more subsequent stimulating steps. 
     
     
         22 . The method of  claim 21 , wherein a subsequent stimulating step occurs in the presence of IL-7 and IL-15. 
     
     
         23 . The method of  claim 21 , wherein a subsequent stimulating step occurs in the presence of activated T-cells, costimulatory cells, IL-7, and IL-15. 
     
     
         24 . The method of  claim 1 , wherein the method occurs in the absence of exposing the T-cells produced by the method to activated B cells that were previously exposed to a library of peptides. 
     
     
         25 . The method of  claim 1 , wherein the cells are modified to express a gene product from an expression vector. 
     
     
         26 . The method of  claim 25 , wherein the cells are modified to express a chimeric antigen receptor, αβT-cell receptor, or combination thereof. 
     
     
         27 . The method of  claim 1 , wherein a therapeutically effective amount of T-cells produced by the method are provided to an individual that has been exposed to EBV, CMV, adenovirus, vaccinia virus, HIV, and/or VZV, that is seropositive for EBV, CMV, adenovirus, vaccinia virus, HIV, and/or VZV, or that has disease associated with EBV, CMV, adenovirus, vaccinia virus, HIV, and/or VZV. 
     
     
         28 . The method of  claim 1 , wherein the individual is determined as having a medical condition associated with EBV, CMV, adenovirus, vaccinia virus, HIV, and/or VZV. 
     
     
         29 . The method of  claim 1 , wherein one or more steps of the method lack the presence of exogenously added IL-4, IL-2, or both. 
     
     
         30 . A method for stimulating T-cells specific for a virus other than HPV, comprising stimulating T-cells specific for the virus with antigen presenting cells in the presence of IL-7 and IL-15 but in the absence of IL-4 and in the presence of co-stimulatory cells, wherein the antigen presenting cells were previously exposed to one or more peptides, wherein the peptides comprise sequence that corresponds to at least part of the sequence of one or more proteins of a virus other than HPV, and wherein the antigen presenting cells are depleted of CD45RA-positive cells. 
     
     
         31 . A method of producing therapeutic T-cells for a virus-associated disease or a non-virus-associated disease, the method comprising the step of stimulating T-cells specific for a virus other than HPV with antigen presenting cells in the presence of one or more of IL-7 and IL-15 but in the absence of IL-4 and in the presence of co-stimulatory cells, wherein the antigen presenting cells were previously exposed to one or more peptides, wherein the peptides comprise sequence that corresponds to at least part of the sequence of one or more proteins of a virus other than HPV, wherein the stimulating produces T-cells therapeutic for the virus-associated disease, wherein the antigen presenting cells are depleted of CD45RA-positive cells.