A preparation and an application of a chimeric antigen receptor immune cell constructed on the basis of granzyme B are provided, including, a granzyme B (GrB)-based modified chimeric antigen receptor (CAR), the CAR containing an extracellular binding domain, and the extracellular binding domain being capable of specifically targeting heat shock proteins. The CAR immune cell acts by means of binding of a ligand and a receptor, and has relatively high targeting specificity and relatively better safety.

The present invention is directed to the field of immunotherapy. Specifically, the invention provides improved cell compositions and methods for adoptive cell therapy, useful in the treatment of cancer. More specifically, embodiments of the invention employ the use of cell compositions comprising a high proportion of activated cytotoxic CD8.sup.+ cells and in particular CD8.sup.+NKG2D.sup.+granzyme-B.sup.+ cells characterized by enhanced cytotoxicity, to processes for their preparation from peripheral blood mononuclear cells (PBMC), and to their use in cancer management.

Cell-targeted serine protease constructs are provided. Such constructs can be used in methods for targeted cell killing such as for treatment cell of proliferative diseases (e.g., cancer). In some aspects, recombinant serine proteases, such as Granzyme B polypeptides, are provided that exhibit improved stability and cell toxicity. Methods and compositions for treating lapatinib or trastuzumab-resistant cancers are also provided.

Cell-targeted serine protease constructs are provided. Such constructs can be used in methods for targeted cell killing such as for treatment cell of proliferative diseases (e.g., cancer). In some aspects, recombinant serine proteases, such as Granzyme B polypeptides, are provided that exhibit improved stability and cell toxicity. Methods and compositions for treating lapatinib or trastuzumab-resistant cancers are also provided.

The technology provided herein relates to novel immunoproteases suitable to induce apotosis in selected diseased target cells, comprising the serine protease granzyme M, and methods for using such cytolytic fusion proteins for the treatment of various diseases, in particular for the treatment of cancer.

Various embodiments disclosed relate to a nanoparticle-protein complex for intracellular protein delivery. In various embodiments, the present invention provides a nanoparticle-protein complex including a nanoparticle including an amine-containing ligand. The nanoparticle-protein complex also includes a protein comprising a carboxylic acid-containing tag.

The disclosure relates to a fusion protein comprising a colony stimulating factor 1 (CSF1) portion and a granzyme B portion, and polynucleotide sequence encoding the fusion protein. The disclosure also relates to a virus comprising the polynucleotide, and a T cell expressing the polynucleotide. The disclosure further relates to a method of treating a disease in an individual by administering the fusion protein, the polynucleotide, the oncolytic virus, or the T cell, and to a composition for use in method.

The invention is directed to methods and compositions for cell-based targeted delivery of predetermined compounds to a population of target cells. In some embodiments, methods of the invention include providing cytotoxic lymphocytes genetically modified to produce and sequester in lytic granules fusion proteins comprising a granzyme, or other effector agent, and a predetermined protein, so that upon specific contact of the cytotoxic lymphocytes with the target cells, the granzyme-perforin pathway of the cytotoxic lymphocytes is activated, leading to the delivery of the fusion protein to the cytosols of the target cells.

Disclosed herein are methods, systems and devices to model adaptive immune responses and develop and/or test improved antibodies, vaccines, and other therapeutic agents. The adaptive immune responses can be modeled using lymphoid tissue derived from a subject. The methods, systems and devices disclosed herein can comprise modified T-cells.

A polypeptide having a serine protease variant of the human granzyme B set forth by SEQ ID NO: 1, wherein the serine protease variant has at least 95% identity to SEQ ID NO: 1 and has a substitution at the position that corresponds structurally or by amino acid sequence homology to position Arg201 of SEQ ID NO: 1, and wherein the serine protease variant has activity to cleave the motif Ile-Glu-Thr-Asp.