48 results about "CD163" patented technology

The application belongs to the technical field of biotechnology, and particularly relates to a marker for prognosis evaluation of acute myeloid leukemia and application thereof, wherein the marker is HCK-positive mono-like cells, the HCK-positive mono-like cells are determined by co-positivity of antigens CD163, antigen CD68, antigen FCN1 and antigen HCK, and the application provides a marker with high sensitivity, strong specificity and good universality, which is used for detecting adult acute myeloid leukemia bone marrow or peripheral blood samples, so as to assist in judging the clinical prognosis of patients with acute myeloid leukemia.

The invention belongs to the technical field of biology, and discloses a BHK-21 cell line, the BHK-21 cell line can express porcine CD163 protein, the cell line is high in hereditary stability and high in PRRSV sensitivity, can be used in virus proliferation, vaccine production and basic experiments, and can provide a new tool for PRRSV vaccine production and mandatory research. Meanwhile, the invention also discloses a preparation method and application of the cell line.

The present application relates to a porcine reproductive and respiratory syndrome virus infectious clone construction and rescue method, and belongs to the technical field of bioengineering. The porcine reproductive and respiratory syndrome virus infectious clone contains a full genome sequence of the porcine reproductive and respiratory syndrome virus, and the nucleotide sequence of the full genome sequence is the 26th to 15384th nucleotide of SEQ ID NO: 1 or a fragment having more than 80% identity with SEQ ID NO: 1 and having the same function. The present application also provides a plasmid pCAGGS-HA-CD163 expressing a porcine CD163 gene. By co-transfecting the aforementioned two plasmids into HEK-293T cells, the virus is rescued in vitro, a first porcine reproductive and respiratory syndrome virus infectious clone rescue platform based on HEK-293T cells is successfully built, and the porcine reproductive and respiratory syndrome virus is successfully rescued.

The invention discloses a method for preparing a CD163 gene editing pig based on a CRISPR (clustered regularly interspaced short palindromic repeats) / Gs12-7MAX system. The method specifically comprises two editing strategies: one is to edit a sixth intron and a seventh intron of a CD163 gene to realize accurate deletion of a seventh exon so as to obtain a porcine fibroblast monoclonal cell with homozygous deletion of the seventh exon; and 2, editing aiming at a seventh exon of the CD163 gene, and inducing the exon to generate frame-shift mutation to obtain a gene editing pig. The technical system established by the invention has the advantages of high editing efficiency, strong accuracy and simplified operation process, provides a new effective strategy for prevention and control of PRRSV (porcine reproductive and respiratory syndrome), and has important popularization and application values in creation of new materials for pig disease-resistant breeding.

The present disclosure relates to methods of creating Porcine Reproductive and Respiratory Syndrome virus (PRRSv) resistant pigs by gene silencing. In particular, the methods comprise using tools such as CRISPR / CAS9 to edit genes that either directly encode or encode molecules that are processed into non-coding RNA molecules to induce or modify their silencing activity against CD163 and / or in cells of pigs for the purpose of promoting PRRSv resistance.

This invention discloses an epitope peptide of the key SRCR5 domain of porcine reproductive and respiratory syndrome virus (PRRSV) CD163, an epitope peptide vaccine, and its applications. The epitope vaccine of this invention exhibits good safety, without safety issues such as virulence reversion, viremia, or recombination with other PRRSV strains. It also demonstrates good immunogenicity; 14 days after secondary immunization, it can induce pigs to produce specific antibodies against the CD163 SRCR5-FP14 epitope with a maximum agglutination titer of 1:16. Immunized pigs can resist challenge from the highly pathogenic PRRSV JAX1 strain. Since all PRRSV subtypes rely on interaction with the CD163 receptor to infect susceptible host cells, and the CD163 molecule is conserved, the epitope vaccine blocking PRRSV infection also provides broad-spectrum protection against multiple subtypes without affecting pig production performance. This novel epitope vaccine has broad potential applications in PRRS prevention and control.

