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DC-CIK细胞通过抗Tim-3抗体和抗程序性细胞死亡蛋白-1抗体对肺腺癌的增强抑制作用及可能机制

Enhanced Inhibitory Effect of DC-CIK Cells on Lung Adenocarcinoma via Anti-Tim-3 Antibody and Antiprogrammed Cell Death-1 Antibody and Possible Mechanism.

作者信息

Zhou Liang, Chen Qijiu, Chen Hui, Wang Li, Zhang Jianyong

机构信息

The Second Department of Respiratory and Critical Care Medicine, Affiliated Hospital of Zunyi Medical University, Zunyi 563000, Guizhou, China.

Department of Respiratory and Critical Care Medicine, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, Guangdong, China.

出版信息

Evid Based Complement Alternat Med. 2022 Jul 31;2022:4097576. doi: 10.1155/2022/4097576. eCollection 2022.

DOI:10.1155/2022/4097576
PMID:35958924
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9357700/
Abstract

OBJECTIVE: To investigate the effect and mechanism of blocking the signaling pathways of the T-cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) and programmed death protein 1 (PD-1) in dendritic cell-cytokine induced killer (DC-CIK) cells on human lung adenocarcinoma A549 cells. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated and induced into mature DC-CIK cells by cytokines . After blocking the Tim-3 and PD-1 signaling transduction pathways with anti-Tim-3 and anti-PD-1 antibodies, DC-CIK cells were coincubated with A549 cells. The killing effect of DC-CIK cells against A549 cells was measured by a CCK-8 assay. The impact of DC-CIK cells on the invasion and migration ability of A549 cells was detected by the Transwell test. The apoptosis rate of DC-CIK cells and the ratio of CD4, CD8, and DC-CIK cell subsets were determined by flow cytometry. The cell proliferation of DC-CIK was detected by the CCK-8 assay. RESULTS: The antibodies of anti-Tim-3 antibody and anti-PD-1 could block Tim-3 and PD-1 DC-CIK cells and could significantly increase the killing effect of DC-CIK cells on A549 cells. The number of A549 cells under the microporous membrane of the Transwell chamber was reduced considerably in invasion and migration tests. Anti-Tim-3 and anti-PD-1 antibodies significantly reduced apoptosis of DC-CIK cells. No significant differences were observed in the ratios of CD4 and CD8 DC-CIK cell subsets or the proliferation capacity of DC-CIK cells in each group. CONCLUSION: Blocking the Tim-3 and PD-1 signaling pathways of DC-CIK cells with antibodies can enhance the killing ability of DC-CIK cells in A549 cells and significantly suppress the invasion and migration ability of A549 cells. The potential mechanism may be related to reduced apoptosis of DC-CIK cells.

摘要

目的：探讨阻断树突状细胞-细胞因子诱导的杀伤细胞（DC-CIK）中T细胞免疫球蛋白黏蛋白结构域3（Tim-3）和程序性死亡蛋白1（PD-1）信号通路对人肺腺癌A549细胞的作用及机制。 方法：分离外周血单个核细胞（PBMC），并用细胞因子诱导其成为成熟的DC-CIK细胞。用抗Tim-3和抗PD-1抗体阻断Tim-3和PD-1信号转导通路后，将DC-CIK细胞与A549细胞共孵育。采用CCK-8法检测DC-CIK细胞对A549细胞的杀伤作用。通过Transwell试验检测DC-CIK细胞对A549细胞侵袭和迁移能力的影响。采用流式细胞术检测DC-CIK细胞的凋亡率以及CD4、CD8和DC-CIK细胞亚群的比例。采用CCK-8法检测DC-CIK细胞的增殖情况。 结果：抗Tim-3抗体和抗PD-1抗体可阻断DC-CIK细胞中的Tim-3和PD-1，并可显著提高DC-CIK细胞对A549细胞的杀伤作用。在侵袭和迁移试验中，Transwell小室微孔膜下的A549细胞数量显著减少。抗Tim-3和抗PD-1抗体显著降低了DC-CIK细胞的凋亡。各组DC-CIK细胞亚群的CD4和CD8比例或DC-CIK细胞的增殖能力均未观察到显著差异。 结论：用抗体阻断DC-CIK细胞的Tim-3和PD-1信号通路可增强DC-CIK细胞对A549细胞的杀伤能力，并显著抑制A549细胞的侵袭和迁移能力。潜在机制可能与DC-CIK细胞凋亡减少有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/9d0413253081/ECAM2022-4097576.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/022fd3c7fe46/ECAM2022-4097576.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/2f2e59179370/ECAM2022-4097576.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/1903539788be/ECAM2022-4097576.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/21f88e3ad418/ECAM2022-4097576.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/bf116f799ad7/ECAM2022-4097576.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/c265957c467f/ECAM2022-4097576.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/d4b57310d75f/ECAM2022-4097576.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/9d0413253081/ECAM2022-4097576.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/022fd3c7fe46/ECAM2022-4097576.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/2f2e59179370/ECAM2022-4097576.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/1903539788be/ECAM2022-4097576.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/21f88e3ad418/ECAM2022-4097576.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/bf116f799ad7/ECAM2022-4097576.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/c265957c467f/ECAM2022-4097576.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/d4b57310d75f/ECAM2022-4097576.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/72c6/9357700/9d0413253081/ECAM2022-4097576.008.jpg

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