JPWO2018211115A5 - - Google Patents

Description

Sources of immune effector cells can include both allogeneic and autologous sources. In some cases, immune effector cells can be differentiated from stem cells or induced pluripotent stem cells (iPSCs). Thus, cells for manipulation according to this embodiment can be isolated from cord blood, peripheral blood, human embryonic stem cells or iPSCs. For example, allogeneic T cells can be modified to contain a chimeric antigen receptor (and optionally lack a functional TCR). In some aspects, the immune effector cells are primary human T cells, such as T cells derived from human peripheral blood mononuclear cells (PBMC). PBMC can be harvested from peripheral blood or after stimulation with G-CSF (granulocyte colony stimulating factor) from bone marrow or cord blood. After transfection or transduction (eg, using a CAR expression construct), cells can be injected immediately or can be cryopreserved. In some embodiments, the cells are grown ex vivo as a bulk population for days, weeks, or within about 1, 2, 3, 4, 5 days or more after transfection and after introduction of the gene into the cells. It can be grown for several months. In another embodiment, after transfection, transfectants are cloned and clones exhibiting the presence of integrated or episomally maintained single expression cassettes or plasmids and expression of chimeric antigen receptors ex vivo. grow in Clones selected for expansion demonstrate the ability to specifically recognize and lyse antigen-expressing target cells. The recombinant T cells can be expanded by stimulation with IL-2 or other cytokines that bind to the common gamma chain (eg, IL-7, IL-12, IL-15, IL-21, etc.). The recombinant T cells can be expanded by stimulation with artificial antigen presenting cells. The recombinant T cells can be grown on artificial antigen presenting cells or using an antibody such as OKT3 that cross-links CD3 on the T cell surface. The recombinant T cell subsets can be further selected using magnetic bead-based isolation methods and/or fluorescence-activated cell sorting techniques and further cultured in the presence of AaPCs. In another aspect, genetically modified cells can be cryopreserved.

Claims (12)

1. A pharmaceutical composition for use in the treatment of infection or cancer comprising a polypeptide construct that selectively binds to the J configuration of CD277 on a target cell, said polypeptide construct comprising a γ9δ2 T cell receptor, said A pharmaceutical composition wherein the polypeptide construct is soluble and the polypeptide construct also binds to cytotoxic T cells and activates the cytotoxic T cells to be cytotoxic to target cells . The pharmaceutical composition for use according to claim 1, wherein CD277 exists as a dimer. 2. A pharmaceutical composition for use according to claim 1, wherein the polypeptide construct binds to the J configuration of CD277 with increased selectivity relative to CD277 molecules that are not in the J configuration. The pharmaceutical composition for use according to claim 1, wherein said cancer is leukemia or solid cancer. The pharmaceutical composition for use according to any one of claims 1 to 4, wherein said subject has CD277-positive cancer cells. A mevalonate pathway inhibitor further comprising administering a dosage form of an agent that enhances the activity of RhoB GTPase in cancer cells and an agent that enhances the activity of phosphoantigen in cancer cells, wherein the agents are aminobisphosphonates. A pharmaceutical composition for use according to any one of claims 1 to 5, which is 7. A pharmaceutical composition for use according to claim 6, wherein formation of the J configuration requires at least interaction of RhoB with CD277 and/or compartmentalization of CD277. 8. A pharmaceutical composition for use according to claim 7, wherein formation of the J configuration requires interaction of intracellular phosphoantigen with CD277 after interaction of RhoB with CD277. 1. A method of predicting a positive therapeutic response in a subject to treatment with a polypeptide construct capable of recognizing CD277 or treatment with an engineered cell expressing said polypeptide construct, comprising:
the polypeptide construct contains a γ9δ2 T cell receptor;
The method comprises steps a) and b) below.
a) identifying a target cell of the patient as having a nucleotide sequence polymorphism associated with the activity of RhoB; and b) demonstrating a positive therapeutic response based on said identification of the target cell as having a nucleotide sequence polymorphism. predicting the target;
The method further comprising predicting poor therapeutic response in the second subject based on identifying target cells of the second subject as lacking the nucleotide sequence polymorphism. 10. The method of claim 9, wherein said polypeptide construct recognizes the J configuration of CD277. 10. The method of claim 9, wherein one or more of said target cells are cancer cells. 10. The method of claim 9, wherein the RhoB activity is elevated compared to RhoB activity in a subject lacking the nucleotide sequence polymorphism.