The application of the biomarker combination in the preparation of a drug for fulminant myocarditis belongs to the field of drugs. The application is characterized in that the biomarker combination comprises Siglec-5; the biomarker combination is further selected from the group consisting of sST2, PAI-1, Siglec-5, CD163, CD40, P-Cadherin, CD14, CTLA4. The application finds that the expression of the biomarker in the plasma of a patient with fulminant myocarditis is increased, and confirms the role of the biomarker as a drug target in relieving and / or improving fulminant myocarditis. The level of the biomarker can be controlled by a drug, the diagnosis and subsequent treatment of the disease can be more intensified, and the biomarker has great application value in clinical practice.

Provided herein are, inter alia, multispecific proteins that are, e.g., capable of simultaneously binding EpCAM (e.g., hEpCAM) and CD163 (e.g., hCD163) expressed on the surface of the same cell; nucleic acid molecules encoding the same; and cells expressing the same. The disclosure further relates to pharmaceutical composition comprising the same; and methods of utilizing the same, including, e.g., methods of treating cancer.

To provide pigs that exhibit resistance to porcine reproductive respiratory syndrome virus (PRRSV). [Solution] This disclosure relates to a genetically modified pig in which at least one allele of the SIGLEC-1 gene is inactivated and / or at least one allele of the CD163 gene is inactivated. A genetically modified pig in which both alleles of the SIGLEC-1 gene and / or both alleles of the CD163 gene are inactivated is resistant to porcine reproductive respiratory syndrome virus (PRRSV). Methods for producing such transgenic pigs are also provided.

The invention relates to the technical field of biological medicine and tissue engineering, in particular to a tissue engineering blood vessel and a preparation method and application thereof. The tissue engineering blood vessel is obtained by modifying interleukin 24 (IL-24) on the surface of an acellular blood vessel, after the tissue engineering blood vessel is transplanted into a body, the number of CD31 + and CD34 + cells can be remarkably increased, and infiltration of M2 type macrophages (CD163 +) on the surface of the blood vessel is remarkably increased; the expression of HIF-1 alpha (hypoxia marker) and MMP9 is obviously reduced; the expression of the nerve specific protein S-100 is obviously increased, thrombosis and intimal hyperplasia are effectively inhibited, and the patency rate of transplanted blood vessels is obviously increased.

The invention provides a monoclonal antibody for detecting pig source CD163 protein and a corresponding antigen epitope peptide. Wherein the hybridoma cell strain used for preparing the monoclonal antibody is CCTCC (China Center For Type Culture Collection) NO: C2025258. The monoclonal antibody provided by the invention is good in specificity, has good affinity with pCD163, and is clear in epitope information. The antibody can significantly inhibit the infection efficiency of PRRSV pedigree strains in alveolar macrophages, has a good PRRSV in-vitro blocking effect, can be used for developing a high-specificity pCD163 detection reagent, and is expected to be used as a candidate drug for passive immune prevention and treatment of PRRS.

The invention discloses a method for constructing MSTN (myostatin) gene knockout porcine primary cells, which comprises the following steps: designing and constructing plasmids capable of efficiently knocking out MSTN genes by utilizing an efficient and simple gene modification technology TALEN (transcription activator-like effector nodules) technology; a gene editing target site is designed for a pig MSTN gene, the target site can be specifically recognized by Cas9 endonuclease, so that double-strand breakage is mediated, the gene is mutated in a non-homologous end connection mode under the action of a self-repairing system, then an sgRNA composition is designed, tRNA-sgRNA-sgRNA scaffold is used as a minimum repetitive unit, and the target site is specifically recognized by Cas9 endonuclease. The Cas9-CD163-MSTN polycistron vector is constructed by using a PTG / Cas9 strategy, so that the target MSTN gene is efficiently and accurately knocked out, and therefore, the Cas9-CD163-MSTN polycistron vector has a relatively good application prospect in pig germplasm improvement.

