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CK19 antigen based DC cell and targeting immune cell population, and preparation method and application thereof

A cell and antigen technology, applied in the field of targeted immune cell populations and their preparation, DC cells, can solve the problems of immune cell preparation methods to be improved, and achieve the effect of solving cytotoxicity problems, high efficiency and high transfection efficiency

Inactive Publication Date: 2015-08-12
SHANGHAI TELEBIO BIOMEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the current preparation methods of immune cells for immune cell therapy still need to be improved

Method used

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  • CK19 antigen based DC cell and targeting immune cell population, and preparation method and application thereof
  • CK19 antigen based DC cell and targeting immune cell population, and preparation method and application thereof
  • CK19 antigen based DC cell and targeting immune cell population, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] 1. Materials and equipment

[0066] 1. Laboratory level: GLP

[0067] 2. Instruments and equipment:

[0068] Negative 80-degree ultra-low temperature refrigerator (Zhongke Meiling, DW-HL388); ordinary refrigerator (Haier, double door, -20°C and 4°C); mold incubator (Shanghai Boxun, MJX-160B-Z); constant temperature water bath ( SSW-420-2S); CO 2 Incubator (HF240); low-speed centrifuge (Shanghai Anting, TDL-40C); inverted biological microscope (Kexin, model: BLD-1); biological safety cabinet (Suzhou purification, double); Rand) 1-10ml; 1 set of micropipettes (Dalong), 4 pieces; programmed cooling box (Thermo); liquid nitrogen tank (Jinfeng, YDS-120-216); PCR instrument; gel electrophoresis imaging system ; Electrophoresis apparatus; Thermostat;

[0069] 3. Cell culture consumables:

[0070] Cell culture bag (Takara company); 75cm 2 Culture bottle, 175cm 2 Culture bottle, 1.8ml cryopreservation tube, 1.5ml EP tube, 50ml centrifuge tube, and 15ml centrifuge tube are...

Embodiment 2

[0132] Embodiment 2: clinical treatment

[0133] The CTL cells prepared in Example 1 were reinfused into the patient, and the reinfusion effect was observed, as follows:

[0134] Divide 20 CK19 positive nasopharyngeal carcinoma patients into 2 groups (10 patients in group A and 10 patients in group B), wherein, for group A, a sufficient amount of CTL was obtained on day 14 and day 15 according to the method in Example 1 After the cells, the patients were reinfused, and the amount of cells reinfused each time was 1~2*10 9 ; Group B did not reinfuse cells (as a control). One blood collection and two consecutive reinfusions constituted a course of treatment. After 3 courses of treatment, observation was carried out.

[0135] The results showed that there was a significant difference between group A and group B. After reinfusion, all patients in group A felt less tired, had increased appetite, lessened pain, and gained weight. Among them, 6 patients had decreased tumor indicato...

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Abstract

The invention discloses a DC cell and a targeting immune cell population, and a preparation method and application thereof. The preparation method of DC cells includes the steps of: conducting first induced differentiation culture on mononuclear cells in order to obtain immature DC cells; transfecting the immature DC cells by using ck19 antigen mRNA, so as to obtain transfected immature DC cells; and conducting second induced differentiation culture on the transfected immature DC cells, so as to obtain mature DC cells, wherein the mononuclear cells are isolated from mononuclear cells of peripheral blood of a sample. The method can prepare DC cells safely and efficiently, has a low cost and high efficiency, and can effectively solve the problem of cytotoxicity. The method can be effectively used for immune cell treatment or stimulation of lymphocytes derived from a same patient to produce a targeting cell population containing a large amount of CTL cells, and then the cell population can be used for tumor immunotherapy to reach better effect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, the invention relates to DC cells, targeted immune cell populations and their preparation methods and applications. Background technique [0002] Malignant tumors are the main type of fatal diseases in humans, and the mortality rate remains high. The latest statistics from the Ministry of Health show that malignant tumors rank first among the top ten causes of death among Chinese residents. In 2010, there were 2.68 million malignant tumor patients in China, and 1.97 million died. At present, there are more than 6 million malignant tumor patients in China. The annual medical expenses for malignant tumor patients account for about 20% of the total health expenses, which is the largest medical burden of the whole society. The current treatment methods are still very limited in achieving the goals of curing, prolonging the survival period of patients with malignant tumors, and improving ...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/87C12N5/0783A61K39/00A61K35/17A61P35/00
Inventor 杨光华李财新
Owner SHANGHAI TELEBIO BIOMEDICAL
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