Preparation method and application of a new tumor dendritic cell therapeutic vaccine

A technology of dendritic cells and tumor cells, which is applied in the field of preparation of novel high-efficiency dendritic cell tumor therapeutic vaccines, can solve the problems of limited curative effect, difficult quality control, complicated operation, etc.

Active Publication Date: 2011-11-30
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology

[0008] However, there are obvious deficiencies in the above four tumor vaccines at present. Many tumor antigens are not clear and cannot induce tumor-specific CTLs. The use of dendritic cell tumor vaccines and genetically modified tumor cell tumor vaccines also has complex in vitro operations, Factors such as difficult quality control and limited curative effect

Method used

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  • Preparation method and application of a new tumor dendritic cell therapeutic vaccine
  • Preparation method and application of a new tumor dendritic cell therapeutic vaccine
  • Preparation method and application of a new tumor dendritic cell therapeutic vaccine

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preparation example Construction

[0061] The present invention also provides a method for preparing the tumor antigen-containing supernatant, which mainly includes the steps of:

[0062] (a) Inducing and culturing the tumor cells with the chemotherapeutic drug 5-FU at 37±3°C for 0.5-48 hours to obtain a culture medium; and

[0063] (b) centrifuging the culture solution in step (a), so as to obtain the culture supernatant.

[0064] In another preferred example, in step (a), at a concentration of 1.0×10 -6 M to 1.0×10 -2 M (preferably 0.5-1.5×10 -3 M) In the presence of 5-FU, induction culture was performed.

[0065] In another preferred embodiment, in step (b), centrifugation is performed at 5000-20000 rpm.

[0066] In another aspect, the present invention also provides a method for dendritic cells sensitized by the above supernatant, comprising the following steps:

[0067] The dendritic cells are sensitized with the tumor antigen-containing supernatant described in the first aspect of the present inventi...

Embodiment 1

[0087] Preparation of New High Efficiency Tumor Vaccine

[0088] Human tumor cells (human colon cancer cell SW480) cultured in RPMI-1640 complete medium (containing 10% fetal bovine serum) were collected, and the concentration was adjusted to 1×10 6 / ml, adding a final concentration of 1.0×10 -3 M of 5-Fu, then incubated in a 37°C incubator for 12 hours, centrifuged at 10,000rpm for 10 minutes, and the supernatant was collected as a tumor antigen.

[0089] The tumor antigen induced by chemotherapeutic drugs obtained as above was prepared according to the tumor antigen 50 μg / 10 6 On the 6th day of DC sensitization for 48 hours, this new and highly effective vaccine for tumor treatment and prevention can be obtained. observations under a microscope, such as figure 1 shown.

Embodiment 2

[0091] Chemotherapy drug-induced tumor antigens include immune stimulatory molecules Hsp70, HMGB1

[0092] Human colon cancer cell SW480 cultured in RPMI-1640 complete medium (containing 10% fetal bovine serum) was collected, and the concentration was adjusted to 1×10 6 / ml, 1ml / well was placed in a 24-well plate, and 0.5×10 -3 M, 1.0×10 -3 M, 1.5×10 -3 For 5-Fu of M, cells and supernatant were collected 0, 4, 12, and 24 hours after the action, and heat shock protein 70 (Hsp70), high-speed swimming group protein B1 (HMGB1) Flisa kit (RD company), The concentration of immunostimulatory molecules in the cell supernatant was detected.

[0093] The result is as figure 2 , image 3shown. The results showed that the chemotherapeutic drug-induced tumor antigens contained the immunostimulatory molecules Hsp70 and HMGB1.

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Abstract

The invention provides a preparation method and application of a new therapeutic vaccine for dendritic tumor cells. The therapeutic tumor vaccine is chemotherapeutic drug induced tumor antigen, wherein, the dendritic cells are loaded and activated. The chemotherapeutic medicament induced tumor antigen contains a plurality of immunostimulation molecules as well as tumor-associated and specific antigen, which can remarkably carry out chemotaxis and activation on immunocytes such as the dendritic cells, T-cells (thymus-dependent lymphocytes) and the like, stimulate the dendritic cells to be mature and express a plurality of cell factors and chemotactic factors, effectively induce immune response reaction with antigenic specificity and non-specificity and strengthen body immune function. The therapeutic vaccine provided by the invention can be used for preventing and treating tumors and has the characteristics of simple preparation process, low cost, strong specificity, obvious curative effect and the like.

Description

technical field [0001] The invention relates to the fields of biology and medicine, and more specifically relates to a preparation method of a novel high-efficiency dendritic cell tumor therapeutic vaccine and the application of the vaccine in the treatment and prevention of various tumors. Background technique [0002] Active immunotherapy of tumors is an effective means to treat and prevent tumors or infectious diseases. Since humans used vaccinia to prevent smallpox, humans have made great progress in the prevention and control of infectious diseases. At present, it has basically Infectious diseases such as smallpox and polio have been eradicated, but there is still a lack of effective means for the treatment of other major diseases such as the prevention and treatment of tumors and other diseases. [0003] Tumor immunotherapy is a new treatment model after surgery, radiotherapy, and chemotherapy. The central link of tumor immunotherapy is to induce the body to produce sp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/39C12N5/0784A61P35/00
Inventor 万涛昂冰黄晓辉曹雪涛
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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