Kim Yeongwon, Kim Sueon, Hong Cheol-Hwa, Hyun You-Seok, Baek In-Cheol, Kim Tai-Gyu
Department of Microbiology, College of Medicine, Catholic University of Korea, Seoul 06591, Korea.
Department of Biomedicine & Health Sciences, College of Medicine, Catholic University of Korea, Seoul 06591, Korea.
Vaccines (Basel). 2022 Nov 7;10(11):1877. doi: 10.3390/vaccines10111877.
Artificial antigen-presenting cells (aAPCs) that stably express particular HLA and co-stimulatory molecules by gene transfer have been developed to effectively stimulate T cells. To investigate whether cytochalsin-B-induced membrane vesicles derived from aAPCs (AP-CIMVs) have similar antigen-presenting functions as a cell-free system, T cell responses to different types of antigen presentation were measured using Jurkat reporter cells. First, the aggregation of AP-CIMV, which affects the measurement of function, was inhibited by nuclease treatment to produce uniform AP-CIMVs. The Green fluorescent protein (GFP) expression in Jurkat reporter cells was induced in a dose-dependent manner in groups stimulated with anti-CD3 antibody-coated AP-CIMVs and aAPCs, and anti-CD3/CD28 Dynabead. When Jurkat reporter cells expressing specific T cell receptors were stimulated by AP-CIMVs and aAPCs loaded with CMV pp65 peptide, AP-CIMVs showed similar stimulatory effects to that by aAPC. However, when these Jurkat reporter cells were stimulated by aAPCs endogenously expressing CMV pp65 antigen and their AP-CIMVs, the GFP expression rate by AP-CIMVs was 8.4%, which was significantly lower than 53.2% by aAPCs. Although this study showed a limited T-cell-stimulating effect of AP-CIMVs on endogenously processed antigen presentation, these results provide useful information for the development of improved cell-free systems for T cell stimulation in the future.
通过基因转移稳定表达特定人类白细胞抗原（HLA）和共刺激分子的人工抗原呈递细胞（aAPC）已被开发出来，用于有效刺激T细胞。为了研究细胞松弛素B诱导的源自aAPC的膜泡（AP-CIMV）作为无细胞系统是否具有类似的抗原呈递功能，使用Jurkat报告细胞测量了T细胞对不同类型抗原呈递的反应。首先，通过核酸酶处理抑制影响功能测量的AP-CIMV聚集，以产生均匀的AP-CIMV。在用抗CD3抗体包被的AP-CIMV、aAPC和抗CD3/CD28磁珠刺激的组中，Jurkat报告细胞中的绿色荧光蛋白（GFP）表达呈剂量依赖性诱导。当表达特异性T细胞受体的Jurkat报告细胞受到负载巨细胞病毒（CMV）pp65肽的AP-CIMV和aAPC刺激时，AP-CIMV显示出与aAPC相似的刺激作用。然而，当这些Jurkat报告细胞受到内源性表达CMV pp65抗原的aAPC及其AP-CIMV刺激时，AP-CIMV的GFP表达率为8.4%，显著低于aAPC的53.2%。尽管这项研究显示AP-CIMV对内源性处理的抗原呈递的T细胞刺激作用有限，但这些结果为未来开发改进的用于T细胞刺激的无细胞系统提供了有用信息。
