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miRNA-155 and application of its inhibitor in DC-CIK cell culture
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A technology of mirna-155, 1. mirna-155, applied in the field of immunization, can solve the problems of unsatisfactory effect and poor efficacy of adoptive immunization
Active Publication Date: 2017-05-31
朗姿赛尔生物科技(广州)有限公司
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[0005] Although a large number of studies have shown that DCs, DC tumor vaccines, and CIK cells have significant anti-tumor effects, clinical applications have found that DCs and CIK cells alone are not very effective in treating tumors. Tumor cellsresist these immune effector cells and lead to adoptive immunity. Poor efficacy
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Embodiment 1
[0024] Example 1: The effect of miRNA-155 on DC-CIK cell co-culture
[0029] 2. The tumor cells are leukemia cells, including K562 / A02, THP-1 and HL-60 cells.
[0030] 2. Experimental method
[0031] 1. The separation and culture of effector cells DC and CIK
[0032] (1) Separation of mononuclear cells: Collect 20mL of peripheral blood from healthy volunteers, dilute 1:1 with pre-cooled PBS, slowly add the upper layer of lymphocyte separation solution, 650g, centrifuge for 20min at 4°C, collect white cell layer, and separate mononuclear cells , Resuspend the cells in 1640 medium and place them at 37℃, 5% CO 2 Incubate for 2h in t...
Embodiment 2
[0082] Example 2: Screening of miRNA-155 inhibitors (Pseudostellaria japonicus B, C)
[0083] 1. Experimental materials
[0084] Pseudoginsenopsis cyclic peptide B was purchased from Shanghai Yuanye Biotechnology Co., Ltd.; Pseudoginsenopsis cyclic peptide C was purchased from Kunming Institute of Botany, Chinese Academy of Sciences. Other materials are the same as in Example 1.
[0085] 2. Experimental method
[0086] 1. The separation and culture of effector cells DC and CIK
[0087] (1) Separation of mononuclear cells: Collect 20mL of peripheral blood from healthy volunteers, dilute 1:1 with pre-cooled PBS, slowly add the upper layer of lymphocyte separation solution, 650g, centrifuge for 20min at 4°C, collect white cell layer, and separate mononuclear cells , Resuspend the cells in 1640 medium and place them at 37℃, 5% CO 2 Incubate for 2h in the incubator.
[0088] (2) Cultivation of DC cells: After mononuclear cells are cultured for 2 hours, adherent cells are collected and induc...
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Abstract
The invention discloses a miRNA-155 and an application of its inhibitor in a DC-CIK cell culture; the miRNA-155 can be used as a drug target and applied to improve the killability of a CIK cell co-cultured and enhanced by a DC cell to a cancer cell. The applicant study finds that the DC-CIK cell under down-regulated expression of miRNA-155 has higher killability than the DC-CIK cell under normal expression of miRNA-155, and the killability of the DC cell under down-regulated expression of miRNA-155 is basically consistent with the killability of the DC cell under the normal expression of miRNA-155, there is no significant difference. It indicates that the miRNA-155 down-regulation cannot directly improve the killability of the DC cell to leukemia cell, but the down-regulated DC cell of the miRNA-155 can significantly enhance the killability of the cancer cell through co-culture. The applicant further finds that radix pseudostellariae cyclic peptide B and radix pseudostellariae cyclic peptide C are effective inhibitors of miRNA-155.
Description
Technical field [0001] The invention belongs to the field of immunity, relates to the cultivation of DC-CIK cells, and specifically relates to the application of miRNA-155 and its inhibitors in the cultivation of DC-CIK cells. Background technique [0002] Biological immunotherapy is to supplement, induce or activate the inherent biological response regulation system in the body from outside, activate and mobilize biologically active cells and cytokines with cytotoxic activity to adjust various immune-killing biological responses. At present, there are more research applications in immunotherapy: lymphokine-activated killer cells, tumor infiltrating lymphocytes, cytokine-induced killer (CIK) cells, dendritic cells (DC), and co-cultured immune (DC-CIK) cells , Natural killer cell type lymphocytes, among which DC and CIK cells are two important parts of tumor immunotherapy. The immune response induced by their interaction is the central link of the immunosuppressive effect [Wang Zh...
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