Preparation method for autologous-serum antigen-sensitized DC-CIK cells

A DC-CIK, autologous serum technology, applied in the fields of tumor immunotherapy and cell biology, can solve problems such as unfavorable promotion, high cost, serum pollution, etc., to avoid changes in antigen structure, low equipment requirements, and avoid resistance. Effect

Active Publication Date: 2014-08-13
SHENZHEN HORNETCORN BIOTECH
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since most tumors lack specific antigens, and tumors tend to mutate to resist the immune attack of a single antigen, the clinical application of specific antigen-sensitized DCs is greatly limited.
However, during the culture process of tumor cell lines, the surface antigens usually change, and the clinical effect of DC-CIK obtained after sensitized DC is not good, and the lysate of tumor specimens can only be used in patients whose specimens were collected before surgery. Most of the patients undergoing DC-CIK treatment are postoperative patients, and it is difficult to obtain fresh surgical specimens, which greatly limits the clinical effect of DC-CIK
However, due to individual differences in tumor patients, even the same type of tumors have very different tumor antigens, and artificially synthesized antigens cannot be widely used in the treatment of tumor patients.
[0009] Currently commonly us

Method used

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  • Preparation method for autologous-serum antigen-sensitized DC-CIK cells
  • Preparation method for autologous-serum antigen-sensitized DC-CIK cells
  • Preparation method for autologous-serum antigen-sensitized DC-CIK cells

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Example Embodiment

[0029] The following examples illustrate the present invention, but the present invention is not limited thereto. In the following examples, all experimental methods without specific conditions are carried out in accordance with conventional methods and manufacturer's operating instructions.

[0030] Routinely collect 50-100ml of peripheral blood of patients with advanced lung cancer, 5-15IU / ml of heparin sodium for anticoagulation, rotate at 800g, centrifuge for 10 minutes, collect the upper plasma part (about 1 / 3 of the liquid layer), and add the final concentration of 0.1-1.0 mg / ml leupeptin inhibitor, mix well, then store in a sterile centrifuge tube and freeze for 24-48h in an environment below -20°C (see figure 1 ), after taking it out, put it in a 0-4℃ environment for 10-16h (see figure 2 ); Inactivated at 56°C for 30 minutes, filtered with a 0.22μm microporous membrane to obtain autologous serum tumor antigens.

[0031] Density gradient centrifugation is used to separate P...

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Abstract

The invention discloses a preparation method for autologous-serum antigen-sensitized DC-CIK cells. The preparation method for the autologous-serum antigen-sensitized DC-CIK cells comprises the steps: A) preparation of an autologous-serum antigen; B) preparation of peripheral blood mononuclear cells; C) separation and culture of DC; D) induction amplification of CIK cells; and E) preparation of the DC-CIK cells through co-culture of DC cells and CIK cells. Compared with the DC-CIK cells prepared by employing a routine culture method, the DC-CIK cells cultured by employing the method are obviously increased in CIK proliferation activity and tumoricidal activity; the average proliferation multiple is 200-500 times and is 2-3 times of that of the DC-CIK cells cultured by employing the routine culture method; and the tumoricidal activity is 70-80% when an in-vitro experiment is performed, and is obviously higher than that of the DC-CIK cells cultured by employing the routine culture method. The method is simple in operation, easily controllable in conditions and relatively low in equipment requirements, and the cell proliferation efficiency of the obtained DC-CIK cells is relatively high.

Description

[0001] technical field [0002] The invention belongs to the fields of cell biology and tumor immunotherapy, and relates to a preparation method of DC-CIK sensitized by autologous tumor antigens and an application of anti-tumor adoptive immunotherapy. [0003] Background technique [0004] Malignant tumors have become the number one cause of death in China, seriously threatening the health and lives of the people. Tumor immunotherapy is the fourth major therapy for clinical tumor treatment, among which tumor adoptive immune cell therapy has been widely used in China. The most widely carried out is DC-CIK cell adoptive reinfusion therapy. [0005] DC cells are the most important antigen-presenting cells in the body. Through antigen uptake, processing and presentation, DC cells can effectively induce the activation of antigen-specific CD4 and CD8 T lymphocytes, thereby activating the body's adaptive immune response. In addition, DC can also activate NK cells, enhance their p...

Claims

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Application Information

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IPC IPC(8): C12N5/0783A61P35/00
Inventor 黄浩邹畅
Owner SHENZHEN HORNETCORN BIOTECH
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