A kind of fusion cell and its preparation method and its application as tumor vaccine

Abstract

The invention discloses a fusion cell and its preparation method and its application as a tumor vaccine. The method comprises the following steps: (1) preparing tumor cells modified by NDV-Ulster virus strain; (2) preparing fusion cells using phosphoantigen or bisphosphine A new type of antigen-presenting cell γδT-APC stimulated and activated by salt drugs; (3) preparation of fusion cells of γδT-APC and NDV-Ulser modified tumor cells; (4) purification of fusion cells; (5) cultivation of fusion cells; (6) Preparation of fusion cell products. The method of the present invention can obtain more novel antigen-presenting cells γδT-APC, and the fusion rate of γδT-APC and the modified tumor cells is high, and the tumor antigen titer of the fused cells obtained in the present invention is more complete, the immunogenicity is stronger, High tumor antigen loading rate.

Description

technical field [0001] The present invention relates to a fusion cell and its preparation method and the application of the fusion cell as a tumor vaccine, in particular to a fusion preparation of tumor cells modified by novel antigen-presenting cell γδT-APC and Newcastle disease strain NDV-Ulser The cell method also relates to the use of the resulting fused cell as a tumor vaccine. Background technique [0002] Dendritic cells (hereinafter referred to as Dendritic Cells, DCs) as professional antigen-presenting cells play a vital role in inducing anti-pathogenic microorganisms and anti-tumor immune responses. DC-based anti-cancer vaccines have also received increasing attention in clinical trials. In order to obtain tumor-specific vaccines, researchers have adopted various strategies to develop tumor vaccines based on DC cells to stimulate anti-tumor-specific immune responses. DC vaccines used in clinical trials mainly include antigen-loaded DC cell vaccines, gene-modified...

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Problems solved by technology

[0004] However, DC-tumor fusion vaccines also have their great limitations. Although DC cells can be induced by monocytes in peripheral blood, the ability of monocyte-derived DC (Mo-DC) to proliferate in vitro and different DC induction conditions Due to the restriction of mononuclear DC (Mo-DC), the number of mononuclear-derived DC (Mo-DC) is very limited. Generally, 100ml of peripheral blood is isolated from PBMCs and can obtain up to 1×10 after induction of adherence. 7 Moreover, the homogeneity of these Mo-DC cells is very poor, and mature and immature DC are often intertwined together. The purity of immature DC cells can generally only reach about 40-50%, and the fusion efficiency of unmodified DC-tumor About 30%, according to the ratio of DC cells: tumor cells = 1:10, the fusion rate is only 20-30%, and the expression of antigens related to fusion cells can only reach about 30-50%, so clinical trials In order to achieve the ideal therapeutic effect, the DC-tumor fusion cell vaccine prepared in the method must meet the following conditions: first, both antigen-presenting cells and tumor cells used to prepare fusion cells must have strong immunogenicity; It is necessary to improve the fusion efficiency of antigen-presenting cells and tumor cells. Only a sufficient number of fused cells can better exert an effective immune response; the third is to obtain a sufficient number of fused cells, first of all, there must be a sufficient number and a uniform phenotype antigen presenting cells

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