The present disclosure relates generally to culture media formulations and culture methods. More particularly, the present disclosure provides defined serum-free culture media, kits and methods for generating progenitor T cells and derivatives thereof, including mature T cells. The present disclosure further provides the cells generated using the media, kits and methods, as well as methods of treatment using the generated cells.

Provided are NK-92 cells expressing a chimeric antigen receptor (CAR). The CAR can comprise an intracellular domain of FcϵRIγ. Also described are methods for treating a patient having or suspected of having a disease that is treatable with NK-92 cells, such as cancer or a viral infection, comprising administering to the patient NK-92-CAR cells.

Provided is an antibody that specifically recognizes IL-13RA2, which can be used in the manufacture of a targeting anti-tumor medicament as well as a medicament for diagnosing a tumor.

Embodiments of the disclosure encompass methods and compositions for producing engineered natural killer (NK) cells. The disclosure concerns large-scale processes for producing NK cells that are engineered to have disruption of expression of one or more genes, such as using CRISPR, and also engineered express at least one heterologous antigen receptor. Specific embodiments include particular parameters for the process.

In order to improve the efficiency of gene introduction in CAR therapy employing a transposon method, provided is a method for preparing genetically-modified T cells expressing chimeric antigen receptor, comprising: (1) a step of preparing non-proliferative cells which are obtained by stimulating a group of cells comprising T cells using an anti-CD3 antibody and an anti-CD28 antibody followed by a treatment for causing the cells to lose their proliferation capability; (2) a step of obtaining genetically-modified T cells into which a target antigen-specific chimeric antigen receptor gene has been introduced using a transposon method; (3) a step of mixing the non-proliferative cells prepared by step (1) with the genetically-modified T cells obtained by step (2), and co-culturing the mixed cells while stimulating the mixed cells using an anti-CD3 antibody and anti-CD28 antibody; and (4) a step of collecting the cells after culture.

A method for coupling an antibody to a surface of a cell comprises the following steps: (1) chemically modifying sialic acid to obtain a sialic acid derivative containing an azide group; (2) absorbing the sialic acid derivative by the cell to obtain a cell modified with the azide group; (3) modifying the antibody with a conjunction compound to obtain a modified antibody; (4) co-culturing the modified antibody with the cell modified with the azide group. The present disclosure modifies natural sialic acid molecules in vitro through chemical synthesis methods, and utilizes modified natural sialic acid molecules to realize antibody modification on the surface of the cell. The modification method of the present disclosure is simple, low-cost, safe, and efficient, does not need complex gene editing or enzyme catalytic operation, and has universality. In theory, the modification method can realize the coupling of any antibody or macro-molecular substance on the surface of the cell.

Embodiments of the disclosure provide methods and compositions that facilitate cancer treatment including at least because they concern therapies that circumvent the tumor microenvironment. In specific embodiments, compositions are utilized for therapy that utilize NK cells that are protected from the direct inhibition of their activity (using TGF-beta inhibitors) and/or that are indirectly protected from TGF-beta (using integrin inhibitors). In specific embodiments, the NK cells have deficient expression and/or activity for TGF-beta Receptor 2 and/or glucocorticoid receptor.

Disclosed herein are invariant natural killer T (iNKT) cells engineered using hematopoietic stem and progenitor cells (HSPCs) and methods of making and using thereof.

The invention relates to: a composition for culturing natural killer cells, comprising, as an active ingredient, a fusion protein comprising an IL-2 protein and a CD80 protein; and a method for preparing natural killer cells by using same. Particularly, a composition for culturing natural killer cells, comprising, as an active ingredient, a fusion protein comprising IL-2 or a variant thereof and CD80 or a fragment thereof, of the present invention; promotes the proliferation of natural killer cells, induces the expression of CD16 and NKp46, and increases the expression and secretion of granzyme B and perforin, thereby being effectively usable in the preparation of natural killer cells having an excellent anticancer immune function.

Certain universal neoepitopes and cancer specific neoepitopes and methods therefore are presented that may be used in immunotherapy and cancer diagnosis. Preferred therapeutic and diagnostic compositions include antibodies or fragments thereof that bind to neoepitopes on cancer cells.