Vaccine for treating mRNA-DC lung cancer, enhanced preparation method of vaccine and CTL cell

A therapeutic vaccine and lung cancer stem cell technology, which is applied in the field of CTL cells, mRNA-DC lung cancer therapeutic vaccine and its preparation, can solve the problems of weak tumor killing ability, antigen cluster destruction, weak immune response, etc., and achieve shortened induction Time, comprehensive antigen information, and the effect of enhancing immune response

Inactive Publication Date: 2015-11-11
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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Problems solved by technology

However, there are deficiencies in this way of antigen stimulation. First, the antigen information carried by lung cancer cells is incomplete compared with the overall antigen information of lung cancer, and only a part of the antigen information can be transmitted to DC; second, the production of a certain protein is not enough. If, or if some broken proteins are produced, the antigenicity will be further reduced; thirdly, because the expansion...

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  • Vaccine for treating mRNA-DC lung cancer, enhanced preparation method of vaccine and CTL cell

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preparation example Construction

[0016] The example of the present invention proposes a kind of enhanced preparation method of mRNA-DC lung cancer therapeutic vaccine, comprises the following steps:

[0017] S10, obtaining the mRNA of lung cancer stem cells, as well as IL-2 and IFN-γ;

[0018] S20, transfecting the mRNA, IL-2 and IFN-γ of lung cancer stem cells into DC;

[0019] S30, culturing the DC transfected with the mRNA of lung cancer stem cells, IL-2 and IFN-γ in step S20 in vitro.

[0020] The preparation of the above-mentioned therapeutic vaccine of the present invention is based on DC cell immunity, and the maturation and antigen stimulation of DC are realized by mRNA transfection of lung cancer stem cells; after the test and investigation of DC, it can accept exogenous mRNA and Translate, because all organisms from the lowest virus to human codons have universality, after translation, the antigenic protein of lung cancer stem cells will be obtained, which can generate immune stimulation to DC, so ...

Embodiment 1

[0038] S11, obtaining the human lung adenocarcinoma cell line A549 (purchased from the tumor cell bank of the Chinese Academy of Medical Sciences);

[0039] S12. The lung adenocarcinoma cell line A549 obtained in step S11 was cultured in vitro in 3D. During the culture process, the A549 cells were cultured with RPMI1640 complete medium containing 10% fetal bovine serum at 37°C and 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Take the cell suspension in the logarithmic growth phase, and use 1×10 4 The concentration of 200 μl of each cell / well was inoculated in a 96-well plate covered with basal medium. After expiration, the basal medium was digested with dispase and the recovery solution was used to recover the cells.

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Abstract

The invention provides a vaccine for treating mRNA-DC lung cancer and an enhanced preparation method of the vaccine. The method includes the steps of obtaining mRNA, IL-2 and IFN-gamma of a lung cancer stem cell, and transfecting the mRNA, the IL-2 and the IFN-gamma into DC to be cultivated in vitro. According to the method process, based on the technological foundation that the DC can receive and translate the external mRNA and codon has universality, the DC is stimulated through the method that antigen proteins are expressed after the mRNA of the lung cancer stem cell is guided in and replace inactivated proteins for stimulation, and the carried antigen information is more comprehensive; the defect that due to inactivation, breakage and the like of the externally-stimulated antigen proteins, the antigen information is lost can be overcome; meanwhile, the IL-2 and the IFN-gamma are transfected into the DC, then the mature induction can be started in the immune response process, the induction time in an ordinary culture medium can be greatly shortened, the cell maturation efficiency is improved, and immune responses are enhanced. The invention further provides a cytotoxic T lymphocyte obtained by co-culturing the vaccine for treating mRNA-DC lung cancer and a T lymphocyte.

Description

technical field [0001] The invention belongs to the technical field of cellular immunotherapy, and in particular relates to an mRNA-DC lung cancer therapeutic vaccine, a preparation method thereof and CTL cells. Background technique [0002] The adoptive immunotherapy for lung cancer is to induce, activate and amplify the precursor cells of self or allogeneic anti-tumor effector cells in vitro with activators such as IL-2, anti-CD3 monoclonal antibody, and specific polypeptides, and then transfect Infuse it to tumor patients, so as to improve the anti-tumor immunity of patients, so as to achieve the purpose of treatment. Currently, the adoptive immune cells most used in clinical practice are mainly DC (dendritic cell, dendritic cell) and CIK (Cytokine-Induced Killer, cytokine-induced killer cell). [0003] Among them, DC is isolated from bone marrow tissue and blood, has strong ability to capture antigen, present antigen and secrete ability, and can induce a lasting and pow...

Claims

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Application Information

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IPC IPC(8): A61K35/12A61K39/00A61K39/39A61P35/00C12N5/0783
Inventor 曾宪卓鲁菲
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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