DC cell based on HBV1 antigen, targeting immune cell population, preparation method and applications thereof

A cell and antigen technology, applied to other methods of inserting foreign genetic materials, cells modified by introducing foreign genetic materials, animal cells, etc., can solve the problems of immune cell preparation methods that need to be improved, to solve the problem of cytotoxicity and efficiency High, good effect

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 HBV1 antigen, targeting immune cell population, preparation method and applications thereof
  • DC cell based on HBV1 antigen, targeting immune cell population, preparation method and applications thereof
  • DC cell based on HBV1 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] -80°C 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 micropipette guns (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); 75cm 2 Culture flask, 175cm 2 Culture flask, 1.8ml cryopreservation tube, 1.5ml EP tube, 50ml centrifuge tube, 15ml centrifuge tube, all from Coning Company, ...

Embodiment 2

[0132] Example 2: Clinical Treatment

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

[0134] 20 HBV1-positive liver cancer patients were divided into 2 groups (10 in group A and 10 in group B). For group A, after obtaining sufficient CTL cells on the 14th and 15th days according to the method in Example 1 , re-infused to the patient, the amount of cells re-infused each time is 1-2*10 9 ; Group B did not re-infuse cells (as a control). Blood collection and 2 consecutive re-infusions are a course of treatment. After 3 courses of treatment, the observation was carried out.

[0135] The results showed that there was a significant difference between group A and group B. All patients in group A felt less tired, increased appetite, less pain, and increased body weight after reinfusion, and 6 patients had decreased tumor indexes; The symptoms of group B were not improved. It can be clearly seen that ...

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Abstract

The invention discloses a DC cell based on HBV1 antigen, a targeting immune cell population, a preparation method and applications thereof. The preparation method of the 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 HBV1 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, the 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 present invention relates to the field of biotechnology, in particular, the present invention relates to DC cells, targeted immune cell populations, and preparation methods and uses thereof. Background technique [0002] Malignant tumors are the main types of lethal diseases in humans, and the mortality rate remains high. The latest statistics from the Ministry of Health show that malignant tumors are the first among the top ten causes of death among Chinese residents. In 2010, there were 2.68 million malignant tumor patients and 1.97 million deaths in China. At present, there are more than 6 million malignant tumor patients nationwide. The annual medical expenses for malignant tumor patients account for about 20% of the total health expenses, which is the largest medical burden in 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 tum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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