101 results about "CD163" patented technology

The invention provides a targeting sgRNA for pig CD163 gene editing, a modification vector, and a preparation method and application thereof, which relate to the technical field of gene engineering. The sgRNA and the gene modification vector provided by the invention have high specificity, and can be used for efficiently editing a pig CD163 gene on a cell level through a CRISPR/Cas9n system. According to the preparation method of a blue-eared disease resistant pig provided by the invention, a blue-eared disease virus receptor is damaged, the target gene CD163 is edited, no other exogenous gene is intruded, no non-specific editing is carried out on a non-CD163 gene area on a genome, a genetic background is clean and clear, and the work of later transgene safety evaluation is greatly reduced. The blue-eared disease resistant pig obtained by utilizing the invention can be ensured to survive normally, and resists the infection of a blue-eared disease virus, so that the economic loss caused by the blue-eared disease in pig husbandry is greatly reduced.

The invention discloses a method for editing a large white pig CD163 gene by using CRISPR/Cas9. The large white pig CD163 gene is edited by using the CRISPR/Cas9, the extracellular domain SRCR5 of a CD163 receptor is destroyed, the 7th exon DNA fragment of the CD163 gene is knocked out, and the nucleotide sequence of the 7th exon of the CD163 gene is represented by SEQ ID NO.1. The edited gene obtained by adopting the method can completely resist infection of PRRSV including a highly pathogenic strain HP-PRRSV, allows the cell surface CD163 receptor expression to be normal, and has normal other biological functions.

The invention relates to the field of immunology, gene therapy, and medicine. More specifically, the invention relates to the identification of a molecule capable of interacting with a soluble and/or cell-bound form of CD163 and, as a result of the interaction, an immune response is either instigated or suppressed in an organism. Furthermore, it relates to the preparation of a pharmaceutical composition including the molecule and/or antagonist I and/or agonist I thereof, and/or an isolated CD163 and/or an antagonist II or agonist II thereof, for the therapeutic or prophylactic treatment of an individual with an immune response disorder, e.g., inflammation, cancer, or infection.

This invention relates to targeted delivery of anti-viral compounds through protein bioconjugation. More particularly, it relates to an anti-viral compound conjugated to a protein, such as hemoglobin and to a method of treating a viral infection using said conjugate. The invention also provides a method of targeted drug delivery of an anti-viral nucleoside analogue to macrophages, cells comprising a hemoglobin receptor and to CD163 bearing cells.

The invention relates a method of quantifying CD64 and CD163 expression in leukocytes and, specifically to a kit for use with a flow cytometer including a suspension of quantitative fluorescent microbead standards, fluorescent labeled antibodies directed to CD64 and CD163, and analytical software. The software is used to take information on the microbead suspension and fluorescent labeled antibodies from a flow cytometer and analyse data, smooth curves, calculate new parameters, provide quality control measures and notify of expiration of the assay system.

The invention belongs to the technical field of preparing livestock molecular markers, and in particular relates to preparation and application of a molecular marker applied as porcine marker for assisted selection and relevant to porcine immunity. The molecular marker is obtained by cloning a porcine CD163 gene, and a mRNA sequence of the molecular marker is shown in a sequence table SEQ ID NO:1. A 2439bp locus of the sequence table SEQ ID NO:1 has an A-to-G base mutation; a 2454bp locus of the sequence has a G-to-C base mutation; a 2537bp locus of the sequence has a G-to-A base mutation; and a 2685bp locus of the sequence has an A-to-C base mutation. The invention also discloses a primer for amplifying partial coding regions of the CD163 gene and a method for detecting a 2537bp G-to-A molecular marker and a 2685bp A-to-C molecular marker. The invention provides two novel molecular markers for porcine marker-assisted selection.

The invention relates to a free receptor recombinant adenovirus group for expressing porcine reproductive and respiratory syndrome virus (PRRSV), and preparation and application thereof. The recombinant adenovirus group recombinant adenoviruses for expressing first four IgV structure fields of porcine sialic acid adhesin and porcine IgG Fc segment fusion proteins, and recombinant adenoviruses for expressing (5-9)th SRCR structure fields of virocapsid receptor CD163 and porcine IgG Fc segment fusion proteins. The recombinant adenovirus group is used for preparing a preparation for preventing and treating porcine reproductive and respiratory syndrome (PRRS).Compared with the existing vaccine, the recombinant adenovirus is replication-defective virus, can not be diffused and transmitted in the porcine body, and does not have the risk of toxicity diffusion and toxicity return; and the synergetic effect of the two expressed free receptors can obviously inhibiting PRRSV infection of different toxic strains, and can be developed as a novel preparation for preventing and controlling PRRS.