The present disclosure provides a monoclonal antibody that binds to porcine CD163 for use in the treatment or prevention of porcine reproductive and respiratory syndrome (PRRS) virus infection in a pig. Preferred antibodies comprise an antigen binding domain that binds to porcine CD163, the antigen binding domain comprising at least one heavy chain variable region comprising three complementarity determining regions (CDRs), wherein the heavy chain variable region comprises a variable heavy chain (VH) CDR2 comprising the amino acid sequence XYAD or XYAE or XYAN, wherein X can be any amino acid. Also provided are nucleic acid molecules, expression vectors, and compositions.

The present disclosure provides compositions and methods for decreasing permissiveness to infection by PRRSV. The methods involve inserting peptides or oligopeptides into the CD163 SRCR domain 5 to disrupt the protein structure and thereby prevent PRRSV infection. The deletion of the exon 13 region of PSTII eliminates infection. The replacement of cysteine residues with alanines in SRCR5 as a means to disrupt disulfide bond formation. The final method is the deletion of, or amino acid substitution within the SRCR4-5 interdomain region.

The present disclosure relates methods and compositions useful for prevention of porcine reproductive and respiratory syndrome virus (PRRSv) in animals, including animals of the species Sus scrofa. The present teachings relate to swine wherein at least one allele of a CD163 gene has been inactivated, and to specific methods and nucleic acid sequences used in gene editing to inactivate the CD163 gene. Swine wherein both alleles of the CD163 gene are inactivated are resistant to porcine reproductive and respiratory syndrome virus (PRRSv). Elite lines comprising homozygous CD163 edited genes retain their superior properties.

This invention relates to improved antibodies targeting CD163. In particular, this invention relates to improved antibodies targeting CD163 with improved capabilities associated with inducing antibody-dependent cell-mediated cytotoxicity (ADCC).

This invention provides a monoclonal antibody against CD163, antibody combinations, and related applications. Utilizing single-B cell sequencing technology, this invention identifies 19 anti-CD163 antibodies belonging to multiple different antigen-binding epitope groups, offering greater antibody diversity and providing more possibilities for selecting paired antibodies. It is applicable to various scenarios such as ELISA, flow cytometry, proteoblotting, and immunohistochemistry.

The invention discloses a porcine reproductive and respiratory syndrome virus CD163 key SRCR5 structural domain epitope peptide, an epitope peptide vaccine and application. The epitope vaccine is good in safety, has no safety problems of virulence reversion, generation of viremia, recombination with other PRRSV strains and the like, has good immunogenicity, and can induce a pig to generate a specific antibody aiming at the CD163 SRCR5-FP14 epitope with the highest agglutination titer of 1: 16 14 days after secondary immunization, and the immunized pig can resist attack of a highly pathogenic PRRSV JAX1 strain. Each subtype strain of the PRRSV is dependent on interaction with a CD163 receptor so as to infect a host susceptible cell, and CD163 molecules are conservative, so that the epitope vaccine for blocking PRRSV infection also has a broad-spectrum protection effect on various subtype strains, the production performance of pigs is not influenced, and the novel epitope vaccine has a wide potential application prospect in PRRS prevention and control.

The invention discloses a method for efficiently enriching anti-blue ear and porcine reproductive capacity edited pig donor cells. The method comprises the following steps: introducing CRISPR-Cas9 plasmids targeting a BMP15 gene, CRISPR-Cas9 plasmids targeting a CD163 gene and alternative reporter plasmids edited by double genes into the pig donor cells; culturing the porcine donor cells to enable gene editing and reporter gene expression to occur; detecting and sorting cells with positive report gene expression so as to obtain enriched double-gene editing donor cells; the alternative reporter plasmid for double-gene editing comprises the following elements which are connected in sequence: a recognition sequence of a BMP15 gene editing target, a transcription terminator sequence, a recognition sequence of a CD163 gene editing target and a fluorescent reporter gene sequence. According to the invention, the alternative reporter plasmid for BMP15 and CD163 double-gene editing is constructed, and the editing condition of target sites of the BMP15 and CD163 genes of a cell genome can be converted into a detectable red fluorescence phenotype, so that the enrichment efficiency of anti-blue ear high-fecundity edited pig donor cells is improved, and the double-gene editing efficiency of the cells is effectively improved.

