三阴性乳腺癌细胞裂解物通过PD1/PDL1相互作用抑制免疫细胞活性

Triple negative breast cancer cell lysate inhibits immune cell activity via PD1/PDL1 interaction

目的:研究三阴性乳腺癌肿瘤细胞裂解物(TCL)通过PD1/PDL1相互作用对T淋巴细胞系H9凋亡、活化及细胞因子分泌情况的影响。方法:Western blot检测MDA-MD-231、MCF-10A细胞中的PDL1和H9、PBMC中的PD1表达;反复冻融法制备231-TCL,将不同剂量231-TCL作用H9细胞,于不同时间点观察231-TCL诱导细胞凋亡的剂量与时间;适量的231-TCL与PD1/PDL1抑制剂共同作用H9细胞,观察H9细胞凋亡、CD69表达及IL-2、IL-4、TNF-β的分泌。结果:MDA-MD-231、MCF-10A细胞中均有PDL1表达,H9、PBMC中表达PD1;当231-TCL∶H9为2∶1,且作用时间为48 h时,231-TCL可诱导H9细胞凋亡;231-TCL和PD1/PDL1抑制剂联合作用于H9细胞后,H9细胞凋亡减少,CD69表达增加,IL-2、TNF-β分泌增加,IL-4分泌量不变。结论:三阴性乳腺癌TCL通过PD1/PDL1相互作用诱导H9细胞凋亡,抑制其活化及IL-2、TNF-β等细胞因子的分泌。

Objective:To study the effect of triple-negative breast cancer tumor cell lysate(TCL)on the apoptosis,activation and cytokine secretion of T lymphocyte line H9 through the interaction of PD1/PDL1.Methods:The expression of PDL1 in MDA-MD-231 and MCF-10A and PD1 in H9 and PBMC were detected by Western blot;231-TCL was prepared by repeated freezing and thawing method.Different doses of 231-TCL to H9 cells were given at different time points,then observed the well dose and time of 231-TCL induced apoptosis;Use 231-TCL and PD1/PDL1 inhibitor were used to act on H9 cells to observe H9 apoptosis,CD69 expression,secretion of IL-2,IL-4,and TNF-β.Results:PDL1 was expressed in MDA-MD-231,MCF-10A cells,and PD1 was expressed in H9cells,PBMC;when the ratio of 231-TCL∶H9 was 2∶1 and the action time was 48 h,231-TCL could induce H9 cells apoptosis well.After 231-TCL and PD1/PDL1 inhibitor are used in combination with H9 cells,H9 cells apoptosis decreases,CD69 expression increases,IL-2,TNF-βsecretion increases,and IL-4 secretion remains unchanged.Conclusion:Triple-negative breast cancer TCL induces apoptosis of H9 cells and inhibits their activation and secretion of cytokines such as IL-2 and TNF-βthrough PD1/PDL1 interaction.