Preparation of targeting CPI-444-loaded nanoparticles and investigation of its effects on T cell activity and anti-tumor response
DOI:10.3872/j.issn.1007-385x.2024.08.005
- VernacularTitle:靶向性CPI-444载药纳米微粒的制备及其对T细胞活性和抗肿瘤效应的影响
- Author:
CHEN Mingshui
1
;
LI Jieyu
1
;
WANG Ling
1
;
ZHOU Zhifeng
1
;
ZHANG Linteng
1
Author Information
1. Clinical Oncology School of Fujian Medical University, Fujian Cancer Hospital, Fuzhou 350014, Fujian, China
- Publication Type:Journal Article
- Keywords:
纳米微粒;腺苷受体A2A;过继细胞疗法
- From:
Chinese Journal of Cancer Biotherapy
2024;31(8):777-785
- CountryChina
- Language:Chinese
-
Abstract:
[摘 要] 目的:制备并表征包载CPI-444并偶联CD8抗体的纳米微粒(CNP/αCD8),探讨其对CD8+ T细胞活化、增殖和抗肿瘤作用的影响。方法: 采用复乳溶剂蒸发法和EDC/NHS法制备包载腺苷受体A2A(A2AR)特异性拮抗剂CPI-444(C)或香豆素6(C6)荧光素的纳米微粒并分别在其表面偶联CD8抗体,制得CNP/αCD8和C6NP/αCD8。扫描电镜和NanoPlus粒度测定仪表征纳米微粒形态和粒径,液相色谱与串联质谱联用(LC-MS/MS)法和离心法测定纳米微粒的载药量和药物释放情况,荧光显微镜和流式细胞仪检测CD8+ T细胞内化C6NP/αCD8的情况,流式细胞仪、ELISA和LDH法检测CNP/αCD8对CD8+ T细胞增殖、活化、细胞毒活性和杀瘤能力的影响。结果: CNP/αCD8纳米微粒为圆形、粒径约150 nm,能有效包载CPI-444和偶联CD8抗体,药物包封率和CD8抗体偶联效率分别约为60%和53.4%;CNP/αCD8纳米微粒具有良好稳定性,能被CD8+ T细胞内化,抑制A2AR分子表达。生物学功能实验显示,CNP/αCD8增强CD8+ T细胞的增殖能力、促进T细胞活化、分泌细胞因子及产生颗粒酶B和穿孔素,并增强CD8+ T细胞杀伤肿瘤细胞的能力。结论: CNP/αCD8纳米微粒能显著增强CD8+ T细胞免疫效应功能,其增强CD8+ T细胞功能可能是通过抑制A2AR分子的表达起作用。
- Full text:202409040901302933520240805.pdf