The present invention relates to methods for developing engineered T-cells for immunotherapy that are non-alloreactive. The present invention relates to methods for modifying T-cells by inactivating both genes encoding T-cell receptor and an immune checkpoint gene to unleash the potential of the immune response. This method involves the use of specific rare cutting endonucleases, in particular TALE-nucleases (TAL effector endonuclease) and polynucleotides encoding such polypeptides, to precisely target a selection of key genes in T-cells, which are available from donors or from culture of primary cells. The invention opens the way to standard and affordable adoptive immunotherapy strategies for treating cancer and viral infections.

The present invention relates to the field of immunology, molecular biology and therapeutics. In particular, the invention relates to novel artificial feeder cells for activation and expansion of natural killer (NK) cells. The artificial feeder cell expresses endogenous ligands (HLA C1, C2, 5 and Bw4 type) for killer cell immunoglobulin-like receptors (KIRs), non-KIR binding Bw6 ligand, endogenous HLA-E-ligand for inhibitory NKG2A receptor, and comprises at least one stimulatory cytokine either membrane bound or secreted or at least one co-stimulatory ligand where those ligands and cytokines each specifically bind to a cognate receptor on a NK cell of interest, thereby mediating expansion of the NK cell. The invention can be used as an “off the 10 shelf” artificial feeder cell that can be readily designed to expand a NK cell or a NK subset of interest and also specifically expand NK cells modified with a chimeric antigen receptor (CAR). By genetically introducing or knockdown of candidate genes, the artificial feeder cell of the invention can be used to identify the stimulatory, co-stimulatory, and any other factors that mediate growth, expansion and cytotoxicity of a NK cell. Thus, the present invention provides 15 powerful tools for development of novel therapeutics where activation and expansion of the NK cell and of the CAR-NK cell can provide a benefit.

Methods and compositions for treating cancer in a subject in need thereof. The method includes administering to the subject an effective amount of a composition comprising Tumor-Targeting Effectors (TITE) derived from a culture comprising a bispecific antibody armed activated T cell (BAT) and a cancer cell, to thereby treat cancer in the subject.

The present invention relates to a modified natural killer (NK) cell and its use in personalised medicine. The modified NK cells of the present invention are non-immunogenic, meaning that they are able to be administered to any recipient subject without being rejected by the host immune system (they are “universal”). In a first embodiment the non-immunogenic NK cells are modified to express CD3 to allow a T-cell Receptor (TcR) to be expressed. In a further embodiment the non-immunogenic NK cells are further modified to express a TcR together with the CD3 co-receptor. Co-expression of CD3 with a specific TcR results in the modified NK cells showing antigen-specific cytotoxicity towards target cells. Universal NK cells can thus be targeted against specific antigens, and may thus be used in personalised medicine, particularly in the field of oncology.

The antigen-presenting cell loaded with the cancer-specific tumor antigen epitope provided in the present invention, that is, a dendritic cell enables rapid and effective induction of differentiation and proliferation of cancer antigen-specific T cells, preferably memory T cells, and the memory T cells thus activated can treat a cancerous or neoplastic condition or prevent recurrence, progression, or metastasis of cancer while avoiding the defense mechanism of cancer cells.

Several embodiments disclosed herein relate to methods and compositions for enhanced expansion of NK cells in culture. In several embodiments, the methods utilize one or more soluble interleukins as culture media supplements at one or more time points during expansion of the NK cell, or other immune cell, the expansion employing a feeder cell population.

Provided is a three-dimensional tissue, including: a first cellular region including cells of a first type; and a second cellular region including cells of a second type different from the first type, wherein the cells of the first type are cells that emit light by chemiluminescence, bioluminescence, or fluorescence in response to an external stimulus.

Described herein a polycistronic expression cassettes and expression vectors that include a promoter operably linked to a nucleic acid segment that encodes a Sox2 and Klf4 polypeptide. The nucleic acid segment can also encode a c-Myc polypeptide. Expression of such polycistronic expression cassettes/vectors in host cells can reprogram the host cells to stem cells or other types of reprogrammed cells.

Provided are modified microorganisms, such as live recombinant commensal bacteria, that express a heterologous antigen, and methods of using the modified microorganisms to induce an antigen-specific immune response to the heterologous antigen. The modified microorganism can be used to induce a regulatory T cell immune response to the heterologous antigen to treat an autoimmune disease in a subject in need thereof, or can be used to induce an effector T cell immune response to the heterologous antigen to treat a proliferative disease in a subject in need thereof.

The present invention relates to methods for developing engineered T-cells for immunotherapy and more specifically to methods for modifying T-cells by inactivating at immune checkpoint genes, preferably at least two selected from different pathways, to increase T-cell immune activity This method involves the use of specific rare cutting endonucleases, in particular TALE-nucleases (TAL effector endonuclease) and polynucleotides encoding such polypeptides, to precisely target a selection of key genes in T-cells, which are available from donors or from culture of primary cells. The invention opens the way to highly efficient adoptive immunotherapy strategies for treating cancer and viral infections.