The invention belongs to the technical field of livestock gene engineering, and relates to construction and an application of a dual-luciferase reporter gene carrier based on a porcine CD163 gene. A key receptor gene, namely the porcine CD163 gene, of a PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) is amplified and sequenced to obtain a complete fragment of a 3' UTR (Untranslated Region) of the gene, candidate microRNAs (micro Ribonucleic Acids) and an action target of the 3' UTR of the porcine CD163 gene are predicted and regulated, and a sequence of the porcine CD163 gene containing the action target is amplified, and inserted into a dual-luciferase carrier to construct the dual-luciferase reporter gene carrier psi-CHECK2-CD163-3UTR of the 3' UTR of the porcine CD163 gene. The carrier can be used for detecting regulation activity of the candidate microRNAs on porcine CD163 gene expression, and screening the microRNAs inhibiting the porcine CD163 gene expression, and the like.

The invention discloses a porcine reproductive and respiratory syndrome resisting Marc-145 cell line as well as a preparation method and application thereof. The porcine reproductive and respiratory syndrome resisting Marc-145 cell line is obtained by knocking off CD163 extracellular domain SRCR5 of the Marc-145 cell line. The invention further provides gRNA (guide Ribonucleic Acid) for splicing of a pair of shearing Marc-145 cell CD163 receptors SRCR5 and sequences are shown as SEQ ID No. 2 and SEQ ID No. 3 respectively. After an the fifth extracellular fifth structure domain SRCR5 of the Marc-145 cell surface CD163 receptor on the surface of a Marc-145 cell is deleted through a CRISPR/CAS9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9) gene editing technology, the obtained porcine reproductive and respiratory syndrome resisting Marc-145 cell line can completely resist PRRSV (Porcine Reproductive and Respiratory Syndrome virus) infection and resist high-pathogenicity virulent strain HP-PRRSV. Moreover, the CD163 receptor on the surface of the porcine reproductive and respiratory syndrome resisting Marc-145 cell line surface CD163 receptor is normally expressed and other biological functions are normal. The porcine reproductive and respiratory syndrome resisting Marc-145 cell line disclosed by the invention has important meanings for inresearching PRRSV pathogenesis and mining novel disease-resisting or susceptible genes; a novel model is provided for PRRSV researches and the porcine reproductive and respiratory syndrome resisting Marc-145 cell line has important guidance meanings on in screening and breeding of PRRSV-resisting pigs.

The invention provides a pancreatic cancer detection reagent, a pancreatic cancer detection kit, a pancreatic cancer detection device and application. The detection reagent comprises antibodies of anytwo or more of the following molecular markers, wherein CD3, CD4, CD8 and CD68 belong to a group A, CD11b, CD14, CD33, CD45RO, CD57, CD66b, CD163, FoxP3, LAG3, PD1, PDL1, TIGIT, panCK and TIM3 belongto a group B, and at least one molecular marker is selected from the group B. Detecting pancreatic cancer through the antibodies of two or more of the molecular markers provided by the invention hasthe advantages of high detection sensitivity and excellent specificity. Thus, the molecular markers are suitable for a multiple immunohistochemistry method to detect tumor microenvironment of a pancreatic cancer patient and are very advantageous, and consequently, whether the patient has a longer lifetime can be accurately predicted relatively, namely, prognosis is better.

The invention relates to an esophageal squamous cell carcinoma microenvironmental cell marker molecular model and application thereof. The molecular model mainly comprises a-SMA+ (invasive frontier) fibroblasts, CD163+ (interstitial) macrophages, CD117+ (protogenic layer) mastocytes and CD117+ (invasion frontier) mastocytes. The molecular model can be used to predict the prognosis of patients withesophageal squamous carcinoma and group adjuvant treatment sensitivity groups. A kit mainly includes an anti-CD117 protein antibody, an anti-CD163 protein antibody and an anti-a-SMA protein antibody.According to the invention, the combination of three molecules combined with histopathological features predicts patients' prognosis, which has a higher predictive effect than single index detectionand clearly distinguishes patients with esophageal squamous carcinoma adjuvant treatment sensitivity; an immune tissue chemical hypersensitivity two-step method on which the molecular model is based is a mature and reliable method which can be widely used in primary hospitals; and compared with international TNM staging, the molecular model has a higher predictive effect, and is simpler, easier, more sensitive and more specific.

The invention provides a method for detecting CD163+/CD16+ cell population in peripheral blood mononuclear cells in a biological sample from a subject infected with HIV-1 which comprises contacting the biological sample with an anti-CD163 antibody, so that levels of CD163+/CD16+ peripheral blood mononuclear cells in the biological sample can be quantified. The method of the present invention is particularly useful for monitoring the course of HIV-1 infection and/or HIV Encephalopathy

The invention provides a reagent, a kit and a device for detecting tumor micro-environment of colorectal cancer as well as application of the reagent. The detection reagent comprises a POLE gene mutation detection reagent and an antibody of at least one molecular marker of at least one of the following combinations such as a combination 1: CD3, CD8, CD45RO, PD-1 and PD-L1 and a combination 2: CD4,FOXP3, CD68, CD163, PD-L1 and CD57. By utilizing the reagent, the kit and the device for detecting tumor micro-environment of colorectal cancer as well as the application of the reagent disclosed bythe invention, findings prove that POLE gene mutation is obviously related with high expression of a part of the molecular markers, and a relation between the POLE gene mutation and the tumor micro-environment of the colorectal cancer is determined, so that a multi-immunohistochemical method is conveniently and comprehensively utilized, and simultaneously several molecular markers are marked, andthe advantages that the sensitivity of detection on the tumor micro-environment of a pancreatic patient is high and the specificity is high are also utilized, so that whether a detected object has 5-year survival rate of higher probability can be relatively and accurately predicted.

