CAJ | 학술논문

目的：比较并建立 T 细胞体外活化的条件，为后续功能及机制相关实验奠定基础。方法以 Jur-kat 细胞为模型，选择 T 细胞早期活化的指标 CD69分子作为评价指标，通过比较不同刺激条件下 CD69分子的表达量的差别，来判定合适的体外刺激条件，并通过增殖实验、抑制性信号表达量以及 PBMC 细胞进行验证。结果多克隆刺激剂 PMA 和 ionomycin 在剂量分别为20和500 ng/ml 时， Jurkat 细胞活化效率较高；单克隆刺激剂 anti-CD3包被和 anti-CD28游离，且剂量在5μg/ml 和1．5μg/ml 时 Jurkat 细胞活化效率较高；作用48 h 检测多克隆刺激剂活化效率略高于单克隆刺激剂，细胞发生了增殖、抑制性分子 CTLA-4的表达增加。结论建立了 Jurkat 及 PBMC 细胞的稳定的体外刺激条件，为后续研究 T 细胞的功能等奠定了实验基础。
목적：비교병건립 T 세포체외활화적조건，위후속공능급궤제상관실험전정기출。방법이 Jur-kat 세포위모형，선택 T 세포조기활화적지표 CD69분자작위평개지표，통과비교불동자격조건하 CD69분자적표체량적차별，래판정합괄적체외자격조건，병통과증식실험、억제성신호표체량이급 PBMC 세포진행험증。결과다극륭자격제 PMA 화 ionomycin 재제량분별위20화500 ng/ml 시， Jurkat 세포활화효솔교고；단극륭자격제 anti-CD3포피화 anti-CD28유리，차제량재5μg/ml 화1．5μg/ml 시 Jurkat 세포활화효솔교고；작용48 h 검측다극륭자격제활화효솔략고우단극륭자격제，세포발생료증식、억제성분자 CTLA-4적표체증가。결론건립료 Jurkat 급 PBMC 세포적은정적체외자격조건，위후속연구 T 세포적공능등전정료실험기출。
Objective To compare the efficacy of different stimuli in T cell activation in vitro in order to facilitate T cell function studies.Methods Jurkat cells were chosen as a cell model, and CD69 as the early activation marker.A suitable stimulus condition was determined in vitro by comparing the expression difference of CD69 in different stimulus settings.The optimal condition was then confirmed by cell proliferation, cytotoxic T lymphocyte antigen 4 (CTLA-4) expression and PBMC activation.Results Jurkat cells were conspicuously activated when stimulated with 20 ng/mL PMA and 500 ng/mL ionomycin.They were greatly activated when treated with coated anti -hu-man CD3 at 5 μg/mL and free anti-human CD28 at 1.5 μg/mL.Forty-eight hours after stimulation, the activation efficacy with PMA and ionomycin was higher than that with anti -CD3 and -CD28, as evidenced by increased cell proliferation and over -expression of CTLA-4.Conclusion A stable T cell activation condition was established in vitro , which provides a solid foundation for T cell func -tion studies.