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DC cell based on LMP-1 antigen, targeting immune cell population, preparation method and applications thereof

An LMP-1, cell technology, applied to other methods of inserting foreign genetic material, cells modified by introducing foreign genetic material, animal cells, etc., can solve the problem of immune cell preparation methods need to be improved, and achieve the problem of cytotoxicity , high efficiency, 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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  • DC cell based on LMP-1 antigen, targeting immune cell population, preparation method and applications thereof
  • DC cell based on LMP-1 antigen, targeting immune cell population, preparation method and applications thereof
  • DC cell based on LMP-1 antigen, targeting immune cell population, preparation method and applications 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] 20 LMP-1 positive herpes virus patients were divided into 2 groups (10 people in group A and 10 people in group B), wherein, for group A, according to the method in Example 1, a sufficient amount was obtained on the 14th day and the 15th day After CTL cells, reinfuse the patient, and the amount of cells reinfused each time is 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, increased appetite, less pain, and increased weight. Among them, 6 patients had less symptoms of herpes virus infection;...

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PUM

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Abstract

The invention discloses a DC cell based on LMP-1 antigen, a targeting immune cell population, a preparation method and applications thereof. The preparation method of DC cell comprises the following steps: subjecting mononuclear cells to first induced differentiation culture to obtain immature DC cells; transfecting the immature DC cells by LMP-1 antigen mRNA to obtain transfected immature DC cells; subjecting the transfected immature DC cell to second induced differentiation culture to obtain mature DC cells; wherein the mononuclear cells are separated from peripheral blood mononuclear cells of a sample. Through the provided preparation method, DC cells can be obtained efficiently and safely. Moreover, the provided preparation method has the advantages of low cost and high efficiency, and can effectively solve the problem of cell toxicity. The DC cells can be applied to cell immunotherapy or used to trigger the lymph cells from the same patient to generate targeting immune cell population rich in CTL cells so as to achieve a better effect in tumor immunotherapy.

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