The invention discloses a pig kidney cell line and application thereof. The preservation number of the pig kidney cell line is CCTCC (China Center for Type Culture Collection) C201324. A PK-15 cell line for stably expressing CD163 built by the pig kidney cell line can be applied to separation and cultivation of a porcine reproductive and respiratory syndrome (PRRSV) virus, and can also be applied to research of a PRRSV cell receptor. By building of a pig cell line susceptible to the PRRSV, an alternative cell line is provided for cultivation of the PRRSV; the range of a host cell for researching the PRRSV is enlarged; the pig kidney cell line has important significance on production of a PRRSV vaccine.

Provided is a double-gene knockout vector system, a method for preparing porcine fibroblasts with both CD163 gene and CD13 gene being knocked-out, prepared porcine fibroblasts, and a method for preparing a gene-edited pig with both CD163 gene and CD13 gene being knocked-out. The vector system of the present disclosure comprises a CD163 gene knockout vector and a CD13 gene knockout vector. The CD163 gene knockout vector comprises a gene editing vector backbone and a DNA fragment ligated to the gene editing vector backbone, with a nucleotide sequence of the DNA fragment being shown in any one of SEQ ID NOs: 1-3. The CD13 gene knockout vector comprises a gene editing vector backbone and a DNA fragment ligated to the gene editing vector backbone, a nucleotide sequence of the DNA fragment being shown in any one of SEQ ID NOs: 4-6.

The invention belongs to the technical field of malignant pleural effusion detection methods, and specifically relates to a method for identifying the malignant pleural effusion by using CD14+CD163+cells. The method comprises the following steps of extracting mononuclear cells, labeling antibodies, and performing immunophenotyping analysis by using a flow cytometry. Compared with the prior art, the method provided by the invention has the advantages that the method is simple and convenient and rapid, the sensitivity is high, the specificity is high, meanwhile, the interference of man-made subjective factors is reduced by using the method, the method is objective, judgment and identification can be made for the potential malignant pleural effusion patient as soon as possible, and convenience is provided for the subsequent in-time intervention treatment.

The invention relates to the technical fields of cytobiology, genetic engineering and veterinary biologicalogy, in particular to a construction method and application of a PK-15 stable cell strain. The construction method of the cell line for porcine reproductive and respiratory syndrome virus (PRRSV) breeding comprises the following steps that 1) a CD163 gene and an SBQ gene are constructed intoa lentiviral vector separately to obtain a recombinant lentiviral vector; 2) the recombinant lentiviral vector and a virus rescue helper plasmid are transferred into a mammalian cell, and the cell iscultured to obtain a recombinant virus; 3) a PK-15 cell is injected with the recombinant virus of step 2) to obtain the cell line. The invention further provides application of the cell. The obtainedcell has stable hereditary characters, is sensitive to the PRRSV, and has a very good application prospect.

The invention discloses an application of TREM-2 gene in identification of susceptibility of pigs to a reproductive and respiratory syndrome virus , and provides a group of TREM-2 expression level detection primer. The group of detection primers comprises an upstream primer TREM2-F represented by SEQ ID NO.1 and a downstream TREM2-R represented by SEQ ID NO.2. Researches show that the TREM-2 gene expression level in the pigs is related with PRRSV infection, and the TREM-2 can regulate the expression of CD163 in order to regulate duplication of the PRRSV; and the pigs can resist the PRRSV when the TREM-2 expression level is low, and the pigs are susceptible to the PRRSV when the TREM-2 expression level is high. The TREM-2 gene lays a solid foundation for identification of the susceptibility of pigs to the PRRSV and screening and cultivation of PRRSV-resistant pigs as a novel PRRSV susceptibility determination standard, and is of great significance to control the porcine reproductive and respiratory syndrome.

The present invention relates to a preparation method, an epitope identification and an application of a CD163 protein monoclonal antibody MAb. A functional structural domain SRCR5-6 with an importanteffect on PRRSV replication of CD163 is cloned into a pColdI vector, CD163 SRCR5-6 protein carrying His labels in a prokaryotic expression vector is constructed, the SRCR5-6 protein is purified to immunize Balb/c mice, spleen cells of mice and myeloma SP2/0 cells are taken to prepare hybridoma cells to obtain an anti-porcine CD163 protein monoclonal antibody, and besides, the antibody is subjected to the epitope identification.