Application of PLPP1 in preparation of T cell immune tumor related medicament
A PLPP1, 1.PLPP1 technology is applied in the direction of anti-tumor drugs, vaccines, intestinal cancer vaccines, etc., and can solve the problems affecting the response of T cells to blocking therapy and the effect of tumor therapy
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[0032] The present application will be further explained below in conjunction with the accompanying drawings and embodiments. Before introducing the specific embodiments, the experimental background of some biological materials, experimental reagents, experimental equipment, etc. involved in the following embodiments is briefly introduced as follows.
[0033] biomaterials:
[0034] Peripheral blood from patients with esophageal cancer, lung cancer or colon cancer, as well as adjacent normal tissues and tumor tissues, were all obtained from relevant departments of the First Affiliated Hospital of Zhengzhou University after approval by the Ethics Committee; peripheral blood from healthy donors came from Zhengzhou City, Henan Province Red Cross Blood Center; lung cancer cell lines H460 and A549, from the Institute of Cells, Chinese Academy of Sciences; SPF grade CD45.1 C57BL / 6N wild-type female mice, from Peking University School of Basic Medicine; SPF grade CD45.2 OT-1 mice, An...
Embodiment 1
[0045] The inventor believes that the microenvironment of solid tumors is closely related to the function of T cells, and the formation of the microenvironment is the result of the joint action of T cells and tumor tissues. In order to explore and determine the tumor-related T cells in the tumor microenvironment The inventors conducted a series of experiments on the key influencing factors of metabolism, which are specifically introduced as follows.
[0046] (1) Differences in related expression molecules
[0047] In order to determine the actual difference in molecular expression, the inventors took samples of peripheral blood and tumor tissue from lung cancer patients, processed them into small fragments of 2-4 mm, treated them with three enzymes (enzyme H, enzyme R, enzyme A) and performed tissue digestion Finally, human peripheral blood mononuclear cells (PBMC) were extracted by density gradient centrifugation, CD8+ T cells were further separated by magnetic separation usi...
Embodiment 2
[0059] On the basis of Example 1, the inventors further used genetic engineering technology to knock down PLPP1 to study and analyze the influence of PLPP1 on the metabolism and function of activated CD8+ T cells. The specific research process is as follows.
[0060] (1) Flow cytometric analysis of functional molecular differences in CD8+ T cells with low and high expression of PLPP1
[0061] In Example 1, the existing database statistics have preliminarily shown that the expression of PLPP1 in CD8+ T cells is related to its function and metabolism. In order to further verify this association, the inventors used flow cytometry to detect PLPP1 in the tumor microenvironment of patients with lung cancer high and PLPP1 low Related factors in CD8+ T cells (for details, please refer to the existing technology for operation). The result is as figure 2 shown. Analysis can be seen: with PLPP1 low CD8+ T cells compared to PLPP1 high CD8+ T cells can secrete higher levels of kil...
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