This document provides methods and materials for generating T cells expressing a selected antigen receptor (e.g., a chimeric antigen receptor) in vivo. For example, viral particles containing nucleic acid encoding a selected antigen receptor, cells capable of producing such viral particles, and methods for administering such viral particles and/or cells to a mammal to generate T cells expressing that selected antigen receptor within the mammal are provided.

Disclosed herein are genome-edited chimeric antigen receptor T cells (CAR-T), which can be derived from a cytotoxic T cells, a viral-specific cytotoxic T cell, memory T cells, or gamma delta () T cells, and comprise one or more chimeric antigen receptors (CARs) targeting one or more antigens, wherein the CAR-T cell is deficient in one or more antigens to which the one or more CARs specifically binds. In particular, the present disclosure relates to engineered mono, dual, and tandem chimeric antigen receptor (CAR)-bearing T cells (CAR-T) and methods of immunotherapy for the treatment of cancer.

The present invention is defined by the claims. In particular, the present invention relates to the use of TCR-deficient CAR-Tregs in combination with anti-TCR complex monoclonal antibodies for inducing durable tolerance.

Provided are methods of continuous counterflow centrifugation for the manufacturing of cell compositions, including for the production of T cell therapies including cells that express recombinant receptors such as chimeric antigen receptors (CARs).

A preparation method for a universal CAR-T cell, and an use of a universal CAR-T cell. When universal CAR-T is prepared, an RNP complex comprises an RNP complex composed of a Cas9 protein and sgRNA; and the SgRNA is SgRNA of CD4 and/or sgRNA of CD8. he CD4 gene and/or CD8 gene of the universal CAR-T cell or a T cell are/is knocked out and/or inactivated. The CAR-T cell expresses CD44 and/or CD62L, so that the activity of the CAR-T cell can be enhanced, the effectiveness of CAR-T is enhanced, the CAR-T cell has a better phenotype and lower exhaustion expression, and the killing of the CAR-T cell against a tumor target cell is improved, and thus, the CAR-T cell can be used for targeted therapy of tumors and can be used for allogenic CAR-T therapy.

The present disclosure pertains to a method of construction and use of a novel bispecific chimeric antigen receptor (CAR) within the field of immunotherapy. The CAR described herein comprises an antigen-binding domain, a connecting peptide (C-peptide), a hinge region, a transmembrane domain, a 4-1BB co-stimulatory signaling domain, and a CD3 signaling domain. The antigen-binding domain is composed of either an anti-CD19 scFv and an anti-CD22 nanobody, or an anti-Her2 scFv paired with a ligand capable of recognizing IGF1R. The bispecific CAR-T cell provided by the present disclosure is constructed based on the ligation design involving an antiparallel -stranded loop (BS Loop) linker.

Provided is a set of viral vectors, comprising: a first viral vector that carries a first nucleic acid molecule encoding an envelope protein, and a second viral vector that carries a second nucleic acid molecule encoding a fusion protein, the fusion protein comprising: a single-chain antibody capable of binding to CD28 or CD3, and a C-terminal domain, comprising a transmembrane region and an intracellular region, of the envelope protein. The C terminal of the single-chain antibody is connected to the N terminal of the C-terminal domain of the envelope protein, and the envelope protein and the fusion protein are in a non-fusion form. Also provided are a method for obtaining a lentivirus and an obtained lentivirus, a method for introducing a lentivirus into an unactivated T lymphocyte, a method for expressing a target gene, a method for obtaining a CAR-T cell and an obtained CAR-T cell, a pharmaceutical composition, and a use thereof.

The present disclosure provides compositions and methods for engineering T cells (e.g., v1 T cells and v2 T cells) with an IL-15 receptor subunit (IL-15R), e.g., by transduction with a viral vector, to restore IL-15 responsiveness. Further provided are populations of engineered T cells and methods of using the same.

Provided herein are methods, uses, compositions, and kits that include hyaluronidase polypeptides, including for example retroviral particles that include membrane bound hyaluronidase polypeptides, and novel, chimeric hyaluronidase polypeptides. Furthermore, provided herein are methods, uses, compositions, and kits that include hyaluronidase polypeptides, including for example retroviral particles that encode hyaluronidases and/or that include membrane bound hyaluronidases, which in some embodiments are novel, chimeric hyaluronidases provided herein. Accordingly, provided herein are replication incompetent recombinant retroviral particles (RIP), and methods using the same, that comprise a viral envelope, a nucleic acid encoding a packaging signal, Gag, and a membrane-bound hyaluronidase.

Aspects of the invention described herein include methods of treating, inhibiting, ameliorating and/or eliminating a virus or cancer cells in a subject utilizing genetically engineered human T-cells having receptors for a molecule presented by the virus or the cancer cells, wherein the genetically engineered T cells are isolated utilizing a two-stage MTX selection that employs increasing concentrations of MTX.