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Virus specific target and application thereof in preparation of cellular immunotherapy preparation

A cellular immune and specific technology, applied in the field of virus-specific targets and its applications, can solve problems such as retention

Active Publication Date: 2015-06-10
河北博海生物工程开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The current clinical application of cellular immunotherapy technology basically stays at the level of LAK, NK, CIK, DC (clinical tumor biopsy tissue or tumor cell line extract), while the antigen-loaded and CAR-T technology by adenovirus transfection is due to the application of gene technology. , there are controversies in ethics and other aspects

Method used

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  • Virus specific target and application thereof in preparation of cellular immunotherapy preparation
  • Virus specific target and application thereof in preparation of cellular immunotherapy preparation
  • Virus specific target and application thereof in preparation of cellular immunotherapy preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 1. Specimen collection

[0020] The patient's blood count is required to be within the normal range (WBC: 4-10×10 9 , LYM%: 20%-40%), the fluctuation does not exceed 5%, the circulation volume of peripheral blood by machine sampling is 1000-4000ml, the number of mononuclear cells: more than 1×10 9 . Blood drawing method 50-100ml, the number of mononuclear cells: more than 1×10 6 .

[0021] The preferred anticoagulant is heparin, followed by sodium citrate, and EDTA is prohibited.

[0022] 2. Preparation before operation

[0023] Wipe the collection bag containing PBMC suspension from top to bottom with 75% alcohol, and then put it into the transfer window. Wipe it from top to bottom with a lint-free cloth of % alcohol, and then put it into the operating biological safety cabinet.

[0024] 3. Isolation of mononuclear cells

[0025] Transfer the PBMC cell suspension sample: Use sterilized scissors to cut the plastic tube, and carefully pour the PBMC cell suspension...

Embodiment 2

[0062] Example 2: Test method for detection of cell positive rate by target monoclonal antibody

[0063] Positive detection steps of indirect immunofluorescence on the surface of DC cells in peripheral blood of patients with hepatitis B

[0064] 1. Take the peripheral blood DC cells (1×10 7 / ml) 200 μl, divided into two tubes, one tube for each experimental control (100 μl / tube).

[0065] 2. Centrifuge and wash once with PBA,

[0066] 3. Add 20 μl each of FITC-CD86, PE-HLA-DR, and APC-CD11c to the experimental tube, and add 20 μl each of FITC-mouse IgG1, PE-mouse IgG1, and APC-mouse IgG1 to the control tube.

[0067] 4. Gently blow and beat to mix well. After each tube is fully mixed, incubate in the dark for 30 minutes.

[0068] 5. After washing with PBS, perform flow cytometry detection.

[0069] See the test results figure 1 As shown, the results show that co-culture of patient DC cells with hepatitis B DC targets in vitro can make the targets bind to the MHC in the DC...

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Abstract

The invention relates to the field of biotechnology, and discloses a virus specific target and an application thereof. The amino acid sequence of the hepatitis B virus specific target is FCWEWASAK. The invention further provides an application of the target in preparation of a preparation for treating hepatitis B related diseases. The target disclosed by the invention can realize in vitro effective amplification and activation of dendritic cells, the submitted T lymphocytes have specific killing effect to the virus, and the virus specific target is the cellular immunotherapy meeting the cellular immunology principle furthest, so the virus specific target has excellent application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to virus-specific targets and applications thereof. Background technique [0002] Viruses consist of a nucleic acid molecule (DNA or RNA) and protein or only protein (such as prions). Viruses are tiny and simple in structure. Viruses do not have a cellular structure and cannot replicate by themselves due to the absence of the basic systems necessary for metabolism. But when it contacts the host cell, it takes off the protein coat, and its nucleic acid invades the host cell, and with the help of the latter's replication system, it replicates new viruses according to the instructions of the viral gene. [0003] After virus infection, since the virus does not have the ability to reproduce itself, it must rely on the growth of human cells (ie target cells) to replicate. In other words, after virus infection, it must enter human cells to survive and replicate. After the human body is inf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C12N5/0784C12N5/0783A61K39/00A61P31/20
Inventor 李彬
Owner 河北博海生物工程开发有限公司
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