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

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...

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 used protein concentration techniques include dialysis bag concentration method, freeze-drying concentration method, blow-dry concentration method, ultrafiltration membrane concentration method, chemical precipitation method, etc. Ultrafiltration membrane concentration method and dialysis bag concentration method are relatively advanced, but the cost required It is relatively high, which is not conducive to popularization, and most other methods are likely to cause serum contamination and are difficult to use in cell culture. Based on the principle that freezing and thawing can precipitate and concentrate the chemical components of serum, we have found a practical method without affecting the activity of serum antigens. A feasible method for preparing autologous serum antigens, and then using these serums to sensitize DC, and cultivating specific antigen-sensitized DC-CIK cells for clinical use

Images