The invention discloses a biomarker for predicting skin melanoma metastasis and application thereof, and relates to the technical field of biological medicine. The biomarkers are mitochondrial dysfunction related proteins in macrophages in skin melanoma tissues and comprise GLUL and C3, the GLUL and the C3 are taken as the biomarkers, and the expression level of the GLUL and the C3 in the macrophages is detected through immunohistochemical staining, so that the metastasis prediction of the skin melanoma is realized. In the dyeing process, the macrophages are marked with CD163, then GLUL and C3 are specifically detected, an AEC red color developing agent is adopted, and a detection sample is a paraffin section of a primary focus and / or a metastatic focus of an SKCM patient. According to the invention, GLUL and C3 are determined as metastasis prediction biomarkers for the first time, and a specific immunohistochemical detection scheme is matched, so that double values of accurate prediction of metastasis risk and treatment guidance are realized, and a new technical path is provided for prognosis evaluation and individualized treatment of skin melanoma.

The present disclosure provides a monoclonal antibody that binds to porcine CD163, which is used for the treatment or prevention of porcine reproductive and respiratory syndrome (PRRS) virus infection of a pig, in particular to a monoclonal antibody that binds to porcine CD163, which is used for the treatment or prevention of porcine reproductive and respiratory syndrome (PRRS) virus infection of a pig. A preferred antibody comprises an antigen binding domain that binds porcine CD163, said antigen binding domain comprising at least one heavy chain variable region, said heavy chain variable region comprising three complementarity determining regions (CDRs), where said heavy chain variable region comprises a variable heavy chain (VH) CDR2, said variable heavy chain (VH) CDR2 comprising the amino acid sequence XYAD or XYAE or XYAN, where X may be any amino acid. Nucleic acid molecules, expression vectors and compositions are also provided.

The present disclosure relates to methods of treating certain cancers with a Siglec-9 extracellular domain (ECD) or a Siglec-9 ECD fusion molecule, including cancers expressing high levels of CD163 and / or Siglec-9 in tumor or immune cells, and methods of identifying cancers responsive to a Siglec-9 ECD or Siglec-9 ECD fusion molecule, for example by determining the level of CD163 and / or Siglec-9, and in some embodiments also CD68 and / or sialic acid expression in tumor or immune cells.

To provide methods of increasing resistance to porcine reproductive and respiratory syndrome virus in porcine animals, progeny, or cells.SOLUTION: A method of increasing resistance to porcine reproductive and respiratory syndrome virus comprises modifying at least one chromosomal sequence of a gene encoding a CD163 protein in a porcine animal, progeny, or cell, the modification being selected from the group consisting of a modification in exon 7 of the gene encoding the CD163 protein in which the modification comprises an in-frame deletion in exon 7, a modification in exon 8 of the gene encoding the CD163 protein, a modification in the intron adjacent to the exon 7 or exon 8 of the gene encoding the CD163 protein, and combinations thereof.SELECTED DRAWING: Figure 15

The invention relates to the technical field of animal gene engineering and biology, in particular to a pig MSTN, pAPN and CD163 gene synchronous editing embryo and a preparation method and application thereof. On the basis of a CRISPR / Cas9 technology, an MSTN exon 1, a CD163 exon 7 and a pAPN exon 2 are selected as targeting areas, efficient sgRNA is designed and screened, an integrated tandem editing vector containing a three-gene sgRNA expression cassette is constructed through homologous recombination, after pig embryo fibroblasts are electrically transfected, puromycin screening and sequencing identification are performed to obtain positive PEF cells, and the positive PEF cells are subjected to high-throughput screening and high-throughput screening. And performing somatic cell nuclear transfer to prepare and synchronously edit embryos. The synchronous editing efficiency of the three genes reaches 8.9%, the fusion rate of the edited embryos is larger than or equal to 70%, the cleavage rate is larger than or equal to 75%, the blastocyst rate is larger than or equal to 12%, the developmental ability is normal, synchronous improvement of genes related to growth performance and disease resistance is achieved, and the method has important industrial application value.

