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DC cell based on HBV-HCV 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, high production 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 HBV-HCV antigen, targeting immune cell population, preparation method and applications thereof
  • DC cell based on HBV-HCV antigen, targeting immune cell population, preparation method and applications thereof
  • DC cell based on HBV-HCV 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. Instrument and equipment:

[0068] Minus 80 degrees ultra-low temperature refrigerator (Zhongke Meiling, DW-HL388); ordinary refrigerator (Haier, double door, -20℃ and 4℃); 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); electric pipette (German general RAND) 1-10ml; Micro pipette (Dalong) 1 set, 4 pieces; Program cooling box (Thermo); Liquid nitrogen tank (Jinfeng, YDS-120-216); PCR machine; Gel electrophoresis imaging system ; Electrophoresis instrument; Thermostat;

[0069] 3. Cell culture consumables:

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

Embodiment 2

[0132] Example 2: Clinical treatment

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

[0134] Twenty HBV-HCV-positive liver cancer patients were divided into 2 groups (10 in group A and 10 in group B). For group A, sufficient CTLs were obtained on the 14th and 15th day according to the method in Example 1. After the cells are transfused back to the patient, the amount of cells transfused each time is 1~2*10 9 ; Group B does not return cells (as a control). Blood is collected once and infused twice continuously as a course of treatment. After 3 courses of treatment, observe.

[0135] The results showed that there was a clear difference between group A and group B. All patients in group A felt less physical exhaustion, increased appetite, reduced pain, and weight gain after reinfusion. Among them, 6 patients had tumor indicators decreased; and The symptoms of group B did not improve. It can be clear...

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Abstract

The invention discloses a DC cell based on HBV-HCV 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 HBV-HCV 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. Specifically, 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 type of human fatal diseases, 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, 2.68 million patients suffered from malignant tumors in China and 1.97 million died; there are currently more than 6 million patients with malignant tumors nationwide. The annual medical expenses for patients with malignant tumors 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 tumors...

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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