9 results about "Granzyme" patented technology

Abstract Described herein, in one aspect, are antigen presenting cells (APCs) comprising an exogenous nucleic acid encoding one or more candidate antigens, wherein the one or more candidate antigens are expressed and presented with MHC class I or MC class II molecules; a molecular reporter of Granzyme B (GzB) activity; and c) an exogenous inhibitor of caspase-activated deoxyribonuclease (CAD)-mediated DNA degradation, a CAD knockout, or a caspase knockout (e.g., caspase 3 knockout). Described herein, in another aspect, is a system for detection of recognized antigen presentation by an antigen presenting cell to a cytotoxic lymphocyte or NK cell. Abstract 2018 / 22761 oM - cell Target ml Target cell cell cell Target Target Target SUBSTITUTE SHEET (RULE 26) cell cell cell Target Target cell cell 1 / 28 my Isolate recognized cell Library of target cells target cells and displaying different Add T cells from sample sequence antigens antigens of interest. CTLs deliver cytotoxic granules to target cells displaying cognate antigen FIG. 1 PCT / US2018 / 036663 20 26 20 53 56 07 J ul 2 02 6 2 0 2 6 2 0 5 3 5 6 0 7 J u l 2 0 2 6 2 0 1 8 / 2 2 7 6 1 o M a n d m y 1 / 2 8 m y L i b r a r y o f t a r g e t c e l l s d i s p l a y i n g d i f f e r e n tA d d T c e l l s f r o m s a m p l e of interest. CTLs deliver c y t o t o x i c g r a n u l e s t o t a r g e t c e l l s d i s p l a y i n g c o g n a t e a n t i g e n P C T / U S 2 0 1 8 / 0 3 6 6 6 3

Compositions and methods are provided for isolating, manipulating and using for therapeutic and other purposes, mammalian, MHC Class I restricted, antigen-specific regulatory T cells. The regulatory T cells can be characterized as CD8+ cells that specifically suppress the responses of self-reactive and / or pathogenic CD4+ T cells by cytotoxic mechanisms including, without limitation, perforin, other components of the perforin / granzyme apoptosis pathway, etc. The regulatory T cells are antigen-specific, but are not activated by the same antigen as the self-reactive and / or pathogenic CD4+ T cells. In humans the regulatory T cells express inhibitory KIR proteins, e.g. one or more of KIR2DL2, KIR2DL3, and KIR3DL1.

This invention relates to a granzyme B-targeting complex, a radiopharmaceutical, and their imaging applications in tumor immunotherapy, belonging to the field of nuclear medicine diagnostics. The complex is covalently modified, triggering a covalent group proximity effect by recognizing the binding of a peptide to granzyme B, achieving irreversible covalent linkage between the probe and the target protein. This significantly enhances the probe's selective retention at the target site and extends the imaging time window. After radionuclide labeling, the complex can be prepared into a radiopharmaceutical for nuclear medicine imaging; the labeling process is simple and exhibits good stability. This radiopharmaceutical allows for non-invasive, specific, and quantitative monitoring of granzyme B expression levels via PET or SPECT, reflecting the activity status of immune cells in vivo. This invention has significant clinical application value and promising prospects for the early and accurate prediction and dynamic monitoring of tumor immunotherapy efficacy.

This invention discloses a method and application for single-cell nucleus dissociation suitable for tissue sections. The method includes: S1, obtaining tissue sections to be processed, attaching them to a carrier surface, and then adding an incubation buffer for incubation; wherein the incubation buffer contains Tris-HCl, NaCl, MgCl2, micro / nano particles, RNase inhibitors, bovine serum albumin, and a nonionic surfactant; S2, transferring the incubated product to a centrifuge tube, gently mixing by pipetting to obtain a nucleus dissociation buffer; S3, filtering and centrifuging the nucleus dissociation buffer, collecting the precipitate, and resuspending it to obtain a nucleus suspension. This single-cell nucleus dissociation method is mild and efficient. The nucleus suspension obtained using this method has high purity of single nuclei and low cytoplasmic debris, and can be directly used for downstream omics analyses such as high-throughput single-cell nuclear sequencing, spatial transcriptomics, and ATAC-seq, demonstrating high application value.

The application discloses a fluorescence hybridization chain reaction biosensor based on the existence of granzyme B and application thereof, and realizes real-time detection of T cell effector molecules by establishing a fluorescence hybridization chain reaction biosensor based on the existence of active granzyme B, and has the characteristics of short amplification time, simple operation, high amplification efficiency, simple reaction system and high economic benefit. By introducing peptide nucleic acid into the hybridization chain reaction system, the cutting of the substrate by granzyme B is successfully used as the starting recognition event of the hybridization chain reaction, and after the cutting of the peptide substrate in the peptide nucleic acid by granzyme B, the foothold is exposed, the strand displacement reaction is mediated, the fluorescence signal is restored after amplification, and a new strategy of using the hybridization chain reaction for detecting active cytokines is provided. The strategy makes the hybridization chain reaction using fluorescence as a signal output mode applicable to active protein detection, and has the advantages of a simple reaction system, few operation steps and low detection cost.

Compounds that bind to granzyme B and can contain a radioactive moiety, such as the compound of formula (I), and pharmaceutical compositions comprising the same. Uses of the compound and pharmaceutical composition in cancer treatment, as well as kits and methods for cancer treatment, are also provided herein.

A kind of granzyme B targeting complex, radiopharmaceutical and its imaging application in intestinal immune abnormal activity belong to the field of nuclear medicine diagnosis and treatment.The complex is covalently modified, specifically non-covalently combined with granzyme B by recognizing peptide first, and then triggers irreversible covalent connection by means of proximity effect, effectively solves the technical bottleneck of low target tissue uptake, short in vivo retention time and narrow imaging window of existing granzyme B probe, significantly improves the selective retention capacity and imaging specificity of the probe in the target site.The complex and radiopharmaceutical of the present application can specifically target granzyme B released by intestinal lesion site immune abnormal activity, realize non-invasive, dynamic, quantitative nuclear medical imaging of intestinal inflammation such as Crohn's disease, ulcerative colitis and aGVHD, and accurately evaluate the degree of intestinal immune abnormal activity and monitor the treatment effect, providing a core tool for early diagnosis and precise diagnosis and treatment of such diseases, and having important clinical transformation value and application prospect.

This invention discloses an NK cell exosome capable of targeting and killing lung cancer and its preparation method. Specifically, it is based on NK cells derived from umbilical cord blood stem cells, and involves constructing CRISPR-Cas9-PD-1 knockout recombinant plasmids to transfect cells, and rapidly inducing umbilical cord blood stem cells to transform into CD3+ cells using IL-21. ‑ CD56 + NK cell differentiation and maturation, tandem ultracentrifugation, and size exclusion chromatography were used for high-efficiency purification to obtain PD-1 knockout NK cell exosomes containing granzyme, perforin, and FasL. L NK-EXO). The PD-1 constructed in this invention L NK-EXO treatment significantly prolonged the survival of a mouse model of lung cancer and reduced tumor size, while also demonstrating clear lung cancer targeting and specific killing functions. More importantly, this invention reveals that PD-1 knockout of NK cell exosomes is crucial for tumor targeting and killing.