The invention discloses a bispecific antibody targeting heterogeneous macrophages, and relates to the field of biological medicines. According to the present invention, bevacizumab is adopted as a parent antibody skeleton, antibody arms targeting CD16 are arranged on the N end side, antibody arms targeting CD163 are located on the C end side, knob mutation (K322A and T366W) is introduced into the heavy chain CH3 structural domain targeting CD16, three complementary hole mutations (T366S, L368A and Y407V) are introduced into the heavy chain CH3 structural domain targeting CD163, and cysteine residues are inserted into the S354 site of the knob chain and the Y349 site of the hole chain; variable regions (V regions) targeting CD16 and CD163 are connected in series through a (G4S) 3 flexible linker and then fused with respective constant regions (C regions). The double-antibody architecture disclosed by the invention realizes collaborative optimization in structural stability, double-antigen binding specificity, macrophage targeting and functional activity, provides an efficient and safe novel candidate drug for targeted therapy of related diseases, and has important clinical transformation value and application prospect.

The invention discloses a prognostic marker for bladder cancer and application of the prognostic marker. The prognostic marker for the bladder cancer comprises CD14T, CD68S, CD163T and S100A12S; wherein the CD14T and the CD163T respectively refer to CD14 and CD163 expressed in a bladder cancer tumor region, and the CD68S and the S100A12S respectively refer to CD68 and S100A12 expressed in a bladder cancer matrix region. The bladder cancer prognosis marker provided by the invention can be used for constructing a bladder cancer prognosis prediction model, and multi-queue verification proves that the model has a strong prediction capability on the postoperative survival condition of a BLC patient and is beneficial to postoperative risk assessment and clinical decision of the BLC patient.

The present invention generally relates to genetically modified swine wherein at least one allele of a SIGLEC1 gene has been inactivated and / or at least one allele of a CD163 gene has been inactivated. Genetically modified swine having both alleles of the SIGLEC1 gene and / or both alleles CD163 gene inactivated are resistant to porcine reproductive and respiratory syndrome virus (PRRSV). Methods for producing such transgenic swine are also provided.

The invention belongs to the technical field of non-specific pathogen immune homeostasis evaluation, and provides a method for evaluating non-specific pathogen immune homeostasis by a CD163 gene edited pig, which comprises the following steps: peripheral blood sample collection, PBMC separation, sample induction, single cell library establishment and sequencing, cell annotation, standardization treatment, difference degree quantitative calculation and interval matching. According to the designed peripheral blood mononuclear cell immune homeostasis evaluation method based on single cell sequencing, directive information can be provided for animal experiments, the experiment cost is effectively reduced, the target performance is improved, and the repeatability of results is enhanced.

Provided herein are uses of antibodies that bind to CD163 expressed on a human myeloid cell in combination with checkpoint inhibitors. Among other things, these CD163 antibodies can be used with checkpoint inhibitors in methods of treatment of humans, such as methods of treating a cancer or methods of relieving T cell suppression.

The invention belongs to the field of biological medicine, and particularly relates to an antibody or an antigen binding fragment thereof and application. The antibody or the antigen binding fragment provided by the invention can specifically recognize the CD163 protein, for example, the binding EC50 value of the antibody or the antigen binding fragment provided by the invention and the hCD163-his recombinant protein can be as low as 1.712 ng / mL. In addition, the antibody or the antigen binding fragment can promote the M2 type macrophage to secrete TNF-alpha, reduce the expression of cell surface markers (such as CD16 and / or CD64) and promote the secretion of cytokines (such as IL-2 and / or IFN-gamma).